Sample records for alcohol oxidase aod

  1. Preparation of convection interaction media isobutyl disc monolithic column and its application to purification of secondary alcohol dehydrogenase and alcohol oxidase.

    PubMed

    Isobe, Kimiyasu; Kawakami, Yoshimitsu

    2007-03-09

    A convection interaction media (trade name CIM, BIA Separation, Ljubljana, Slovenia) isobutyl monolithic disc was prepared by incubating a CIM epoxy monolithic disc with isobutylamine, and it was then applied to the purification of secondary alcohol dehydrogenase (S-ADH) and primary alcohol oxidase (P-AOD). Both enzymes were adsorbed on this column and eluted with high purity. Thus, S-ADH was purified to an electrophoretically homogeneous state by four column chromatographies using CIM DEAE-8 and CIM C4-8 tube monolithic columns, blue-Sepharose column and CIM isobutyl disc monolithic column. P-AOD was also purified to an electrophoretically homogeneous state by three column chromatographies of CIM DEAE-8 tube, CIM C4-8 tube and CIM isobutyl disc columns.

  2. Alternative Oxidase Transcription Factors AOD2 and AOD5 of Neurospora crassa Control the Expression of Genes Involved in Energy Production and Metabolism.

    PubMed

    Qi, Zhigang; Smith, Kristina M; Bredeweg, Erin L; Bosnjak, Natasa; Freitag, Michael; Nargang, Frank E

    2017-02-09

    In Neurospora crassa , blocking the function of the standard mitochondrial electron transport chain results in the induction of an alternative oxidase (AOX). AOX transfers electrons directly from ubiquinol to molecular oxygen. AOX serves as a model of retrograde regulation since it is encoded by a nuclear gene that is regulated in response to signals from mitochondria. The N. crassa transcription factors AOD2 and AOD5 are necessary for the expression of the AOX gene. To gain insight into the mechanism by which these factors function, and to determine if they have roles in the expression of additional genes in N. crassa , we constructed strains expressing only tagged versions of the proteins. Cell fractionation experiments showed that both proteins are localized to the nucleus under both AOX inducing and noninducing conditions. Furthermore, chromatin immunoprecipitation and high throughput sequencing (ChIP-seq) analysis revealed that the proteins are bound to the promoter region of the AOX gene under both conditions. ChIP-seq also showed that the transcription factors bind to the upstream regions of a number of genes that are involved in energy production and metabolism. Dependence on AOD2 and AOD5 for the expression of several of these genes was verified by quantitative PCR. The majority of ChIP-seq peaks observed were enriched for both AOD2 and AOD5. However, we also observed occasional sites where one factor appeared to bind preferentially. The most striking of these was a conserved sequence that bound large amounts of AOD2 but little AOD5. This sequence was found within a 310 bp repeat unit that occurs at several locations in the genome. Copyright © 2017 Qi et al.

  3. Amperometric sensor for ethanol based on one-step electropolymerization of thionine-carbon nanofiber nanocomposite containing alcohol oxidase.

    PubMed

    Wu, Lina; McIntosh, Mike; Zhang, Xueji; Ju, Huangxian

    2007-12-15

    Thionine had strong interaction with carbon nanofiber (CNF) and was used in the non-covalent functionalization of carbon nanofiber for the preparation of stable thionine-CNF nanocomposite with good dispersion. With a simple one-step electrochemical polymerization of thionine-CNF nanocomposite and alcohol oxidase (AOD), a stable poly(thionine)-CNF/AOD biocomposite film was formed on electrode surface. Based on the excellent catalytic activity of the biocomposite film toward reduction of dissolved oxygen, a sensitive ethanol biosensor was proposed. The ethanol biosensor could monitor ethanol ranging from 2.0 to 252 microM with a detection limit of 1.7 microM. It displayed a rapid response, an expanded linear response range as well as excellent reproducibility and stability. The combination of catalytic activity of CNF and the promising feature of the biocomposite with one-step non-manual technique favored the sensitive determination of ethanol with improved analytical capabilities.

  4. Immobilization of Pichia pastoris cells containing alcohol oxidase activity

    PubMed Central

    Maleknia, S; Ahmadi, H; Norouzian, D

    2011-01-01

    Background and Objectives The attempts were made to describe the development of a whole cell immobilization of P. pastoris by entrapping the cells in polyacrylamide gel beads. The alcohol oxidase activity of the whole cell Pichia pastoris was evaluated in comparison with yeast biomass production. Materials and Methods Methylotrophic yeast P. pastoris was obtained from Collection of Standard Microorganisms, Department of Bacterial Vaccines, Pasteur Institute of Iran (CSMPI). Stock culture was maintained on YPD agar plates. Alcohol oxidase was strongly induced by addition of 0.5% methanol as the carbon source. The cells were harvested by centrifugation then permeabilized. Finally the cells were immobilized in polyacrylamide gel beads. The activity of alcohol oxidase was determined by method of Tane et al. Results At the end of the logarithmic phase of cell culture, the alcohol oxidase activity of the whole cell P. Pastoris reached the highest level. In comparison, the alcohol oxidase activity was measured in an immobilized P. pastoris when entrapped in polyacrylamide gel beads. The alcohol oxidase activity of cells was induced by addition of 0.5% methanol as the carbon source. The cells were permeabilized by cetyltrimethylammonium bromide (CTAB) and immobilized. CTAB was also found to increase the gel permeability. Alcohol oxidase activity of immobilized cells was then quantitated by ABTS/POD spectrophotometric method at OD 420. There was a 14% increase in alcohol oxidase activity in immobilized cells as compared with free cells. By addition of 2-butanol as a substrate, the relative activity of alcohol oxidase was significantly higher as compared with other substrates added to the reaction media. Conclusion Immobilization of cells could eliminate lengthy and expensive procedures of enzyme separation and purification, protect and stabilize enzyme activity, and perform easy separation of the enzyme from the reaction media. PMID:22530090

  5. Crystal Structure of Alcohol Oxidase from Pichia pastoris

    PubMed Central

    Valerius, Oliver; Feussner, Ivo; Ficner, Ralf

    2016-01-01

    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. PMID:26905908

  6. Cloning and Analysis of the Alternative Oxidase Gene of Neurospora Crassa

    PubMed Central

    Li, Q.; Ritzel, R. G.; McLean, LLT.; McIntosh, L.; Ko, T.; Bertrand, H.; Nargang, F. E.

    1996-01-01

    Mitochondria of Neurospora crassa contain a cyanide-resistant alternative respiratory pathway in addition to the cytochrome pathway. The alternative oxidase is present only when electron flow through the cytochrome chain is restricted. Both genomic and cDNA copies for the alternative oxidase gene have been isolated and analyzed. The sequence of the predicted protein is homologous to that of other species. The mRNA for the alternative oxidase is scarce in wild-type cultures grown under normal conditions, but it is abundant in cultures grown in the presence of chloramphenicol, an inhibitor of mitochondrial protein synthesis, or in mutants deficient in mitochondrial cytochromes. Thus, induction of alternative oxidase appears to be at the transcriptional level. Restriction fragment length polymorphism mapping of the isolated gene demonstrated that it is located in a position corresponding to the aod-1 locus. Sequence analysis of mutant aod-1 alleles reveals mutations affecting the coding sequence of the alternative oxidase. The level of aod-1 mRNA in an aod-2 mutant strain that had been grown in the presence of chloramphenicol was reduced several fold relative to wild-type, supporting the hypothesis that the product of aod-2 is required for optimal expression of aod-1. PMID:8770590

  7. AOD treatment agencies: does religious affiliation influence service delivery?

    PubMed

    McIlwraith, Fairlie; Kinner, Stuart A; Najman, Jake M

    2011-11-01

    Religious organisations have been involved in delivering alcohol and other drug (AOD) services since Australian colonial times and are a familiar presence in the AOD sector. However, there is concern in some sectors that AOD services delivered by religious organisations might be influenced by religious ideology, at the expense of evidence-based service provision. A national, cross-sectional survey of non-government AOD agencies was undertaken using a mailed questionnaire. All non-government AOD agencies in Australia, providing at least one face-to-face specialist AOD service, were invited to participate. Agency goals and activities were assessed using the Drug and Alcohol Program Treatment Inventory, which has eight distinct treatment orientations: 12-step, therapeutic community, cognitive behavioural therapy, psychodynamic, family, rehabilitation, dual diagnosis and medical. There was a high degree of uniformity in treatment orientations with religiously affiliated agencies having similar goals and activities to non-religiously affiliated agencies. Cognitive behavioural therapy was most commonly provided and 12-step the least provided. Religiously affiliated agencies were significantly more likely to favour the 12-step orientation in both goals and activities. Concerns that the religious affiliation of non-government organisations might influence AOD service delivery in Australia appear to be overstated. Factors contributing to the observed uniformity of care may include a more strategic, federal approach; and an increasing emphasis on best practice within the sector. The lack of discernable differentiation between religiously affiliated and non-religiously affiliated non-government organisations may also be attributable to changes in the way services are delivered by many religious organisations. © 2011 Australasian Professional Society on Alcohol and other Drugs.

  8. Spectral and catalytic properties of aryl-alcohol oxidase, a fungal flavoenzyme acting on polyunsaturated alcohols

    PubMed Central

    2005-01-01

    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 oxidative dehydrogenation of a wide range of unsaturated primary alcohols with hydrogen peroxide production. This differentiates the enzyme from VAO (vanillyl-alcohol oxidase), which is specific for phenolic compounds. Moreover, AAO is optimally active in the pH range of 5–6, whereas VAO has an optimum at pH 10. Kinetic studies showed that AAO is most active with p-anisyl alcohol and 2,4-hexadien-1-ol. AAO converts m- and p-chlorinated benzyl alcohols at a similar rate as it does benzyl alcohol, but introduction of a p-methoxy substituent in benzyl alcohol increases the reaction rate approx. 5-fold. AAO also exhibits low activity on aromatic aldehydes. 19F NMR analysis showed that fluorinated benzaldehydes are converted into the corresponding benzoic acids. Inhibition studies revealed that the AAO active site can bind a wide range of aromatic ligands, chavicol (4-allylphenol) and p-anisic (4-methoxybenzoic) acid being the best competitive inhibitors. Uncompetitive inhibition was observed with 4-methoxybenzylamine. The properties described above render AAO a unique oxidase. The possible mechanism of AAO binding and oxidation of substrates is discussed in the light of the results of the inhibition and kinetic studies. PMID:15813702

  9. Weekday AOD smaller than weekend AOD in eastern China on the basis of the MODIS AOD product

    NASA Astrophysics Data System (ADS)

    Song, Jingjing; Xia, Xiangao; Zhang, Xiaoling; Che, Huizheng; Li, Xiaojing

    2018-05-01

    A weekly cycle of surface particulate matter (PM) characterized by smaller values during weekends and larger values during weekdays was reported in eastern China. Whether column-integrated aerosol optical depth (AOD) showed similar weekly cycling as that of PM was debated. The weekly variation of AOD in eastern China was further studied by using the latest MODIS aerosol product (collection 6) with a fine spatial resolution (0.1°) from 2002 to 2015. We used three statistical methods to determine whether the weekly cycle of AOD was significant. AOD during weekdays (Wednesday to Friday) was lower than that during weekends. The maximum and minimum AOD was generally observed on Monday and Wednesday, respectively. This weekly pattern of AOD was in good agreement with previous results based on satellite aerosol products with a coarse spatial resolution, but it was in contrast to that of PM. Further analysis of the AOD weekly variability in 19 provincial cities suggested that AOD during weekdays was smaller than that during weekends in urban regions. Potential causes for the different weekly cycle of PM and AOD in eastern China were discussed.

  10. Stepwise Hydrogen Atom and Proton Transfers in Dioxygen Reduction by Aryl-Alcohol Oxidase.

    PubMed

    Carro, Juan; Ferreira, Patricia; Martínez, Angel T; Gadda, Giovanni

    2018-03-20

    The mechanism of dioxygen reduction by the flavoenzyme aryl-alcohol oxidase was investigated with kinetic isotope, viscosity, and pL (pH/pD) effects in rapid kinetics experiments by stopped-flow spectrophotometry of the oxidative half-reaction of the enzyme. Double mixing of the enzyme in a stopped-flow spectrophotometer with [α- 2 H 2 ]- p-methoxybenzyl alcohol and oxygen at varying aging times established a slow rate constant of 0.0023 s -1 for the wash-out of the D atom from the N5 atom of the reduced flavin. Thus, the deuterated substrate could be used to probe the cleavage of the N-H bond of the reduced flavin in the oxidative half-reaction. A significant and pH-independent substrate kinetic isotope effect (KIE) of 1.5 between pH 5.0 and 8.0 demonstrated that H transfer is partially limiting the oxidative half-reaction of the enzyme; a negligible solvent KIE of 1.0 between pD 5.0 and 8.0 proved a fast H + transfer reaction that does not contribute to determining the flavin oxidation rates. Thus, a mechanism for dioxygen reduction in which the H atom originating from the reduced flavin and a H + from a solvent exchangeable site are transferred in separate kinetic steps is proposed. The spectroscopic and kinetic data presented also showed a lack of stabilization of transient flavin intermediates. The substantial differences in the mechanistic details of O 2 reduction by aryl-alcohol oxidase with respect to other alcohol oxidases like choline oxidase, pyranose 2-oxidase, and glucose oxidase further demonstrate the high level of versatility of the flavin cofactor in flavoenzymes.

  11. Comparison of Government and Non-Government Alcohol and Other Drug (AOD) Treatment Service Delivery for the Lesbian, Gay, Bisexual, and Transgender (LGBT) Community.

    PubMed

    Mullens, Amy B; Fischer, Jane; Stewart, Mary; Kenny, Kathryn; Garvey, Shane; Debattista, Joseph

    2017-07-03

    Lesbian, gay, bisexual, and transgender (LGBT) populations are more likely to misuse alcohol and other drugs (AOD), compared to the general population. However, LGBT engagement with AOD treatment is often precluded by insensitivity and misunderstanding of LGBT issues. These treatment barriers may be a consequence of either worker attitudes, organizational factors or a combination of both. Few studies have compared service context as an impediment to AOD treatment. This pilot study sought to examine and compare staff attitudes, knowledge and awareness of LGBT issues in two state-wide AOD services within Australia. One organization was a government service, whilst the other was faith based. A cross-sectional study of a convenience sample (N = 130) of workers employed in a state-wide government AOD service (n = 65), and a state-wide non-government service (n = 65) was conducted. Participants self-completed a questionnaire comprising tools previously used to assess staff attitudes, knowledge and awareness of LGBT issues. Few significant differences in attitudes and awareness of LGBT issues between government and non-government respondents were found. Nearly all respondents were supportive of LGBT persons irrespective of organizational context, with a small number of negative views. Although most respondents demonstrated awareness of organizational policies and practices relating to LGBT clients, many were "unsure" or "neutral" of what these might be. It is confirming that the majority of staff report supportive attitudes towards LGBT clients. Findings suggest that organizations need to continue to take leadership to strengthen organizational training and capacity to deliver LGBT friendly AOD treatment practices.

  12. Quantitative assessment of AOD from 17 CMIP5 models based on satellite-derived AOD over India

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

    Misra, Amit; Kanawade, Vijay P.; Tripathi, Sachchida Nand

    Aerosol optical depth (AOD) values from 17 CMIP5 models are compared with Moderate Resolution Imaging Spectroradiometer (MODIS) and Multiangle Imaging Spectroradiometer (MISR) derived AODs over India. The objective is to identify the cases of successful AOD simulation by CMIP5 models, considering satellite-derived AOD as a benchmark. Six years of AOD data (2000–2005) from MISR and MODIS are processed to create quality-assured gridded AOD maps over India, which are compared with corresponding maps of 17 CMIP5 models at the same grid resolution. Intercomparison of model and satellite data shows that model-AOD is better correlated with MISR-derived AOD than MODIS. The correlation between model-AOD andmore » MISR-AOD is used to segregate the models into three categories identifying their performance in simulating the AOD over India. Maps of correlation between model-AOD and MISR-/MODIS-AOD are generated to provide quantitative information about the intercomparison. The two sets of data are examined for different seasons and years to examine the seasonal and interannual variation in the correlation coefficients. In conclusion, latitudinal and longitudinal variations in AOD as simulated by models are also examined and compared with corresponding variations observed by satellites.« less

  13. Quantitative assessment of AOD from 17 CMIP5 models based on satellite-derived AOD over India

    DOE PAGES

    Misra, Amit; Kanawade, Vijay P.; Tripathi, Sachchida Nand

    2016-08-03

    Aerosol optical depth (AOD) values from 17 CMIP5 models are compared with Moderate Resolution Imaging Spectroradiometer (MODIS) and Multiangle Imaging Spectroradiometer (MISR) derived AODs over India. The objective is to identify the cases of successful AOD simulation by CMIP5 models, considering satellite-derived AOD as a benchmark. Six years of AOD data (2000–2005) from MISR and MODIS are processed to create quality-assured gridded AOD maps over India, which are compared with corresponding maps of 17 CMIP5 models at the same grid resolution. Intercomparison of model and satellite data shows that model-AOD is better correlated with MISR-derived AOD than MODIS. The correlation between model-AOD andmore » MISR-AOD is used to segregate the models into three categories identifying their performance in simulating the AOD over India. Maps of correlation between model-AOD and MISR-/MODIS-AOD are generated to provide quantitative information about the intercomparison. The two sets of data are examined for different seasons and years to examine the seasonal and interannual variation in the correlation coefficients. In conclusion, latitudinal and longitudinal variations in AOD as simulated by models are also examined and compared with corresponding variations observed by satellites.« less

  14. Alcohol biosensing by polyamidoamine (PAMAM)/cysteamine/alcohol oxidase-modified gold electrode.

    PubMed

    Akin, Mehriban; Yuksel, Merve; Geyik, Caner; Odaci, Dilek; Bluma, Arne; Höpfner, Tim; Beutel, Sascha; Scheper, Thomas; Timur, Suna

    2010-01-01

    A highly stable and sensitive amperometric alcohol biosensor was developed by immobilizing alcohol oxidase (AOX) through Polyamidoamine (PAMAM) dendrimers on a cysteamine-modified gold electrode surface. Ethanol determination is based on the consumption of dissolved oxygen content due to the enzymatic reaction. The decrease in oxygen level was monitored at -0.7 V vs. Ag/AgCl and correlated with ethanol concentration. Optimization of variables affecting the system was performed. The optimized ethanol biosensor showed a wide linearity from 0.025 to 1.0 mM with 100 s response time and detection limit of (LOD) 0.016 mM. In the characterization studies, besides linearity some parameters such as operational and storage stability, reproducibility, repeatability, and substrate specificity were studied in detail. Stability studies showed a good preservation of the bioanalytical properties of the sensor, 67% of its initial sensitivity was kept after 1 month storage at 4 degrees C. The analytical characteristics of the system were also evaluated for alcohol determination in flow injection analysis (FIA) mode. Finally, proposed biosensor was applied for ethanol analysis in various alcoholic beverage as well as offline monitoring of alcohol production through the yeast cultivation. Copyright 2010 American Institute of Chemical Engineers

  15. Costs of alcohol and drug-involved crime.

    PubMed

    Miller, Ted R; Levy, David T; Cohen, Mark A; Cox, Kenya L C

    2006-12-01

    A large proportion of violent and property crimes involve alcohol or other drugs (AOD). AOD use only causes some of these crimes. This paper estimates the costs of AOD-involved and AOD-attributable crimes. Crime counts are from government statistics adjusted for underreporting. The AOD-involved portion of crime costs is estimated from inmate surveys on alcohol and illicit drug use at the time of the crime. The costs and AOD-attributable portion of AOD-involved crimes come from published studies. They include tangible medical, mental health, property loss, future earnings, public services, adjudication, and sanctioning costs, as well as the value of pain and suffering. An estimated 5.4 million violent crimes and 8 million property crimes involved AOD use in 1999. Those AOD-involved crimes cost society over 6.5 billion dollars in medical and mental health care and almost 65 billion dollars in other tangible expenses (in 1999 dollars). If the value of pain, suffering, and lost quality of life is added, AOD-involved crime costs totaled 205 billion dollars. Violent crimes accounted for more than 85% of the costs. Roughly estimated, crimes attributable to alcohol cost 84 billion dollars, more than 2 times the 38 billion dollars attributable to drugs. Although American media--news and entertainment--dwell on the links between drugs and crime, alcohol-attributable crime costs are double drug-attributable ones. Effective efforts to reduce the abuse of alcohol and illicit drugs should reduce costs associated with crime.

  16. Regio- and Stereospecific Conversion of 4-Alkylphenols by the Covalent Flavoprotein Vanillyl-Alcohol Oxidase

    PubMed Central

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

    1998-01-01

    The regio- and stereospecific conversion of prochiral 4-alkylphenols by the covalent flavoprotein vanillyl-alcohol oxidase was investigated. The enzyme was active, with 4-alkylphenols bearing aliphatic side chains of up to seven carbon atoms. Optimal catalytic efficiency occurred with 4-ethylphenol and 4-n-propylphenols. These short-chain 4-alkylphenols are stereoselectively hydroxylated to the corresponding (R)-1-(4′-hydroxyphenyl)alcohols (F. P. Drijfhout, M. W. Fraaije, H. Jongejan, W. J. H. van Berkel, and M. C. R. Franssen, Biotechnol. Bioeng. 59:171–177, 1998). (S)-1-(4′-Hydroxyphenyl)ethanol was found to be a far better substrate than (R)-1-(4′-hydroxyphenyl)ethanol, explaining why during the enzymatic conversion of 4-ethylphenol nearly no 4-hydroxyacetophenone is formed. Medium-chain 4-alkylphenols were exclusively converted by vanillyl-alcohol oxidase to the corresponding 1-(4′-hydroxyphenyl)alkenes. The relative cis-trans stereochemistry of these reactions was strongly dependent on the nature of the alkyl side chain. The enzymatic conversion of 4-sec-butylphenol resulted in two (4′-hydroxyphenyl)-sec-butene isomers with identical masses but different fragmentation patterns. We conclude that the water accessibility of the enzyme active site and the orientation of the hydrophobic alkyl side chain of the substrate are of major importance in determining the regiospecific and stereochemical outcome of vanillyl-alcohol oxidase-mediated conversions of 4-alkylphenols. PMID:9791114

  17. A rapid and sensitive alcohol oxidase/catalase conductometric biosensor for alcohol determination.

    PubMed

    Hnaien, M; Lagarde, F; Jaffrezic-Renault, N

    2010-04-15

    A new conductometric biosensor has been developed for the determination of short chain primary aliphatic alcohols. The biosensor assembly was prepared through immobilization of alcohol oxidase from Hansenula sp. and bovine liver catalase in a photoreticulated poly(vinyl alcohol) membrane at the surface of interdigitated microelectrodes. The local conductivity increased rapidly after alcohol addition, reaching steady-state within 10 min. The sensitivity was maximal for methanol (0.394+/-0.004 microS microM(-1), n=5) and decreased by increasing the alcohol chain length. The response was linear up to 75 microM for methanol, 70 microM for ethanol and 65 microM for 1-propanol and limits of detection were 0.5 microM, 1 microM and 3 microM, respectively (S/N=3). No significant loss of the enzyme activities was observed after 3 months of storage at 4 degrees C in a 20mM phosphate buffer solution pH 7.2 (two or three measurements per week). After 4 months, 95% of the initial signal still remained. The biosensor response to ethanol was not significantly affected by acetic, lactic, ascorbic, malic, oxalic, citric, tartaric acids or glucose. The bi-enzymatic sensor was successfully applied to the determination of ethanol in different alcoholic beverages. (c) 2009 Elsevier B.V. All rights reserved.

  18. In vivo relationship between monoamine oxidase type B and alcohol dehydrogenase: effects of ethanol and phenylethylamine

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

    Aliyu, S.U.; Upahi, L.

    The role of acute ethanol and phenylethylamine on the brain and platelet monoamine oxidase activities, hepatic cytosolic alcohol dehydrogenase, redox state and motor behavior were studied in male rats. Ethanol on its own decreased the redox couple ratio, as well as, alcohol dehydrogenase activity in the liver while at the same time it increased brain and platelet monoamine oxidase activity due to lower Km with no change in Vmax. The elevation in both brain and platelet MAO activity was associated with ethanol-induced hypomotility in the rats. Co-administration of phenylethylamine and ethanol to the animals, caused antagonism of the ethanol-induced effectsmore » described above. The effects of phenylethylamine alone, on the above mentioned biochemical and behavioral indices, are more complex. Phenylethylamine on its own, like ethanol, caused reduction of the cytosolic redox, ratio and elevation of monoamine oxidase activity in the brain and platelets. However, in contrast to ethanol, this monoamine produced hypermotility and activation of the hepatic cytosolic alcohol dehydrogenase activity in the animals.« less

  19. Attitudes, Skills and Knowledge Change in Child and Adolescent Mental Health Workers Following AOD Screening and Brief Intervention Training

    ERIC Educational Resources Information Center

    Christie, Grant; Black, Stella; Dunbar, Lucy; Pulford, Justin; Wheeler, Amanda

    2013-01-01

    Adolescent mental health workers are generally poor at identifying and treating co-existing alcohol and other drug (AOD) disorder. This study aimed to evaluate the utility and acceptability of an AOD screening and brief intervention (BI) training package delivered to child and adolescent mental health workers and its impact on relevant attitudes,…

  20. Permissive norms and young adults' alcohol and marijuana use: the role of online communities.

    PubMed

    Stoddard, Sarah A; Bauermeister, Jose A; Gordon-Messer, Deborah; Johns, Michelle; Zimmerman, Marc A

    2012-11-01

    Young adults are increasingly interacting with their peer groups online through social networking sites. These online interactions may reinforce or escalate alcohol and other drug (AOD) use as a result of more frequent and continuous exposure to AOD promotive norms; however, the influence of young adults' virtual networks on AOD use remains untested. The purpose of this study was to examine the association between the presence of AOD use content in online social networking, perceived norms (online norms regarding AOD use and anticipated regret with AOD use postings), and alcohol and marijuana use in a sample of 18- to 24-year-olds. Using an adapted web version of respondent-driven sampling (webRDS), we recruited a sample of 18- to 24-year-olds (N = 3,448) in the United States. Using multivariate regression, we explored the relationship between past-30-day alcohol and marijuana use, online norms regarding AOD use, peer substance use, and online and offline peer support. Alcohol use was associated with more alcohol content online. Anticipated regret and online peer support were associated with less alcohol use. Anticipated regret was negatively associated with marijuana use. Peer AOD use was positively associated with both alcohol and marijuana use. Peers play an important role in young adult alcohol and marijuana use, whether online or in person. Our findings highlight the importance of promoting online network-based AOD prevention programs for young adults in the United States.

  1. Steady-state generation of hydrogen peroxide: kinetics and stability of alcohol oxidase immobilized on nanoporous alumina.

    PubMed

    Kjellander, Marcus; Götz, Kathrin; Liljeruhm, Josefine; Boman, Mats; Johansson, Gunnar

    2013-04-01

    Alcohol oxidase from Pichia pastoris was immobilized on nanoporous aluminium oxide membranes by silanization and activation by carbonyldiimidazole to create a flow-through enzyme reactor. Kinetic analysis of the hydrogen peroxide generation was carried out for a number of alcohols using a subsequent reaction with horseradish peroxidase and ABTS. The activity data for the immobilized enzyme showed a general similarity with literature data in solution, and the reactor could generate 80 mmol H2O2/h per litre reactor volume. Horseradish peroxidase was immobilized by the same technique to construct bienzymatic modular reactors. These were used in both single pass mode and circulating mode. Pulsed injections of methanol resulted in a linear relation between response and concentration, allowing quantitative concentration measurement. The immobilized alcohol oxidase retained 58 % of initial activity after 3 weeks of storage and repeated use.

  2. Performance of optical biosensor using alcohol oxidase enzyme for formaldehyde detection

    NASA Astrophysics Data System (ADS)

    Sari, A. P.; Rachim, A.; Nurlely, Fauzia, V.

    2017-07-01

    The recent issue in the world is the long exposure of formaldehyde which is can increase the risk of human health, therefore, that is very important to develop a device and method that can be optimized to detect the formaldehyde elements accurately, have a long lifetime and can be fabricated and produced in large quantities. A new and simple prepared optical biosensor for detection of formaldehyde in aqueous solutions using alcohol oxidase (AOX) enzyme was successfully fabricated. The poly-n-butyl acrylic-co-N-acryloxysuccinimide (nBA-NAS) membranes containing chromoionophore ETH5294 were used for immobilization of alcohol oxidase enzyme (AOX). Biosensor response was based on the colour change of chromoionophore as a result of enzymatic oxidation of formaldehyde and correlated with the detection concentration of formaldehyde. The performance of biosensor parameters were measured through the optical absorption value using UV-Vis spectrophotometer including the repeatability, reproducibility, selectivity and lifetime. The results showed that the prepared biosensor has good repeatability (RSD = 1.9 %) and good reproducibility (RSD = 2.1 %). The biosensor was selective formaldehyde with no disturbance by methanol, ethanol, and acetaldehyde, and also stable before 49 days and decrease by 41.77 % after 49 days.

  3. Fungal aryl-alcohol oxidase: a peroxide-producing flavoenzyme involved in lignin degradation.

    PubMed

    Hernández-Ortega, Aitor; Ferreira, Patricia; Martínez, Angel T

    2012-02-01

    Aryl-alcohol oxidase (AAO) is an extracellular flavoprotein providing the H(2)O(2) required by ligninolytic peroxidases for fungal degradation of lignin, the key step for carbon recycling in land ecosystems. O(2) activation by Pleurotus eryngii AAO takes place during the redox-cycling of p-methoxylated benzylic metabolites secreted by the fungus. Only Pleurotus AAO sequences were available for years, but the number strongly increased recently due to sequencing of different basidiomycete genomes, and a comparison of 112 GMC (glucose-methanol-choline oxidase) superfamily sequences including 40 AAOs is presented. As shown by kinetic isotope effects, alcohol oxidation by AAO is produced by hydride transfer to the flavin, and hydroxyl proton transfer to a base. Moreover, site-directed mutagenesis studies showed that His502 activates the alcohol substrate by proton abstraction, and this result was extended to other GMC oxidoreductases where the nature of the base was under discussion. However, in contrast with that proposed for GMC oxidoreductases, the two transfers are not stepwise but concerted. Alcohol docking at the buried AAO active site resulted in only one catalytically relevant position for concerted transfer, with the pro-R α-hydrogen at distance for hydride abstraction. The expected hydride-transfer stereoselectivity was demonstrated, for the first time in a GMC oxidoreductase, by using the (R) and (S) enantiomers of α-deuterated p-methoxybenzyl alcohol. Other largely unexplained aspects of AAO catalysis (such as the unexpected specificity on substituted aldehydes) can also be explained in the light of the recent results. Finally, the biotechnological interest of AAO in flavor production is extended by its potential in production of chiral compounds taking advantage from the above-described stereoselectivity.

  4. Analysis of monoamine oxidase A (MAOA) promoter polymorphism in Finnish male alcoholics.

    PubMed

    Saito, Takuya; Lachman, Herbert M; Diaz, Libna; Hallikainen, Tero; Kauhanen, Jussi; Salonen, Jukka T; Ryynänen, Olli-Pekka; Karvonen, Matti K; Syvälahti, Erkka; Pohjalainen, Tiina; Hietala, Jarmo; Tiihonen, Jari

    2002-03-15

    Alterations in monoamine oxidase A (MAOA) expression and enzyme activity may be associated with alcoholism and impulsive behavior. Therefore, functional polymorphisms in the MAOA gene would be good candidates to consider in the interindividual differences that exist in the susceptibility to alcoholism. One variant that has been considered as a candidate in alcoholism is a repeat polymorphism in the MAOA gene promoter. We analyzed a cohort of Finnish males with either type 1 or type 2 alcoholism, as well as controls, for differences in the distribution of MAOA promoter alleles. Based on other studies, we postulated that type 2 alcoholism, which is associated with antisocial behavior, but not type 1 alcoholism, would be correlated with the inheritance of the low promoter activity allele. However, we failed to find a difference in allele distribution in type 1 and type 2 alcoholics. In addition, there was no difference in the allele distribution when each group of alcoholics was compared with controls. However, when both groups of alcoholics were pooled and compared with controls, the difference in allele distribution reached a trend towards significance. Our results suggest a minimal association between the MAOA low activity promoter alleles and alcoholism, regardless of the presence or absence of antisocial behavior. Interestingly, approximately 3% of type 2 alcoholics were found to be heterozygous for the MAOA promoter polymorphism. Since MAOA is X-linked, the heterozygotes are probable cases of Klinefelter's syndrome (47,XXY) suggesting that X-chromosome aneuploidy may increase the risk for developing type 2 alcoholism.

  5. Structure–function characterization reveals new catalytic diversity in the galactose oxidase and glyoxal oxidase family

    PubMed Central

    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

    2015-01-01

    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. PMID:26680532

  6. Automated Orbit Determination System (AODS) requirements definition and analysis

    NASA Technical Reports Server (NTRS)

    Waligora, S. R.; Goorevich, C. E.; Teles, J.; Pajerski, R. S.

    1980-01-01

    The requirements definition for the prototype version of the automated orbit determination system (AODS) is presented including the AODS requirements at all levels, the functional model as determined through the structured analysis performed during requirements definition, and the results of the requirements analysis. Also specified are the implementation strategy for AODS and the AODS-required external support software system (ADEPT), input and output message formats, and procedures for modifying the requirements.

  7. Cross-Lagged Associations Between Substance Use-Related Media Exposure and Alcohol Use During Middle School

    PubMed Central

    Tucker, Joan S.; Miles, Jeremy N. V.; D’Amico, Elizabeth J.

    2013-01-01

    Purpose This study examines the reciprocal longitudinal associations between alcohol or other drug (AOD)-related media exposure and alcohol use among middle school students, and explores whether these associations differ by ethnicity or gender. Methods The analytic sample is 7th grade students who were recruited from 16 California middle schools and surveyed in the spring semester of two academic years. Students reported on their background characteristics, exposure to seven types of AOD-related media content (internet videos, social networking sites, movies, television, magazine advertisements, songs, and video games) in the past 3 months, and alcohol use in the past 30 days. Structural equation modeling was used to examine cross-lagged associations between media exposure and alcohol use. Results Greater AOD-related media exposure in 7th grade was significantly associated with a higher probability of alcohol use in 8th grade (p=.02), and alcohol use in 7th grade was marginally associated with greater AOD-related media exposure in 8th grade (p=.07). These cross-lagged associations did not statistically differ by ethnicity (Hispanic vs. non-Hispanic white) or gender. Further, there was no evidence that certain types of media exposure were more strongly associated with alcohol use than others. Conclusions Results from this study suggest that AOD-related media effects and media selectively form a reciprocal, mutually influencing process that may escalate adolescent alcohol use over time. Addressing adolescents’ exposure to AOD-related media content and its effects on behavior, such as through media literacy education, may hold promise for improving the efficacy of alcohol prevention efforts for middle school students. PMID:23770074

  8. Cross-lagged associations between substance use-related media exposure and alcohol use during middle school.

    PubMed

    Tucker, Joan S; Miles, Jeremy N V; D'Amico, Elizabeth J

    2013-10-01

    This study examines the reciprocal longitudinal associations between alcohol or other drug (AOD)-related media exposure and alcohol use among middle school students, and explores whether these associations differ by ethnicity or gender. The analytic sample is 7th grade students who were recruited from 16 California middle schools and surveyed in the spring semester of two academic years. Students reported on their background characteristics, exposure to seven types of AOD-related media content (Internet videos, social networking sites, movies, television, magazine advertisements, songs, and video games) in the past 3 months, and alcohol use in the past 30 days. Structural equation modeling was used to examine cross-lagged associations between media exposure and alcohol use. Greater AOD-related media exposure in 7th grade was significantly associated with a higher probability of alcohol use in 8th grade (p = .02), and alcohol use in 7th grade was marginally associated with greater AOD-related media exposure in 8th grade (p = .07). These cross-lagged associations did not statistically differ by ethnicity (Hispanic vs. non-Hispanic white) or gender. Further, there was no evidence that certain types of media exposure were more strongly associated with alcohol use than others. Results from this study suggest that AOD-related media effects and media selectively form a reciprocal, mutually influencing process that may escalate adolescent alcohol use over time. Addressing adolescents' exposure to AOD-related media content and its effects on behavior, such as through media literacy education, may hold promise for improving the efficacy of alcohol prevention efforts for middle school students. Copyright © 2013 Society for Adolescent Health and Medicine. Published by Elsevier Inc. All rights reserved.

  9. Insomnia, platelet serotonin and platelet monoamine oxidase in chronic alcoholism.

    PubMed

    Nenadic Sviglin, Korona; Nedic, Gordana; Nikolac, Matea; Mustapic, Maja; Muck-Seler, Dorotea; Borovecki, Fran; Pivac, Nela

    2011-08-18

    Insomnia is a common sleep disorder frequently occurring in chronic alcoholic patients. Neurobiological basis of insomnia, as well as of alcoholism, is associated with disrupted functions of the main neurotransmitter systems, including the serotonin (5-hydroxytryptamine, 5-HT) system. Blood platelets are considered a limited peripheral model for the central 5-HT neurons, since both platelets and central 5-HT synaptosomes have similar dynamics of 5-HT. Platelet 5-HT concentration and platelet monoamine oxidase type B (MAO-B) are assumed to represent biomarkers for particular symptoms and behaviors in psychiatric disorders. The hypothesis of this study was that platelet 5-HT concentration and platelet MAO-B activity will be altered in chronic alcoholic patients with insomnia compared to comparable values in patients without insomnia. The study included 498 subjects: 395 male and 103 female medication-free patients with alcohol dependence and 502 healthy control subjects: 325 men and 177 women. The effects of early, middle and late insomnia (evaluated using the Hamilton Depression Rating Scale), as well as sex, age and smoking on platelet 5-HT concentration and platelet MAO-B activity were evaluated using one-way ANOVA and multiple regression analysis by the stepwise method. Platelet 5-HT concentration, but not platelet MAO-B activity, was significantly reduced in alcoholic patients with insomnia compared to patients without insomnia. Multiple regression analysis revealed that platelet 5-HT concentration was affected by middle insomnia, smoking and sex, while platelet MAO activity was affected only by sex and age. The present and previous data suggest that platelet 5-HT concentration might be used, after controlling for sex and smoking, as a biomarker for insomnia in alcoholism, PTSD and in rotating shift workers. Copyright © 2011 Elsevier Ireland Ltd. All rights reserved.

  10. A novel amperometric alcohol biosensor developed in a 3rd generation bioelectrode platform using peroxidase coupled ferrocene activated alcohol oxidase as biorecognition system.

    PubMed

    Chinnadayyala, Somasekhar R; Kakoti, Ankana; Santhosh, Mallesh; Goswami, Pranab

    2014-05-15

    Alcohol oxidase (AOx) with a two-fold increase in efficiency (Kcat/Km) was achieved by physical entrapment of the activator ferrocene in the protein matrix through a simple microwave based partial unfolding technique and was used to develop a 3rd generation biosensor for improved detection of alcohol in liquid samples. The ferrocene molecules were stably entrapped in the AOx protein matrix in a molar ratio of ~3:1 through electrostatic interaction with the Trp residues involved in the functional activity of the enzyme as demonstrated by advanced analytical techniques. The sensor was fabricated by immobilizing ferrocene entrapped alcohol oxidase (FcAOx) and sol-gel chitosan film coated horseradish peroxidase (HRP) on a multi-walled carbon nanotube (MWCNT) modified glassy carbon electrode through layer-by-layer technique. The bioelectrode reactions involved the formation of H2O2 by FcAOx biocatalysis of substrate alcohol followed by HRP-catalyzed reduction of the liberated H2O2 through MWCNT supported direct electron transfer mechanism. The amperometric biosensor exhibited a linear response to alcohol in the range of 5.0 × 10(-6) to 30 × 10(-4)mol L(-1) with a detection limit of 2.3 × 10(-6) mol L(-1), and a sensitivity of 150 µA mM(-1) cm(-2). The biosensor response was steady for 28 successive measurements completed in a period of 5h and retained ~90% of the original response even after four weeks when stored at 4 °C. The biosensor was successfully applied for the determination of alcohol in commercial samples and its performance was validated by comparing with the data obtained by GC analyses of the samples. © 2013 Published by Elsevier B.V.

  11. Portal Surveys of Time-Out Drinking Locations: A Tool for Studying Binge Drinking and AOD Use

    ERIC Educational Resources Information Center

    Voas, Robert B.; Furr-Holden, Debra; Lauer, Elizabeth; Bright, Kristin; Johnson, Mark B.; Miller, Brenda

    2006-01-01

    Portal surveys, defined as assessments occurring proximal to the entry point to a high-risk locale and immediately on exit, can be used in different settings to measure characteristics and behavior of attendees at an event of interest. This methodology has been developed to assess alcohol and other drug (AOD) use at specific events and has…

  12. Combination of polymorphic variants in serotonin transporter and monoamine oxidase-A genes may influence the risk for early-onset alcoholism.

    PubMed

    Bordukalo-Niksic, Tatjana; Stefulj, Jasminka; Matosic, Ana; Mokrovic, Gordana; Cicin-Sain, Lipa

    2012-12-30

    The combinatory effect of polymorphisms in serotonin transporter and monoamine oxidase-A genes on the aetiopathogenesis of alcoholism was investigated in a sample of 714 individuals. Increased frequency of subjects having three 'suspected' genotypes (5-HTTLPR-LL, STin2-1010 and MAO-A 3-repeat allele) was found among type-2 alcoholic patients (P=0.0189). Results highlight serotonergic/genetic contribution to early-onset alcoholism. Copyright © 2012 Elsevier Ireland Ltd. All rights reserved.

  13. Health Professionals' Attitudes towards AOD-Related Work: Moving the Traditional Focus from Education and Training to Organizational Culture

    ERIC Educational Resources Information Center

    Skinner, Natalie; Roche, Ann M.; Freeman, Toby; Mckinnon, Anna

    2009-01-01

    Aim: This article presents a critical review of research on health professionals' attitudes towards alcohol and other drug (AOD)-related work relevant to both researchers and practitioners. It moves beyond education and training programs to examine the relevance of organizational culture in influencing attitudes. Method: A review of research…

  14. Characterization of an aryl-alcohol oxidase from the plant saprophytic basidiomycete Coprinopsis cinerea with broad substrate specificity against aromatic alcohols.

    PubMed

    Tamaru, Yoshiaki; Umezawa, Kiwamu; Yoshida, Makoto

    2018-07-01

    The aim of the study was to obtain information about the enzymatic properties of aryl-alcohol oxidase from the plant saprophytic basidiomycete Coprinopsis cinerea (rCcAAO), which is classified into the auxiliary activities family 3 subfamily 2 (AA3_2). The gene encoding AAO from the plant saprophytic basidiomycete Coprinopsis cinerea (CcAAO) was cloned, and the recombinant CcAAO (rCcAAO) was heterologously expressed in the methylotrophic yeast Pichia pastoris. The purified rCcAAO showed significant activity not only against trans,trans-2,4-hexadien-1-ol but also against a broad range of aromatic alcohols including aromatic compounds that were reported to be poor substrates for known AAOs. Moreover, site-directed mutagenesis analysis demonstrated that mutants with substitutions from leucine to phenylalanine and tryptophan at position 416 exhibited decreases of activity for aromatic alcohols but still maintained the activity for trans,trans-2,4-hexadien-1-ol. Leucine 416 in CcAAO contributes to the broad substrate specificity against various aromatic alcohols, which is useful for the production of hydrogen peroxide using this enzyme.

  15. Bayesian aerosol retrieval algorithm for MODIS AOD retrieval over land

    NASA Astrophysics Data System (ADS)

    Lipponen, Antti; Mielonen, Tero; Pitkänen, Mikko R. A.; Levy, Robert C.; Sawyer, Virginia R.; Romakkaniemi, Sami; Kolehmainen, Ville; Arola, Antti

    2018-03-01

    We have developed a Bayesian aerosol retrieval (BAR) algorithm for the retrieval of aerosol optical depth (AOD) over land from the Moderate Resolution Imaging Spectroradiometer (MODIS). In the BAR algorithm, we simultaneously retrieve all dark land pixels in a granule, utilize spatial correlation models for the unknown aerosol parameters, use a statistical prior model for the surface reflectance, and take into account the uncertainties due to fixed aerosol models. The retrieved parameters are total AOD at 0.55 µm, fine-mode fraction (FMF), and surface reflectances at four different wavelengths (0.47, 0.55, 0.64, and 2.1 µm). The accuracy of the new algorithm is evaluated by comparing the AOD retrievals to Aerosol Robotic Network (AERONET) AOD. The results show that the BAR significantly improves the accuracy of AOD retrievals over the operational Dark Target (DT) algorithm. A reduction of about 29 % in the AOD root mean square error and decrease of about 80 % in the median bias of AOD were found globally when the BAR was used instead of the DT algorithm. Furthermore, the fraction of AOD retrievals inside the ±(0.05+15 %) expected error envelope increased from 55 to 76 %. In addition to retrieving the values of AOD, FMF, and surface reflectance, the BAR also gives pixel-level posterior uncertainty estimates for the retrieved parameters. The BAR algorithm always results in physical, non-negative AOD values, and the average computation time for a single granule was less than a minute on a modern personal computer.

  16. Simultaneous and accurate real-time monitoring of glucose and ethanol in alcoholic drinks, must, and biomass by a dual-amperometric biosensor.

    PubMed

    Mentana, Annalisa; Palermo, Carmen; Nardiello, Donatella; Quinto, Maurizio; Centonze, Diego

    2013-01-09

    In this work the optimization and application of a dual-amperometric biosensor for simultaneous monitoring of glucose and ethanol content, as quality markers in drinks and alcoholic fermentation media, are described. The biosensor is based on glucose oxidase (GOD) and alcohol oxidase (AOD) immobilized by co-cross-linking with bovine serum albumin (BSA) and glutaraldehyde (GLU) both onto a dual gold electrode, modified with a permselective overoxidized polypyrrole film (PPYox). Response, rejection of interferents, and stability of the dual biosensor were optimized in terms of PPYox thickness, BSA, and enzyme loading. The biosensor was integrated in a flow injection system coupled with an at-line microdialysis fiber as a sampling tool. Flow rates inside and outside the fiber were optimized in terms of linear responses (0.01-1 and 0.01-1.5 M) and sensitivities (27.6 ± 0.4 and 31.0 ± 0.6 μA·M(-1)·cm(-2)) for glucose and ethanol. Excellent anti-interference characteristics, the total absence of "cross-talk", and good response stability under operational conditions allowed application of the dual biosensor in accurate real-time monitoring (at least 15 samples/h) of alcoholic drinks, white grape must, and woody biomass.

  17. In cellulo serial crystallography of alcohol oxidase crystals inside yeast cells

    PubMed Central

    Jakobi, Arjen J.; Passon, Daniel M.; Knoops, Kèvin; Stellato, Francesco; Liang, Mengning; White, Thomas A.; Seine, Thomas; Messerschmidt, Marc; Chapman, Henry N.; Wilmanns, Matthias

    2016-01-01

    The possibility of using femtosecond pulses from an X-ray free-electron laser to collect diffraction data from protein crystals formed in their native cellular organelle has been explored. X-ray diffraction of submicrometre-sized alcohol oxidase crystals formed in peroxisomes within cells of genetically modified variants of the methylotrophic yeast Hansenula polymorpha is reported and characterized. The observations are supported by synchrotron radiation-based powder diffraction data and electron microscopy. Based on these findings, the concept of in cellulo serial crystallography on protein targets imported into yeast peroxisomes without the need for protein purification as a requirement for subsequent crystallization is outlined. PMID:27006771

  18. In cellulo serial crystallography of alcohol oxidase crystals inside yeast cells

    DOE PAGES

    Jakobi, Arjen J.; Passon, Daniel M.; Knoops, Kevin; ...

    2016-03-01

    The possibility of using femtosecond pulses from an X-ray free-electron laser to collect diffraction data from protein crystals formed in their native cellular organelle has been explored. X-ray diffraction of submicrometre-sized alcohol oxidase crystals formed in peroxisomes within cells of genetically modified variants of the methylotrophic yeast Hansenula polymorpha is reported and characterized. Furthermore, the observations are supported by synchrotron radiation-based powder diffraction data and electron microscopy. Based on these findings, the concept of in cellulo serial crystallography on protein targets imported into yeast peroxisomes without the need for protein purification as a requirement for subsequent crystallization is outlined.

  19. In cellulo serial crystallography of alcohol oxidase crystals inside yeast cells

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

    Jakobi, Arjen J.; Passon, Daniel M.; Knoops, Kevin

    The possibility of using femtosecond pulses from an X-ray free-electron laser to collect diffraction data from protein crystals formed in their native cellular organelle has been explored. X-ray diffraction of submicrometre-sized alcohol oxidase crystals formed in peroxisomes within cells of genetically modified variants of the methylotrophic yeast Hansenula polymorpha is reported and characterized. Furthermore, the observations are supported by synchrotron radiation-based powder diffraction data and electron microscopy. Based on these findings, the concept of in cellulo serial crystallography on protein targets imported into yeast peroxisomes without the need for protein purification as a requirement for subsequent crystallization is outlined.

  20. Monthly AOD maps combining strengths of remote sensing products

    NASA Astrophysics Data System (ADS)

    Kinne, Stefan

    2010-05-01

    The mid-visible aerosol optical depth (AOD) is the most prominent property to quantify aerosol amount the atmospheric column. Almost all aerosol retrievals of satellite sensors provide estimates for this property, however, often with limited success. As sensors differ in capabilities individual retrievals have local and regional strengths and weaknesses. Focusing on individual retrieval strengths a satellite based AOD composite has been constructed. Hereby, every retrieval performance has been assessed in statistical comparisons to ground-based sun-photometry, which provide highly accurate references though only at few globally distributed monitoring sites. Based on these comparisons, which consider bias as well as spatial patterns and seasonality, the regionally best performing satellite AOD products are combined. The resulting remote sensing AOD composite provide a general reference for the spatial and temporal AOD distribution on an (almost) global basis - solely tied to sensor data.

  1. Evaluating the Impact of Hospital Based Drug and Alcohol Consultation Liaison Services.

    PubMed

    Reeve, Rebecca; Arora, Sheena; Butler, Kerryn; Viney, Rosalie; Burns, Lucinda; Goodall, Stephen; van Gool, Kees

    2016-09-01

    Consultation liaison (CL) services provide direct access to specialist services for support, treatment advice and assistance with the management of a given condition. Alcohol and other drugs (AOD) CL services aim to improve identification and treatment of patients with AOD morbidity. Our objective was to evaluate the costs and consequences of AOD CL services in hospitals in New South Wales, Australia. Patients were surveyed at eight hospitals and problematic AOD use was identified using the Alcohol, Smoking and Substance Involvement Screening Test (n=1615). For consenting participants, medical record data were obtained from 18 months pre- to 12 months post-survey. We used interrupted time series analyses to compare utilization and costs for patients with and without AOD problems and changes over time between those who received AOD CL and similar patients. Approximately 35% of patients surveyed had AOD problems (excluding tobacco) with 7% requiring intensive treatment. Only 24% of patients requiring intensive treatment were treated by AOD CL. Those treated had relative improvements over time in the cost of presentations to emergency departments, emergency admission performance and increased uptake of appropriate pharmaceuticals. The estimated net benefit of AOD CL services was at least AUD$100,000 savings per hospital per year. Expanding AOD CL services to address current unmet need may lead to even greater cost savings for hospitals. Copyright © 2016 Elsevier Inc. All rights reserved.

  2. Structure of Alcohol Oxidase from Pichia pastoris by Cryo-Electron Microscopy

    PubMed Central

    Vonck, Janet; Parcej, David N.; Mills, Deryck J.

    2016-01-01

    The first step in methanol metabolism in methylotrophic yeasts, the oxidation of methanol and higher alcohols with molecular oxygen to formaldehyde and hydrogen peroxide, is catalysed by alcohol oxidase (AOX), a 600-kDa homo-octamer containing eight FAD cofactors. When these yeasts are grown with methanol as the carbon source, AOX forms large crystalline arrays in peroxisomes. We determined the structure of AOX by cryo-electron microscopy at a resolution of 3.4 Å. All residues of the 662-amino acid polypeptide as well as the FAD are well resolved. AOX shows high structural homology to other members of the GMC family of oxidoreductases, which share a conserved FAD binding domain, but have different substrate specificities. The preference of AOX for small alcohols is explained by the presence of conserved bulky aromatic residues near the active site. Compared to the other GMC enzymes, AOX contains a large number of amino acid inserts, the longest being 75 residues. These segments are found at the periphery of the monomer and make extensive inter-subunit contacts which are responsible for the very stable octamer. A short surface helix forms contacts between two octamers, explaining the tendency of AOX to form crystals in the peroxisomes. PMID:27458710

  3. An "In Situ" Calibration Correction Procedure (KCICLO) Based on AOD Diurnal Cycle: Application to AERONET-El Arenosillo (Spain) AOD Data Series

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

    Cachorro, V. E.; Toledano, C.; Berjon, A.

    Aerosol optical depth (AOD) very often shows a distinct diurnal cycle pattern, which seems to be an artifact resulting from an incorrect calibration (or an equivalent effect, such as filter degradation). The shape of this fictitious AOD diurnal cycle varies as the inverse of the solar air mass (m) and the magnitude of the effect is greatest at midday. The observation of this effect is not easy at many field stations, and only those stations with good weather conditions permit an easier detection and the possibility of its correction. By taking advantage of this dependence on the air mass, wemore » propose an improved “in situ” correction-calibration procedure to AOD measured data series. The method is named KCICLO because the determination of a constant K and the behavior of AOD as a cycle (ciclo, in Spanish). We estimate it has an accuracy of 0.2–0.5% for the calibration ratio constant K, or 0.002–0.005 in AOD at field stations. Although the KCICLO is an “in situ” calibration method, we recommend it to be used as an AOD correction method for field stations. At high-altitude sites, it may be used independently of the classical Langley method (CLM). However, we also recommend it to be used as a complement to CLM, improving it considerably. The application of this calibration correction method to the nearly 5 year AOD data series at El Arenosillo (Huelva, southwestern Spain) station belonging to Aerosol Robotic Network (AERONET)-PHOTONS shows that 8 (50%) of 16 filters of the four analyzed Sun photometers were outside of the 0.02 uncertainty of AERONET specification. The largest departures reached values of 0.06. The results show the efficiency of the method and a significant improvement over other “in situ” methods, with no other information required beyond the same AOD data.« less

  4. Molecular Characterization and Expression of a Novel Alcohol Oxidase from Aspergillus terreus MTCC6324

    PubMed Central

    Chakraborty, Mitun; Goel, Manish; Chinnadayyala, Somasekhar R.; Dahiya, Ujjwal Ranjan; Ghosh, Siddhartha Sankar; Goswami, Pranab

    2014-01-01

    The alcohol oxidase (AOx) cDNA from Aspergillus terreus MTCC6324 with an open reading frame (ORF) of 2001 bp was constructed from n-hexadecane induced cells and expressed in Escherichia coli with a yield of ∼4.2 mg protein g−1 wet cell. The deduced amino acid sequences of recombinant rAOx showed maximum structural homology with the chain B of aryl AOx from Pleurotus eryngii. A functionally active AOx was achieved by incubating the apo-AOx with flavin adenine dinucleotide (FAD) for ∼80 h at 16°C and pH 9.0. The isoelectric point and mass of the apo-AOx were found to be 6.5±0.1 and ∼74 kDa, respectively. Circular dichroism data of the rAOx confirmed its ordered structure. Docking studies with an ab-initio protein model demonstrated the presence of a conserved FAD binding domain with an active substrate binding site. The rAOx was specific for aryl alcohols and the order of its substrate preference was 4-methoxybenzyl alcohol >3-methoxybenzyl alcohol>3, 4-dimethoxybenzyl alcohol > benzyl alcohol. A significantly high aggregation to ∼1000 nm (diameter) and catalytic efficiency (kcat/Km) of 7829.5 min−1 mM−1 for 4-methoxybenzyl alcohol was also demonstrated for rAOx. The results infer the novelty of the AOx and its potential biocatalytic application. PMID:24752075

  5. Molecular characterization and expression of a novel alcohol oxidase from Aspergillus terreus MTCC6324.

    PubMed

    Chakraborty, Mitun; Goel, Manish; Chinnadayyala, Somasekhar R; Dahiya, Ujjwal Ranjan; Ghosh, Siddhartha Sankar; Goswami, Pranab

    2014-01-01

    The alcohol oxidase (AOx) cDNA from Aspergillus terreus MTCC6324 with an open reading frame (ORF) of 2001 bp was constructed from n-hexadecane induced cells and expressed in Escherichia coli with a yield of ∼4.2 mg protein g-1 wet cell. The deduced amino acid sequences of recombinant rAOx showed maximum structural homology with the chain B of aryl AOx from Pleurotus eryngii. A functionally active AOx was achieved by incubating the apo-AOx with flavin adenine dinucleotide (FAD) for ∼80 h at 16°C and pH 9.0. The isoelectric point and mass of the apo-AOx were found to be 6.5±0.1 and ∼74 kDa, respectively. Circular dichroism data of the rAOx confirmed its ordered structure. Docking studies with an ab-initio protein model demonstrated the presence of a conserved FAD binding domain with an active substrate binding site. The rAOx was specific for aryl alcohols and the order of its substrate preference was 4-methoxybenzyl alcohol >3-methoxybenzyl alcohol>3, 4-dimethoxybenzyl alcohol > benzyl alcohol. A significantly high aggregation to ∼1000 nm (diameter) and catalytic efficiency (kcat/Km) of 7829.5 min-1 mM-1 for 4-methoxybenzyl alcohol was also demonstrated for rAOx. The results infer the novelty of the AOx and its potential biocatalytic application.

  6. The Impact of Alcohol and Other Drugs on the Child Welfare System.

    ERIC Educational Resources Information Center

    Curtis, Patrick A.; McCullough, Charlotte

    1993-01-01

    Surveyed Child Welfare League of America member agencies about the impact of alcohol and other drugs (AOD) on child welfare services delivery. Agencies reported that AOD adversely affected the system by compounding problems, such as personnel shortages and shortages in available foster homes, which existed before AOD was identified as a major…

  7. AOD trends during 2001-2010 from observations and model simulations

    NASA Astrophysics Data System (ADS)

    Pozzer, A.; de Meij, A.; Yoon, J.; Tost, H.; Georgoulias, A. K.; Astitha, M.

    2015-05-01

    The aerosol optical depth (AOD) trend between 2001 and 2010 is estimated globally and regionally from observations and results from simulations with the EMAC (ECHAM5/MESSy Atmospheric Chemistry) model. Although interannual variability is applied only to anthropogenic and biomass-burning emissions, the model is able to quantitatively reproduce the AOD trends as observed by the MODIS (Moderate Resolution Imaging Spectroradiometer) satellite sensor, while some discrepancies are found when compared to MISR (Multi-angle Imaging SpectroRadiometer) and SeaWIFS (Sea-viewing Wide Field-of-view Sensor) observations. Thanks to an additional simulation without any change in emissions, it is shown that decreasing AOD trends over the US and Europe are due to the decrease in the emissions, while over the Sahara Desert and the Middle East region, the meteorological changes play a major role. Over Southeast Asia, both meteorology and emissions changes are equally important in defining AOD trends. Additionally, decomposing the regional AOD trends into individual aerosol components reveals that the soluble components are the most dominant contributors to the total AOD, as their influence on the total AOD is enhanced by the aerosol water content.

  8. Hydride transfer made easy in the oxidation of alcohols catalyzed by choline oxidase

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

    Gadda, G.; Orville, A.; Pennati, A.

    2008-06-08

    Choline oxidase (E.C. 1.1.3.17) catalyzes the two-step, four-electron oxidation of choline to glycine betaine with betaine aldehyde as enzyme-associated intermediate and molecular oxygen as final electron acceptor (Scheme 1). The gem-diol, hydrated species of the aldehyde intermediate of the reaction acts as substrate for aldehyde oxidation, suggesting that the enzyme may use similar strategies for the oxidation of the alcohol substrate and aldehyde intermediate. The determination of the chemical mechanism for alcohol oxidation has emerged from biochemical, mechanistic, mutagenetic, and structural studies. As illustrated in the mechanism of Scheme 2, the alcohol substrate is initially activated in the active sitemore » of the enzyme by removal of the hydroxyl proton. The resulting alkoxide intermediate is then stabilized in the enzyme-substrate complex via electrostatic interactions with active site amino acid residues. Alcohol oxidation then occurs quantum mechanically via the transfer of the hydride ion from the activated substrate to the N(5) flavin locus. An essential requisite for this mechanism of alcohol oxidation is the high degree of preorganization of the activated enzyme-substrate complex, which is achieved through an internal equilibrium of the Michaelis complex occurring prior to, and independently from, the subsequent hydride transfer reaction. The experimental evidence that support the mechanism for alcohol oxidation shown in Scheme 2 is briefly summarized in the Results and Discussion section.« less

  9. The GEOS-5 Neural Network Retrieval for AOD

    NASA Astrophysics Data System (ADS)

    Castellanos, P.; da Silva, A. M., Jr.

    2017-12-01

    One of the difficulties in data assimilation is the need for multi-sensor data merging that can account for temporal and spatial biases between satellite sensors. In the Goddard Earth Observing System Model Version 5 (GEOS-5) aerosol data assimilation system, a neural network retrieval (NNR) is used as a mapping between satellite observed top of the atmosphere (TOA) reflectance and AOD, which is the target variable that is assimilated in the model. By training observations of TOA reflectance from multiple sensors to map to a common AOD dataset (in this case AOD observed by the ground based Aerosol Robotic Network, AERONET), we are able to create a global, homogenous, satellite data record of AOD from MODIS observations on board the Terra and Aqua satellites. In this talk, I will present the implementation of and recent updates to the GEOS-5 NNR for MODIS collection 6 data.

  10. Validation and Comparison of AATRS AOD L2 Products over China

    NASA Astrophysics Data System (ADS)

    Che, Yahui; Xue, Yong; Guang, Jie; Guo, Jianping; Li, Ying

    2016-04-01

    The Advanced Along-Track Scanning Radiometer (AATSR) aboard on ENVISAT has been used to observe the Earth for more than 10 years since than 2002. One of main applications of AATSR instrument is to observe atmospheric aerosol, especially in retrieval of aerosol optical depth (AOD), taking advantage of its dual-view that helps to separate the contribution of aerosol from top of atmosphere reflectance (A. A. Kokhanovsky and de Leeuw, 2009). The project of Aerosol_CCI, as part of European Space Agency's Climate Change Initiative (CCI), has released new AATSR aerosol AOD products by the of 2015, including the SU v4.21 product from Swansea algorithm, ADV v2.3 product from the ATSR-2/AATSR dual view aerosol retrieval algorithm (ADV) and ORAC v03.04 product from the Oxford-RAL Retrieval of Aerosol and Cloud algorithm. The previous versions of these three AOD level 2 (L2) products in 2008 have been validated over mainland China (Che and Xue, 2015). In this paper, we validated these AATSR AOD products with latest versions in mainland China in 2007, 2008 and 2010 by the means of comparison with the AErosol RObotic NETwork (AERONET) and the China Aerosol Remote Sensing Network (CARSNET). The combination of AERONET and CARSNET helps to make up for the disadvantages of small number and uneven distribution of AEROENT cites. The validation results show different performance of these AOD products over China. The performances of SU and ADV products seem to be the same with close correlation coefficient (CC) about 08~0.9 and root mean square (RMS) within 0.15 in all three years, and sensitive to high AOD values (AOD >1): more AODs and more underestimated. However, these two products do exist difference, which is that the SU algorithm retrieves more high AODs, leading to more space-time validation matches with ground-based data. The ORAC algorithm is different from the others, it can be not only used to retrieve low AODs but also high AODs over different landcover types. Even though

  11. "What constitutes a 'problem'?" Producing 'alcohol problems' through online counselling encounters.

    PubMed

    Savic, Michael; Ferguson, Nyssa; Manning, Victoria; Bathish, Ramez; Lubman, Dan I

    2017-08-01

    Typically, health policy, practice and research views alcohol and other drug (AOD) 'problems' as objective things waiting to be detected, diagnosed and treated. However, this approach to policy development and treatment downplays the role of clinical practices, tools, discourses, and systems in shaping how AOD use is constituted as a 'problem'. For instance, people might present to AOD treatment with multiple psycho-social concerns, but usually only a singular AOD-associated 'problem' is considered serviceable. As the assumed nature of 'the serviceable problem' influences what treatment responses people receive, and how they may come to be enacted as 'addicted' or 'normal' subjects, it is important to subject clinical practices of problem formulation to critical analysis. Given that the reach of AOD treatment has expanded via the online medium, in this article we examine how 'problems' are produced in online alcohol counselling encounters involving people aged 55 and over. Drawing on poststructural approaches to problematisation, we not only trace how and what 'problems' are produced, but also what effects these give rise to. We discuss three approaches to problem formulation: (1) Addiction discourses at work; (2) Moving between concerns and alcohol 'problems'; (3) Making 'problems' complex and multiple. On the basis of this analysis, we argue that online AOD counselling does not just respond to pre-existing 'AOD problems'. Rather, through the social and clinical practices of formulation at work in clinical encounters, online counselling also produces them. Thus, given a different set of circumstances, practices and relations, 'problems' might be defined or emerge differently-perhaps not as 'problems' at all or perhaps as different kinds of concerns. We conclude by highlighting the need for a critical reflexivity in AOD treatment and policy in order to open up possibilities for different ways of engaging with, and responding to, people's needs in their complexity

  12. Spreading the word: A process evaluation of a voluntary AOD prevention program.

    PubMed

    Edelen, Maria Orlando; Tucker, Joan; D'Amico, Elizabeth

    2015-06-01

    Research on voluntary after-school alcohol and other drug (AOD) prevention programs is limited. It is important to increase understanding of students' motivation to attend these types of programs and their tendency to transfer program information to peers. This paper summarizes efforts to evaluate process information for CHOICE, a voluntary after-school AOD prevention program for middle-school youth. A survey administered to 1899 students aged 10-16 in seven schools assessed: (1) why students choose to attend CHOICE (2) barriers to attendance; and (3) how program information is disseminated to non-participants. Frequencies of responses from participants and non-participants were compared. Participants were motivated by several features, most notably, the demeanor of the group leaders and enjoyable curriculum content. Barriers to attendance were primarily logistic, but results also suggest that the promotion message should more effectively emphasize that CHOICE is appropriate for everyone. The majority of students knew about CHOICE, both through advertising and conversations with friends. Non-participants' detailed reports of what they heard from friends corresponded closely with what participants reported sharing. The use of dynamic group leaders is critical to engaging students in voluntary programs. Offering the program on different days of the week or at different times (e.g., before school) may improve attendance rates. Peer networks represent a critical pathway for prevention information that can help increase program impact. These results can be used to inform modifications to existing voluntary after-school AOD prevention programs to obtain higher attendance rates and more widespread dissemination of the intervention message. © American Academy of Addiction Psychiatry.

  13. AOD trends during 2001-2010 from observations and model simulations

    NASA Astrophysics Data System (ADS)

    Pozzer, Andrea; de Meij, Alexander; Yoon, Jongmin; Astitha, Marina

    2016-04-01

    The trend of aerosol optical depth (AOD) between 2001 and 2010 is estimated globally and regionally from remote sensed observations by the MODIS (Moderate Resolution Imaging Spectroradiometer), MISR (Multi-angle Imaging SpectroRadiometer) and SeaWIFS (Sea-viewing Wide Field-of-view Sensor) satellite sensor. The resulting trends have been compared to model results from the EMAC (ECHAM5/MESSy Atmospheric Chemistry {[1]}), model. Although interannual variability is applied only to anthropogenic and biomass-burning emissions, the model is able to quantitatively reproduce the AOD trends as observed by MODIS, while some discrepancies are found when compared to MISR and SeaWIFS. An additional numerical simulation with the same model was performed, neglecting any temporal change in the emissions, i.e. with no interannual variability for any emission source. It is shown that decreasing AOD trends over the US and Europe are due to the decrease in the (anthropogenic) emissions. On contrary over the Sahara Desert and the Middle East region, the meteorological/dynamical changes in the last decade play a major role in driving the AOD trends. Further, over Southeast Asia, both meteorology and emissions changes are equally important in defining AOD trends {[2]}. Finally, decomposing the regional AOD trends into individual aerosol components reveals that the soluble components are the most dominant contributors to the total AOD, as their influence on the total AOD is enhanced by the aerosol water content. {[1]}: Jöckel, P., Kerkweg, A., Pozzer, A., Sander, R., Tost, H., Riede, H., Baumgaertner, A., Gromov, S., and Kern, B.: Development cycle 2 of the Modular Earth Submodel System (MESSy2), Geosci. Model Dev., 3, 717-752, doi:10.5194/gmd-3-717-2010, 2010. {[2]}: Pozzer, A., de Meij, A., Yoon, J., Tost, H., Georgoulias, A. K., and Astitha, M.: AOD trends during 2001-2010 from observations and model simulations, Atmos. Chem. Phys., 15, 5521-5535, doi:10.5194/acp-15-5521-2015, 2015.

  14. Spatio-temporal PM and AOD estimations over Northeast Asia during DRAGON NE-Asia campaign

    NASA Astrophysics Data System (ADS)

    Park, M.; Song, C.; Kim, J.

    2013-12-01

    Particulate matter (PM) is closely related to human health, air quality, and climate changes. It has been directly measured on the surface level. However, ground-based measurements have a limitation in spatial coverage of PM concentrations. In order to overcome this spatial limitation of ground measurements, AOD, which is considered as a proxy to PM concentration, was used in this study. AOD was first utilized to figure out the characteristics of PM and was then used to estimate the PM concentrations in Northeast Asia during the DRAGON Northeast-Asia campaign (March-May 2012), using CMAQ-estimated AOD, COMS/GOCI-retrieved AOD, and the AOD data from the DRAGON NE-Asia campaign. First of all, current emission inventories (MEIC and INTEX-B based emission inventories) were evaluated to improve CMAQ modeling results. Next, several algorithms to convert aerosol composition to AOD were evaluated using intensive measurement data from the DRAGON NE-Asia campaign. The accuracy of the CMAQ-estimated AOD was further evaluated with hourly observing GOCI-retrieved AOD. After the evaluation, CMAQ-calculated AOD was mathematically combined with GOCI-retrieved AOD via data assimilation. After this, AERONET AOD measured by the DRAGON NE-Asia campaign was again combined with the assimilated AOD from CMAQ and GOCI AODs to produce more accurate spatio-temporal AOD fields over Northeast Asia. Using several relationships between PM (PM10 and PM2.5) and AOD, the best surface-PM concentrations over the entire domain were calculated. It was then evaluated with ground-based PM2.5 measurements from the DRAGON NE-Asia campaign. A good agreement between estimated PM2.5 and measured PM2.5 over the domain was found. Finally, the PM and AOD information was used to investigate the effects of transboundary PM pollution from China to the Korean peninsula.

  15. The GEOS-5 Neural Network Retrieval (NNR) for AOD

    NASA Technical Reports Server (NTRS)

    Castellanos, Patricia; Da Silva, Arlindo

    2017-01-01

    One of the difficulties in data assimilation is the need for multi-sensor data merging that can account for temporal and spatial biases between satellite sensors. In the Goddard Earth Observing System Model Version 5 (GEOS-5) aerosol data assimilation system, a neural network retrieval (NNR) is used as a mapping between satellite observed top of the atmosphere (TOA) reflectance and AOD, which is the target variable that is assimilated in the model. By training observations of TOA reflectance from multiple sensors to map to a common AOD dataset (in this case AOD observed by the ground based Aerosol Robotic Network, AERONET), we are able to create a global, homogenous, satellite data record of AOD from MODIS observations on board the Terra and Aqua satellites. In this talk, I will present the implementation of and recent updates to the GEOS-5 NNR for MODIS collection 6 data.

  16. Alcohol oxidase protein mediated in-situ synthesized and stabilized gold nanoparticles for developing amperometric alcohol biosensor.

    PubMed

    Chinnadayyala, Somasekhar R; Santhosh, Mallesh; Singh, Naveen K; Goswami, Pranab

    2015-07-15

    A simple one step method for the alcohol oxidases (AOx) protein mediated synthesis of gold nano-particles (AuNPs) in alkaline (pH 8.5) condition with simultaneous stabilization of the nanoparticles on the AOx protein surface under native environment has been developed. The formation of the AOx conjugated AuNPs was confirmed by advanced analytical and spectroscopic techniques. The significant increase in zeta potential (ζ) value of -57mV for the synthesized AOx-AuNPs conjugate from the AOx (pI 4.5) protein (ζ, -30mV) implied good stability of the in-situ synthesized nano-conjugate. The AOx-AuNPs conjugate showed steady stability in alkaline (upto pH 8.5) and NaCl (up to 10(-1)M) solutions. The efficiency (Kcat/Km) of the AuNP conjugated AOx was increased by 18% from the free enzyme confirming the activating role of the surface stabilized AuNPs for the enzyme. The AuNPs-AOx conjugate was encapsulated with polyaniline (PANI) synthesized by oxidative polymerization of aniline using H2O2 generated in-situ from the AOx catalysed oxidation of alcohol. The PANI encapsulated AuNPs-AOx assembly was stabilized on a glassy carbon electrode (GCE) by chitosan-Nafion mixture and then utilized the fabricated bioelectrode for detection of alcohol amperometrically using H2O2 as redox indicator at +0.6V. The constructed biosensor showed high operational stability (6.3% loss after 25 measurements), wide linear detection range of 10µM-4.7mM (R(2)=0.9731), high sensitivity of 68.3±0.35µAmM(-1) and low detection limit of 7±0.027µM for ethanol. The fabricated bioelectrode was successfully used for the selective determination of alcohol in beverage samples. Copyright © 2015 Elsevier B.V. All rights reserved.

  17. Defective monocyte oxidative burst predicts infection in alcoholic hepatitis and is associated with reduced expression of NADPH oxidase.

    PubMed

    Vergis, Nikhil; Khamri, Wafa; Beale, Kylie; Sadiq, Fouzia; Aletrari, Mina O; Moore, Celia; Atkinson, Stephen R; Bernsmeier, Christine; Possamai, Lucia A; Petts, Gemma; Ryan, Jennifer M; Abeles, Robin D; James, Sarah; Foxton, Matthew; Hogan, Brian; Foster, Graham R; O'Brien, Alastair J; Ma, Yun; Shawcross, Debbie L; Wendon, Julia A; Antoniades, Charalambos G; Thursz, Mark R

    2017-03-01

    In order to explain the increased susceptibility to serious infection in alcoholic hepatitis, we evaluated monocyte phagocytosis, aberrations of associated signalling pathways and their reversibility, and whether phagocytic defects could predict subsequent infection. Monocytes were identified from blood samples of 42 patients with severe alcoholic hepatitis using monoclonal antibody to CD14. Phagocytosis and monocyte oxidative burst (MOB) were measured ex vivo using flow cytometry, luminometry and bacterial killing assays. Defects were related to the subsequent development of infection. Intracellular signalling pathways were investigated using western blotting and PCR. Interferon-γ (IFN-γ) was evaluated for its therapeutic potential in reversing phagocytic defects. Paired longitudinal samples were used to evaluate the effect of in vivo prednisolone therapy. MOB, production of superoxide and bacterial killing in response to Escherichia coli were markedly impaired in patients with alcoholic hepatitis. Pretreatment MOB predicted development of infection within two weeks with sensitivity and specificity that were superior to available clinical markers. Accordingly, defective MOB was associated with death at 28 and 90 days. Expression of the gp91 phox subunit of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase was reduced in patients with alcoholic hepatitis demonstrating defective MOB. Monocytes were refractory to IFN-γ stimulation and showed high levels of a negative regulator of cytokine signalling, suppressor of cytokine signalling-1. MOB was unaffected by 7 days in vivo prednisolone therapy. Monocyte oxidative burst and bacterial killing is impaired in alcoholic hepatitis while bacterial uptake by phagocytosis is preserved. Defective MOB is associated with reduced expression of NADPH oxidase in these patients and predicts the development of infection and death. Published by the BMJ Publishing Group Limited. For permission to use (where not already

  18. Content and Functionality of Alcohol and Other Drug Websites: Results of an Online Survey

    PubMed Central

    White, Angela; Kavanagh, David; Shandley, Kerrie; Kay-Lambkin, Frances; Proudfoot, Judith; Drennan, Judy; Connor, Jason; Baker, Amanda; Young, Ross

    2010-01-01

    Background There is a growing trend for individuals to seek health information from online sources. Alcohol and other drug (AOD) use is a significant health problem worldwide, but access and use of AOD websites is poorly understood. Objective To investigate content and functionality preferences for AOD and other health websites. Methods An anonymous online survey examined general Internet and AOD-specific usage and search behaviors, valued features of AOD and health-related websites (general and interactive website features), indicators of website trustworthiness, valued AOD website tools or functions, and treatment modality preferences. Results Surveys were obtained from 1214 drug (n = 766) and alcohol website users (n = 448) (mean age 26.2 years, range 16-70). There were no significant differences between alcohol and drug groups on demographic variables, Internet usage, indicators of website trustworthiness, or on preferences for AOD website functionality. A robust website design/navigation, open access, and validated content provision were highly valued by both groups. While attractiveness and pictures or graphics were also valued, high-cost features (videos, animations, games) were minority preferences. Almost half of respondents in both groups were unable to readily access the information they sought. Alcohol website users placed greater importance on several AOD website tools and functions than did those accessing other drug websites: online screening tools (χ²2 = 15.8, P < .001, n = 985); prevention programs (χ²2 = 27.5, P < .001, n = 981); tracking functions (χ²2 = 11.5, P = .003, n = 983); self help treatment programs (χ²2 = 8.3, P = .02, n = 984); downloadable fact sheets for friends (χ²2 = 11.6, P = .003, n = 981); or family (χ²2 = 12.7, P = .002, n = 983). The most preferred online treatment option for both the user groups was an Internet site with email therapist support. Explorations of demographic differences were also performed. While

  19. Are perceptions of social norms regarding peer alcohol and other drug use associated with personal use in Danish adolescents?

    PubMed

    Vallentin-Holbech, Lotte; Rasmussen, Birthe Marie; Stock, Christiane

    2017-12-01

    This study aims to describe norm perceptions among Danish pupils aged 13-17 years related to the prevalence of personal lifetime use of alcohol and other drugs (AODs). Further we examined if norm perceptions were associated with personal lifetime AOD use. The data were collected as baseline data in the trial The GOOD Life. A total of 2601 pupils from 42 public schools in the Region of Southern Denmark completed an online questionnaire measuring personal lifetime AOD use and personal approval of use. Additionally the perceived frequency of AOD use and approval of use among peers of their own grade were measured. Lifetime AOD outcome variables were alcohol consumption (at least one drink, being drunk and had five or more drinks on one occasion), smoking, and cannabis use. Pupils' perceptions of peer approval were significantly higher than pupils' personal approval of AOD use among adolescents for all outcomes. With the exception of cannabis use the estimated AOD prevalence among peers (median) were higher than the actual prevalence of personal lifetime use. Multilevel logistic regression models showed a significantly increased risk of personal AOD use for pupils that overestimated their peers' AOD use and also for pupils that perceived peers to approve of AOD use. The findings highlight that pupils' exaggerated perceptions regarding their peers' use and approval of AOD use are related to personal experience with AODs.

  20. Alcohol- and drug-related absenteeism: a costly problem.

    PubMed

    Roche, Ann; Pidd, Ken; Kostadinov, Victoria

    2016-06-01

    Absenteeism related to alcohol and other drug (AOD) use can place a substantial burden on businesses and society. This study estimated the cost of AOD-related absenteeism in Australia using a nationally representative dataset. A secondary analysis of the 2013 National Drug Strategy Household Survey (n=12,196) was undertaken. Two measures of AOD-related absenteeism were used: participants' self-reported absence due to AOD use (M1); and the mean difference in absence due to any illness/injury for AOD users compared to abstainers (M2). Both figures were multiplied by $267.70 (average day's wage in 2013 plus 20% on-costs) to estimate associated costs. M1 resulted in an estimation of 2.5 million days lost annually due to AOD use, at a cost of more than $680 million. M2 resulted in an estimation of almost 11.5 million days lost, at a cost of $3 billion. AOD-related absenteeism represents a significant and preventable impost upon Australian businesses. Workplaces should implement evidence-based interventions to promote healthy employee behaviour and reduce AOD-related absenteeism. © 2015 Public Health Association of Australia.

  1. The extent of problematic alcohol and other drug use within selected South African workplaces.

    PubMed

    Harker Burnhams, Nadine; Dada, Siphokazi; Linda, Betty; Myers, Bronwyn; Parry, Charles

    2013-09-30

    The use of alcohol and other drugs (AODs) in the workplace has a major impact on the health and productivity of the workforce globally. Yet information on this issue is limited in South Africa (SA). To describe the nature and extent of AOD problems in selected workplace settings in SA. Secondary data analysis was conducted on a large dataset compiled by an Employee Assistance Programme (EAP) service provider in SA. As part of an EAP risk audit, assessments for AOD-related problems were completed for each employee accessing EAP services (n=10 428) between 2005 and 2011. Data on the socio-demographic profiles, AOD use and impact of AOD use on the work performance of employees were analysed. Findings indicate that employed men are more likely to experience alcohol-related problems than women, the latter demonstrating a higher percentage of drug-related problems. The majority of referrals to the EAPs emanate from the public, industrial and financial sectors. AOD-related problems were also found to significantly impact on employee work performance. The study begins to address the knowledge gap on the extent of AOD use in SA workplaces and points to the need for further investigations into the exact nature of AOD use. In addition, the study highlights the need for intervention programmes and policies suited to the workplace.

  2. Do workplace policies work? An examination of the relationship between alcohol and other drug policies and workers' substance use.

    PubMed

    Pidd, Ken; Kostadinov, Victoria; Roche, Ann

    2016-02-01

    There is growing interest in workplace policies as a strategy to prevent or manage alcohol and other drug (AOD) problems. This study is the first to explore the prevalence and impact of AOD policies in Australian workplaces using a nationally representative dataset. A secondary analysis of the 2010 National Drug Strategy Household Survey was conducted (n=13,590). Descriptive analyses explored the prevalence of AOD policies. Multinomial and logistic regression assessed the relationship between policies and health behaviours. Workplace AOD policies were associated with reduced employee substance use. Having any AOD policy in place was associated with significantly decreased odds of high risk drinking (OR: 0.61). In terms of specific policy types, policies on 'use' and 'use plus assistance' were associated with significantly decreased odds of high risk drinking (OR: 0.64 and 0.43, respectively). 'Comprehensive' policies were associated with significantly decreased odds of drug use (OR: 0.72). AOD policies were not significantly related to absence due to AOD use, attending work under the influence, or usually consuming AOD at work. These findings provide empirical support for the value and efficacy of policies to reduce alcohol and drug problems. While basic policies on 'use' were associated with a reduction in high risk drinking, more comprehensive policies were required to impact drug use. Notably, alcohol/drug testing in isolation does not appear to be related to reduced employee substance use. Scope exists for Australian workplaces to implement effective AOD policies. This could result in considerable benefits for both individuals and workplaces. Copyright © 2015 Elsevier B.V. All rights reserved.

  3. Monthly analysis of PM ratio characteristics and its relation to AOD.

    PubMed

    Sorek-Hamer, Meytar; Broday, David M; Chatfield, Robert; Esswein, Robert; Stafoggia, Massimo; Lepeule, Johanna; Lyapustin, Alexei; Kloog, Itai

    2017-01-01

    Airborne particulate matter (PM) is derived from diverse sources-natural and anthropogenic. Climate change processes and remote sensing measurements are affected by the PM properties, which are often lumped into homogeneous size fractions that show spatiotemporal variation. Since different sources are attributed to different geographic locations and show specific spatial and temporal PM patterns, we explored the spatiotemporal characteristics of the PM 2.5 /PM 10 ratio in different areas. Furthermore, we examined the statistical relationships between AERONET aerosol optical depth (AOD) products, satellite-based AOD, and the PM ratio, as well as the specific PM size fractions. PM data from the northeastern United States, from San Joaquin Valley, CA, and from Italy, Israel, and France were analyzed, as well as the spatial and temporal co-measured AOD products obtained from the MultiAngle Implementation of Atmospheric Correction (MAIAC) algorithm. Our results suggest that when both the AERONET AOD and the AERONET fine-mode AOD are available, the AERONET AOD ratio can be a fair proxy for the ground PM ratio. Therefore, we recommend incorporating the fine-mode AERONET AOD in the calibration of MAIAC. Along with a relatively large variation in the observed PM ratio (especially in the northeastern United States), this shows the need to revisit MAIAC assumptions on aerosol microphysical properties, and perhaps their seasonal variability, which are used to generate the look-up tables and conduct aerosol retrievals. Our results call for further scrutiny of satellite-borne AOD, in particular its errors, limitations, and relation to the vertical aerosol profile and the particle size, shape, and composition distribution. This work is one step of the required analyses to gain better understanding of what the satellite-based AOD represents. The analysis results recommend incorporating the fine-mode AERONET AOD in MAIAC calibration. Specifically, they indicate the need to revisit

  4. Spatiotemporal fusion of multiple-satellite aerosol optical depth (AOD) products using Bayesian maximum entropy method

    NASA Astrophysics Data System (ADS)

    Tang, Qingxin; Bo, Yanchen; Zhu, Yuxin

    2016-04-01

    Merging multisensor aerosol optical depth (AOD) products is an effective way to produce more spatiotemporally complete and accurate AOD products. A spatiotemporal statistical data fusion framework based on a Bayesian maximum entropy (BME) method was developed for merging satellite AOD products in East Asia. The advantages of the presented merging framework are that it not only utilizes the spatiotemporal autocorrelations but also explicitly incorporates the uncertainties of the AOD products being merged. The satellite AOD products used for merging are the Moderate Resolution Imaging Spectroradiometer (MODIS) Collection 5.1 Level-2 AOD products (MOD04_L2) and the Sea-viewing Wide Field-of-view Sensor (SeaWiFS) Deep Blue Level 2 AOD products (SWDB_L2). The results show that the average completeness of the merged AOD data is 95.2%,which is significantly superior to the completeness of MOD04_L2 (22.9%) and SWDB_L2 (20.2%). By comparing the merged AOD to the Aerosol Robotic Network AOD records, the results show that the correlation coefficient (0.75), root-mean-square error (0.29), and mean bias (0.068) of the merged AOD are close to those (the correlation coefficient (0.82), root-mean-square error (0.19), and mean bias (0.059)) of the MODIS AOD. In the regions where both MODIS and SeaWiFS have valid observations, the accuracy of the merged AOD is higher than those of MODIS and SeaWiFS AODs. Even in regions where both MODIS and SeaWiFS AODs are missing, the accuracy of the merged AOD is also close to the accuracy of the regions where both MODIS and SeaWiFS have valid observations.

  5. Survey of attitudes and behaviors toward alcohol and other drug use in allied health and physician assistant students.

    PubMed

    Baldwin, Jeffrey N; Davis-Hall, R Ellen; DeSimone, Edward M; Scott, David M; Agrawal, Sangeeta; Reardon, Thomas P

    2008-01-01

    Attitudes and behaviors toward alcohol and other drug (AOD) use were assessed among a subgroup of allied health (AH) and physician assistant (PA) students within two university-based health professions educational institutions in a midwestern state in 1999. AH/PA student response was 423 (77.5%); this included occupational therapy (n = 81; 60.4%), physical therapy [PT] (n = 222; 98.7%), PA (n = 68; 85.0%), and other AH programs (n = 52; 48.1%). In this AH/PA group, a family history of alcohol-related problems was reported by 40.1% and drug-related problems by 11.3%, with 42.5% of respondents reporting one or both. Such histories of family alcohol and drug problems were reported by 47.8% or 18.5% of PA students, respectively. Past-year alcohol, tobacco, and marijuana use were reported, respectively, by 88.1%, 26.1%, and 6.7%. The highest percentage use of marijuana was reported by PT students (9.0%). Past-year blackouts were reported by 21.9%, driving after three or more drinks or any drug use by 49.3%, attendance at class or work under the influence of AOD by 7.8%, patient care under the influence of AOD by 3.3%, lowered grades or job evaluations from AOD use by 5.0%, and AOD-related legal charges by 4.5%. "Heavy drinking" (defined as consumption of five or more drinks per occasion during the past 2 weeks) was reported by 33.2%. Inadequate AOD education was reported by 55.1%, with 63.6% of PT respondents so reporting. AH and PA educational systems should proactively address student AOD prevention, education, and assistance needs.

  6. Toxicity assessment and geochemical model of chromium leaching from AOD slag.

    PubMed

    Liu, Bao; Li, Junguo; Zeng, Yanan; Wang, Ziming

    2016-02-01

    AOD (Argon Oxygen Decarburization) slag is a by-product of the stainless steel refining process. The leaching toxicity of chromium from AOD slag cannot be ignored in the recycling process of the AOD slag. To assess the leaching toxicity of the AOD slag, batch leaching tests have been performed. PHREEQC simulations combined with FactSage were carried out based on the detailed mineralogical analysis and petrophysical data. Moreover, Pourbaix diagram of the Cr-H2O system was protracted by HSC 5.0 software to explore the chromium speciation in leachates. It was found that AOD slag leachate is an alkaline and reductive solution. The Pourbaix diagram of the Cr-H2O system indicated that trivalent chromium, such as Cr(OH)4(-), is the major chromium species in the experimental Eh-pH region considered. However, toxic hexavalent chromium was released with maximum concentrations of 30 µg L(-1) and 18 µg L(-1) at L/S 10 and 100, respectively, during the earlier leaching stage. It concluded that the AOD slag possessed a certain leaching toxicity. After 10 d of leaching, trivalent chromium was the dominant species in the leachates, which corresponded to the results of PHREEQC simulation. Leaching toxicity of AOD slag is based on the chromium speciation and its transformation. Great attention should be focused on such factors as aging, crystal form of chromium-enriched minerals, and electrochemical characteristics of the leachates. Copyright © 2015 Elsevier Ltd. All rights reserved.

  7. Alcohol availability and neighborhood poverty and their relationship to binge drinking and related problems among drinkers in committed relationships

    PubMed Central

    McKinney, Christy M.; Chartier, Karen G.; Caetano, Raul; Harris, T. Robert

    2012-01-01

    The authors examined the relationship of alcohol outlet density (AOD) and neighborhood poverty with binge drinking and alcohol-related problems among drinkers in married and cohabitating relationships and assessed whether these associations differed across sex. A U.S. national population couples survey was linked to U.S. Census data on AOD and neighborhood poverty. The 1,784 current drinkers in the survey reported on their binge drinking, alcohol-related problems, and other covariates. AOD was defined as the number of alcohol outlets per 10,000 persons and was obtained at the zip code level. Neighborhood poverty was as having a low (<20%) or high (≥20%) proportion of residents living in poverty at the census tract level. We used logistic regression for survey data to estimate odds ratios and 95% confidence intervals and tested for differences of associations by sex. Associations of neighborhood poverty with binge drinking were stronger for male than for female drinkers. The association of neighborhood poverty with alcohol-related problems was also stronger for men than for women. We observed no relationships between AOD and binge drinking or alcohol-related problems in this couples survey. Efforts to reduce binge drinking or alcohol-related problems among partners in committed relationships may have the greatest impact if targeted to male drinkers living in high-poverty neighborhoods. Binge drinking and alcohol-related problems, as well as residence in an impoverished neighborhood are risk factors for intimate partner violence (IPV) and other relationship conflicts. PMID:22890980

  8. Alcohol availability and neighborhood poverty and their relationship to binge drinking and related problems among drinkers in committed relationships.

    PubMed

    McKinney, Christy M; Chartier, Karen G; Caetano, Raul; Harris, T Robert

    2012-09-01

    The authors examined the relationship of alcohol outlet density (AOD) and neighborhood poverty with binge drinking and alcohol-related problems among drinkers in married and cohabitating relationships and assessed whether these associations differed across sex. A U.S. national population couples survey was linked to U.S. Census data on AOD and neighborhood poverty. The 1,784 current drinkers in the survey reported on their binge drinking, alcohol-related problems, and other covariates. AOD was defined as the number of alcohol outlets per 10,000 persons and was obtained at the zip code level. Neighborhood poverty was defined as having a low (<20%) or high (≥20%) proportion of residents living in poverty at the census tract level. We used logistic regression for survey data to estimate odds ratios and 95% confidence intervals and tested for differences of associations by sex. Associations of neighborhood poverty with binge drinking were stronger for male than for female drinkers. The association of neighborhood poverty with alcohol-related problems was also stronger for men than for women. We observed no relationships between AOD and binge drinking or alcohol-related problems in this couples survey. Efforts to reduce binge drinking or alcohol-related problems among partners in committed relationships may have the greatest impact if targeted to male drinkers living in high-poverty neighborhoods. Binge drinking and alcohol-related problems, as well as residence in an impoverished neighborhood are risk factors for intimate partner violence (IPV) and other relationship conflicts.

  9. Mothers' alcohol and other drug problems and their punitiveness toward their children.

    PubMed

    Miller, B A; Smyth, N J; Mudar, P J

    1999-09-01

    The relationship between mothers' alcohol or other drug (AOD) problems and their punitiveness toward their children is examined in this study. Women (N = 170) were recruited from five different sources, including clinical interventions and the community. Women's punitiveness toward their children was assessed by three different maternal self-report measures: Conflict Tactics Scale, Parental Punitiveness Scale and Child Abuse Potential Inventory. Women's histories of AOD problems, violent victimization and hostility were assessed using standardized assessment tools. Women were categorized by their AOD problems as follows: current problem (22.4%), past only problem (44.7%) or no problems (32.9%). Results indicate that mothers with current or past AOD problems are more punitive toward their children, controlling for potentially confounding demographic factors. Mothers' histories of partner violence and parental violence also predicted higher levels of mother-to-child punitiveness. However, histories of childhood sexual abuse predicted lower levels of mothers' punitiveness. Hostility served as a predictor of mothers' punitiveness and moderated some of the relationships between their AOD problems and their punitiveness. Findings suggest that women with AOD problems are more likely to be punitive toward their children, but other factors may also contribute to this risk. Treatment and prevention interventions need to be developed to decrease the risk of violent victimization among children whose mothers have alcohol and other drug problems.

  10. a Thtee-Dimensional Variational Assimilation Scheme for Satellite Aod

    NASA Astrophysics Data System (ADS)

    Liang, Y.; Zang, Z.; You, W.

    2018-04-01

    A three-dimensional variational data assimilation scheme is designed for satellite AOD based on the IMPROVE (Interagency Monitoring of Protected Visual Environments) equation. The observation operator that simulates AOD from the control variables is established by the IMPROVE equation. All of the 16 control variables in the assimilation scheme are the mass concentrations of aerosol species from the Model for Simulation Aerosol Interactions and Chemistry scheme, so as to take advantage of this scheme in providing comprehensive analyses of species concentrations and size distributions as well as be calculating efficiently. The assimilation scheme can save computational resources as the IMPROVE equation is a quadratic equation. A single-point observation experiment shows that the information from the single-point AOD is effectively spread horizontally and vertically.

  11. The effects of purchasing alcohol and marijuana among adolescents at-risk for future substance use.

    PubMed

    Osilla, Karen Chan; Pedersen, Eric R; Ewing, Brett A; Miles, Jeremy N V; Ramchand, Rajeev; D'Amico, Elizabeth J

    2014-09-18

    Among high-risk youth, those who may be at increased risk for adverse alcohol and other drug (AOD) use outcomes may benefit from targeted prevention efforts; how youth acquire AOD may provide an objective means of identifying youth at elevated risk. We assessed how youth acquired alcohol and marijuana (purchasing vs. other means), demographics, AOD behaviors/consequences, and environment among adolescents referred to a diversion program called Teen Court (N = 180) at two time points (prior to the program and 180 days from baseline). Participants were predominantly White and Hispanic/Latino(a). In cross-sectional analyses among alcohol and marijuana users, purchasing marijuana was associated with more frequent marijuana use and consequences, time spent around teens who use marijuana, higher likelihood of substance use disorders, and lower resistance self-efficacy compared to non-purchasers. Teens who purchased both alcohol and marijuana experienced similar outcomes to those who purchased only marijuana, and also reported more frequent and higher quantity of drinking, greater alcohol-related consequences, time spent around teens who use other drugs, and prescription drug misuse. Longitudinally, purchasing alcohol and marijuana at baseline was associated with more frequent and higher quantity of drinking compared to non-purchasers at follow-up. Marijuana only purchasers had a greater likelihood of substance use disorders at follow-up compared to non-purchasers. In an era where drinking is commonplace and attitudes towards marijuana use are becoming more tolerant, it is essential to evaluate how accessibility to AOD and subsequent purchasing behaviors affect youth consumption and intervene accordingly to prevent future consequences.

  12. A reagentless amperometric biosensor for alcohol detection in column liquid chromatography based on co-immobilized peroxidase and alcohol oxidase in carbon paste.

    PubMed

    Johansson, K; Jönsson-Pettersson, G; Gorton, L; Marko-Varga, G; Csöregi, E

    1993-12-01

    A reagentless carbon paste electrode chemically modified with covalently bound alcohol oxidase and horse-radish peroxidase was examined as a selective sensor in flow injection and column liquid chromatography. A combination of carbodiimide, glutaraldehyde, and polyethyleneimine was used for immobilizing the enzymes in the paste. The surface of the electrodes was protected by first forming a layer of electropolymerized ortho-phenylenediamine followed by deposition of a cation exchange membrane (Eastman AQ 29D). The electrodes were used for detection of hydrogen peroxide, methanol, ethanol, propanol, isopropanol, and butanol. Preliminary investigations of the use of this sensor for bioprocess control are reported.

  13. Alcohol and other psychoactive drugs in trauma patients aged 10–14 years

    PubMed Central

    Li, G.; Chanmugam, A.; Rothman, R.; DiScala, C.; Paidas, C.; Kelen, G.

    1999-01-01

    Objective—To examine the prevalence of alcohol and/or other psychoactive drugs, such as marijuana and cocaine (AODs), involved in preteen trauma patients. Methods—Toxicological testing results were analyzed for 1356 trauma patients aged 10–14 years recorded in the National Pediatric Trauma Registry for the years 1990–95. Results—Of the 1356 patients who received toxicological screening at the time of admission, 116 (9%) were positive for AODs. AOD involvement increased with age. Patients with pre-existing mental disorders were nearly three times as likely as other patients to be AOD positive (23% v 8%, p<0.01). AOD involvement was more prevalent in intentional injuries and in injuries that occurred at home. Conclusions—AODs in preteen trauma are of valid concern, in particular among patients with mental disorders or intentional injuries. The role of AODs in childhood injuries needs to be further examined using standard screening instruments and representative study samples. PMID:10385826

  14. Program Administrator's Handbook. Strategies for Preventing Alcohol and Other Drug Problems. The College Series.

    ERIC Educational Resources Information Center

    CSR, Inc., Washington, DC.

    This handbook is for administrators of programs in higher education settings which deal with alcohol and other drug (AOD) related problems. Chapter 1, "Defining the Problem, Issues, and Trends" examines the problem from various perspectives and presents the latest statistics on the extent of AOD use on campuses, specific problems affecting…

  15. a New Algorithm for the Aod Inversion from Noaa/avhrr Data

    NASA Astrophysics Data System (ADS)

    Sun, L.; Li, R.; Yu, H.

    2018-04-01

    The advanced very high resolution radiometer (AVHRR) data from the National Oceanic and Atmospheric Administration satellite is one of the earliest data applied in aerosol research. The dense dark vegetation (DDV) algorithm is a popular method for the present land aerosol retrieval. One of the most crucial steps in the DDV algorithm with AVHRR data is estimating the land surface reflectance (LSR). However, LSR cannot be easily estimated because of the lack of a 2.13 μm band. In this article, the moderate resolution imaging spectroradiometer (MODIS) vegetation index product (MYD13) is introduced to support the estimation of AVHRR LSR. The relationship between MODIS NDVI and the AVHRR LSR of the visible band is analysed to retrieve aerosol optical depth (AOD) from AVHRR data. Retrieval experiments are carried out in mid-eastern America. The AOD data from AErosol RObotic NETwork (AERONET) measurements are used to evaluate the aerosol retrieval from AVHRR data, the results indicate that about 74 % of the retrieved AOD are within the expected error range of ±(0.05 + 0.2), and a cross comparison of the AOD retrieval results with the MODIS aerosol product (MYD04) shows that the AOD datasets have a similar spatial distribution.

  16. Community control of alcohol and drug risk environments: the California experience.

    PubMed

    Wittman, Friedner D

    2007-01-01

    This article describes California community (city, county) uses of local powers and resources to prevent alcohol and other drug (AOD) problems by managing AOD risk environments in retail, public, and social domains. The article presents a promising framework used in several counties dedicated to community environment approaches to prevention. A case example of its application is provided. The framework has developed locally since the mid 1980s through AOD prevention demonstration grant research, policy advocacy, and state support for local initiatives. Data for this article come from the author's experience observing and consulting with California cities and counties over 25 years.

  17. Environmental Management: An Approach to Alcohol and Other Drug Prevention. Prevention Updates

    ERIC Educational Resources Information Center

    Higher Education Center for Alcohol and Other Drug Abuse and Violence Prevention, 2002

    2002-01-01

    Most campus alcohol and other drug (AOD) programs include prevention, intervention, and treatment services designed to address individual students' knowledge of the consequences of alcohol and other drug use, to improve their skills in resisting such behavior, or to address existing problematic use of or addiction to alcohol or other drugs.…

  18. Maximum availability and mineralogical control of chromium released from AOD slag.

    PubMed

    Li, Junguo; Liu, Bao; Zeng, Yanan; Wang, Ziming; Gao, Zhiyuan

    2017-03-01

    AOD (argon oxygen decarburization) slag is the by-product in the stainless steel refining process. Chromium existing in AOD slag can leach out and probably poses a serious threat to the environment. To assess the leaching toxicity of chromium released from AOD slag, the temperature-dependent maximum availability leaching test was performed. To determine the controlling mineralogical phases of chromium released from AOD slag, a Visual MINTEQ simulation was established based on Vminteq30 and the FactSage 7.0 database. The leaching tests indicated that the leaching availability of chromium was slight and mainly consisted of trivalent chromium. Aging of AOD slag under the atmosphere can oxidize trivalent chromium to hexavalent chromium, which could be leached out by rainwater. According to the simulation, the chromium concentration in leachates was controlled by the freely soluble pseudo-binary phases in the pH = 7.0 leaching process and controlled by the Cr 2 O 3 phase in the pH = 4.0 leaching process. Chromium concentrations were underestimated when the controlling phases were determined to be FeCr 2 O 4 and MgCr 2 O 4 . Facilitating the generation of the insoluble spinel-like phases during the cooling and disposal process of the molten slag could be an effective approach to decreasing the leaching concentration of chromium and its environmental risk.

  19. Monoamine oxidase-A polymorphisms might modify the association between the dopamine D2 receptor gene and alcohol dependence.

    PubMed

    Huang, San-Yuan; Lin, Wei-Wen; Wan, Fang-Jung; Chang, Ai-Ju; Ko, Huei-Chen; Wang, Tso-Jen; Wu, Pei-Lin; Lu, Ru-Band

    2007-05-01

    Low monoamine oxidase (MAO) activity and the neurotransmitter dopamine are 2 important factors in the development of alcohol dependence. MAO is an important enzyme associated with the metabolism of biogenic amines. Therefore, the present study investigates whether the association between the dopamine D2 receptor (DRD2) gene and alcoholism is affected by different polymorphisms of the MAO type A (MAOA) gene. A total of 427 Han Chinese men in Taiwan (201 control subjects and 226 with alcoholism) were recruited for the study. Of the subjects with alcoholism, 108 had pure alcohol dependence (ALC) and 118 had both alcohol dependence and anxiety, depression or both (ANX/DEP ALC). All subjects were assessed with the Chinese Version of the Modified Schedule of Affective Disorders and Schizophrenia-Lifetime. Alcohol dependence, anxiety and major depressive disorders were diagnosed according to Diagnostic and Statistical Manual of Mental Disorders, fourth edition criteria. The genetic variant of the DRD2 gene was only associated with the ANX/DEP ALC phenotype, and the genetic variant of the MAOA gene was associated with pure ALC. Subjects carrying the MAOA 3-repeat allele and genotype A1/A1 of the DRD2 were 3.48 times (95% confidence interval = 1.47-8.25) more likely to be ANX/DEP ALC than the subjects carrying the MAOA 3-repeat allele and DRD2 A2/A2 genotype. The MAOA gene may modify the association between the DRD2 gene and ANX/DEP ALC phenotype.

  20. Protein dynamics promote hydride tunnelling in substrate oxidation by aryl-alcohol oxidase.

    PubMed

    Carro, Juan; Martínez-Júlvez, Marta; Medina, Milagros; Martínez, Angel T; Ferreira, Patricia

    2017-11-01

    The temperature dependence of hydride transfer from the substrate to the N5 of the FAD cofactor during the reductive half-reaction of Pleurotus eryngii aryl-alcohol oxidase (AAO) is assessed here. Kinetic isotope effects on both the pre-steady state reduction of the enzyme and its steady-state kinetics, with differently deuterated substrates, suggest an environmentally-coupled quantum-mechanical tunnelling process. Moreover, those kinetic data, along with the crystallographic structure of the enzyme in complex with a substrate analogue, indicate that AAO shows a pre-organized active site that would only require the approaching of the hydride donor and acceptor for the tunnelled transfer to take place. Modification of the enzyme's active-site architecture by replacement of Tyr92, a residue establishing hydrophobic interactions with the substrate analogue in the crystal structure, in the Y92F, Y92L and Y92W variants resulted in different temperature dependence patterns that indicated a role of this residue in modulating the transfer reaction.

  1. Cognitive, Behavioral, and Environmental Factors in Adolescents' Alcohol and Other Drug Use: A Developmental Perspective.

    ERIC Educational Resources Information Center

    Jenkins, Jeanne E.; And Others

    This paper reports the preliminary findings of a county-wide study of alcohol and other drug (AOD) behaviors and influences among 2,229 randomly selected students in grade 8, 10, and 12. The discussion focuses on behavioral, cognitive, and environmental predictors of AOD behaviors. The paper highlights a refined study of the influence of peers,…

  2. Employee assistance program services for alcohol and other drug problems: implications for increased identification and engagement in treatment.

    PubMed

    Jacobson, Jodi M; Sacco, Paul

    2012-01-01

    Fourteen million U.S. workers meet the diagnostic criteria for substance dependence, costing millions in lost productivity. Prior research suggests that employees who follow through with their Employee Assistance Program's (EAP) recommendations are more likely to participate and remain engaged in alcohol and other drug (AOD) treatment programs. This study identified rates of lifetime EAP service use for AOD problems and compared adults who reported using EAP services for AOD problems with those who used services other than EAP. Researchers analyzed a subset of participants from the National Epidemiologic Survey of Alcohol and Related Conditions who reported having received help for an AOD problem (NESARC, 2001-2002). Statistical analyses tested for differences in sociodemographic variables, lifetime mental health and substance abuse disorders, and health disability between EAP services users and users of other types of services. Among adults who sought services for AOD problems (n= 2,272), 7.58% (n= 166) reported using EAP services for these problems at some point during their lives. Major depressive disorder (lifetime), a drug use disorder (lifetime), and Black race/ethnicity were associated with a greater likelihood that someone would seek EAP services for help with their AOD problem. Results provide a foundation for researchers to understand who uses EAP services for AOD problems. Health and mental health professionals should increase their knowledge of EAP services to improve continuity of care for employees with AOD problems. EAPs are in a unique position to reach out to vulnerable employees in the workplace and engage them in treatment. Copyright © American Academy of Addiction Psychiatry.

  3. Evaluation of VIIRS, GOCI, and MODIS Collection 6 AOD Retrievals Against Ground Sunphotometer Observations Over East Asia

    NASA Technical Reports Server (NTRS)

    Xiao, Q.; Zhang, H.; Choi, M.; Li, S.; Kondragunta, S.; Kim, J.; Holben, B.; Levy, R. C.; Liu, Y.

    2016-01-01

    Persistent high aerosol loadings together with extremely high population densities have raised serious air quality and public health concerns in many urban centers in East Asia. However, ground-based air quality monitoring is relatively limited in this area. Recently, satellite-retrieved Aerosol Optical Depth (AOD) at high resolution has become a powerful tool to characterize aerosol patterns in space and time. Using ground AOD observations from the Aerosol Robotic Network (AERONET) and the Distributed Regional Aerosol Gridded Observation Networks (DRAGON)-Asia Campaign, as well as from handheld sunphotometers, we evaluated emerging aerosol products from the Visible Infrared Imaging Radiometer Suite (VIIRS) aboard the Suomi National Polar-orbiting Partnership (S-NPP), the Geostationary Ocean Color Imager (GOCI) aboard the Communication, Ocean, and Meteorology Satellite (COMS), and Terra and Aqua Moderate Resolution Imaging Spectroradiometer (MODIS) (Collection 6) in East Asia in 2012 and 2013. In the case study in Beijing, when compared with AOD observations from handheld sunphotometers, 51% of VIIRS Environmental Data Record (EDR) AOD, 37% of GOCI AOD, 33% of VIIRS Intermediate Product (IP) AOD, 26% of Terra MODIS C6 3km AOD, and 16% of Aqua MODIS C6 3km AOD fell within the reference expected error (EE) envelope (+/-0.05/+/- 0.15 AOD). Comparing against AERONET AOD over the JapanSouth Korea region, 64% of EDR, 37% of IP, 61% of GOCI, 39% of Terra MODIS, and 56% of Aqua MODIS C6 3km AOD fell within the EE. In general, satellite aerosol products performed better in tracking the day-to-day variability than tracking the spatial variability at high resolutions. The VIIRS EDR and GOCI products provided the most accurate AOD retrievals, while VIIRS IP and MODIS C6 3km products had positive biases.

  4. Evaluation of VIIRS, GOCI, and MODIS Collection 6 AOD retrievals against ground sunphotometer observations over East Asia

    NASA Astrophysics Data System (ADS)

    Xiao, Q.; Zhang, H.; Choi, M.; Li, S.; Kondragunta, S.; Kim, J.; Holben, B.; Levy, R. C.; Liu, Y.

    2016-02-01

    Persistent high aerosol loadings together with extremely high population densities have raised serious air quality and public health concerns in many urban centers in East Asia. However, ground-based air quality monitoring is relatively limited in this area. Recently, satellite-retrieved Aerosol Optical Depth (AOD) at high resolution has become a powerful tool to characterize aerosol patterns in space and time. Using ground AOD observations from the Aerosol Robotic Network (AERONET) and the Distributed Regional Aerosol Gridded Observation Networks (DRAGON)-Asia Campaign, as well as from handheld sunphotometers, we evaluated emerging aerosol products from the Visible Infrared Imaging Radiometer Suite (VIIRS) aboard the Suomi National Polar-orbiting Partnership (S-NPP), the Geostationary Ocean Color Imager (GOCI) aboard the Communication, Ocean, and Meteorology Satellite (COMS), and Terra and Aqua Moderate Resolution Imaging Spectroradiometer (MODIS) (Collection 6) in East Asia in 2012 and 2013. In the case study in Beijing, when compared with AOD observations from handheld sunphotometers, 51 % of VIIRS Environmental Data Record (EDR) AOD, 37 % of GOCI AOD, 33 % of VIIRS Intermediate Product (IP) AOD, 26 % of Terra MODIS C6 3 km AOD, and 16 % of Aqua MODIS C6 3 km AOD fell within the reference expected error (EE) envelope (±0.05 ± 0.15 AOD). Comparing against AERONET AOD over the Japan-South Korea region, 64 % of EDR, 37 % of IP, 61 % of GOCI, 39 % of Terra MODIS, and 56 % of Aqua MODIS C6 3 km AOD fell within the EE. In general, satellite aerosol products performed better in tracking the day-to-day variability than tracking the spatial variability at high resolutions. The VIIRS EDR and GOCI products provided the most accurate AOD retrievals, while VIIRS IP and MODIS C6 3 km products had positive biases.

  5. Optical bio-sniffer for ethanol vapor using an oxygen-sensitive optical fiber.

    PubMed

    Mitsubayashi, Kohji; Kon, Takuo; Hashimoto, Yuki

    2003-11-30

    An optical bio-sniffer for ethanol was constructed by immobilizing alcohol oxidase (AOD) onto a tip of a fiber optic oxygen sensor with a tube-ring, using an oxygen sensitive ruthenium organic complex (excitation, 470 nm; fluorescent, 600 nm). A reaction unit for circulating buffer solution was applied to the tip of the device. After the experiment in the liquid phase, the sniffer-device was applied for gas analysis using a gas flow measurement system with a gas generator. The optical device was applied to detect the oxygen consumption induced by AOD enzymatic reaction with alcohol application. The sensor in the liquid phase was used to measure ethanol solution from 0.50 to 9.09 mmol/l. Then, the bio-sniffer was calibrated against ethanol vapor from 0.71 to 51.49 ppm with good gas-selectivity based on the AOD substrate specificity. The bio-sniffer with the reaction unit was also used to monitor the concentration change of gaseous ethanol by rinsing and cleaning the fiber tip and the enzyme membrane with buffer solution.

  6. Alcohol and other drug problems in Australia: the urgent need for nurse education.

    PubMed

    de Crespigny, C

    1996-07-01

    The economic, social and health costs associated with alcohol and other drug use, including medicines, impact on the Australian community profoundly. Many Australians use alcohol and other drugs (AODs) on a regular basis and a significant number experience problems at some time. There are about 25,000 drug-related deaths annually, the majority of which are related to alcohol and tobacco use. Many young people die or are injured as a direct result of alcohol intoxication, accidental overdose and related problems, and significant numbers of elderly people are hospitalised, permanently incapacitated or suffer avoidable trauma through adverse drug reactions and side-effects of medications. It is estimated that at least 25 per cent of acute hospital beds hold patients with alcohol-related problems directly associated with the medical diagnosis, and there are comparable numbers of people needing social and other forms of assistance at some time. Despite the preventative nature of many AOD problems and the call for harm reduction, timely intervention and support for the thousands of Australians attending hospitals, community health and mental health services, nurses are yet to effectively assist the community in preventing and responding to the range of AOD problems. Nursing policy, guidelines and clinical expertise are required to assist the community in addressing this issue. Undergraduate, postgraduate and continuing education have a vital role to play in providing the profession with the knowledge, skills and research base to meet this challenge.

  7. Analysis of influential factors for the relationship between PM2.5 and AOD in Beijing

    NASA Astrophysics Data System (ADS)

    Zheng, Caiwang; Zhao, Chuanfeng; Zhu, Yannian; Wang, Yang; Shi, Xiaoqin; Wu, Xiaolin; Chen, Tianmeng; Wu, Fang; Qiu, Yanmei

    2017-11-01

    The relationship between aerosol optical depth (AOD) and PM2.5 is often investigated in order to obtain surface PM2.5 from satellite observation of AOD with a broad area coverage. However, various factors could affect the AOD-PM2.5 regressions. Using both ground and satellite observations in Beijing from 2011 to 2015, this study analyzes the influential factors including the aerosol type, relative humidity (RH), planetary boundary layer height (PBLH), wind speed and direction, and the vertical structure of aerosol distribution. The ratio of PM2.5 to AOD, which is defined as η, and the square of their correlation coefficient (R2) have been examined. It shows that η varies from 54.32 to 183.14, 87.32 to 104.79, 95.13 to 163.52, and 1.23 to 235.08 µg m-3 with aerosol type in spring, summer, fall, and winter, respectively. η is smaller for scattering-dominant aerosols than for absorbing-dominant aerosols, and smaller for coarse-mode aerosols than for fine-mode aerosols. Both RH and PBLH affect the η value significantly. The higher the RH, the smaller the η, and the higher the PBLH, the smaller the η. For AOD and PM2.5 data with the correction of RH and PBLH compared to those without, R2 of monthly averaged PM2.5 and AOD at 14:00 LT increases from 0.63 to 0.76, and R2 of multi-year averaged PM2.5 and AOD by time of day increases from 0.01 to 0.93, 0.24 to 0.84, 0.85 to 0.91, and 0.84 to 0.93 in four seasons respectively. Wind direction is a key factor for the transport and spatial-temporal distribution of aerosols originated from different sources with distinctive physicochemical characteristics. Similar to the variation in AOD and PM2.5, η also decreases with the increasing surface wind speed, indicating that the contribution of surface PM2.5 concentrations to AOD decreases with surface wind speed. The vertical structure of aerosol exhibits a remarkable change with seasons, with most particles concentrated within about 500 m in summer and within 150 m in winter

  8. Analysis of Influential Factors for the Relationship between PM2.5 and AOD in Beijing

    NASA Astrophysics Data System (ADS)

    Zheng, Caiwang; Zhao, Chuanfeng

    2017-04-01

    Relationship between aerosol optical depth (AOD) and PM2.5 is often investigated in order to obtain surface PM2.5 from satellite observation of AOD with a broad area coverage. However, various factors could affect the AOD-PM2.5 regressions. Using both ground and satellite observations in Beijing from 2011 to 2015, this study analyzes the influential factors including aerosol type, relative humidity (RH), atmospheric boundary layer height (PBLH), wind speed and direction, and the vertical structure of aerosol distribution. The ratio of PM2.5 to AOD, which is defined as η, and the square of their correlation coefficient (R2) have been examined. It shows that η varies from 54.32 to 183.14, 87.32 to 104.79, 95.13 to 163.52 and 1.23 to 235.08 μg/m3 with aerosol type in four seasons respectively. η is smaller for scattering-dominant aerosols than for absorbing-dominant aerosols, and smaller for coarse mode aerosols than for fine mode aerosols. Both RH and PBLH affect the η value significantly. The higher the RH, the larger the η, and the higher the PBLH, the smaller the η. For AOD and PM2.5 data with the correction of RH and PBLH compared to those without, R2 of monthly averaged PM2.5and AOD at 14:00 LT increases from 0.63 to 0.76, and R2 of multi-year averaged PM2.5and AOD by time of day increases from 0.1 to 0.93, 0.24 to 0.84, 0.85 to 0.91 and 0.84 to 0.93 in four seasons respectively. Wind direction is a key factor to the transport and spatial-temporal distribution of aerosols originated from different sources with distinctive physicochemical characteristics. Similar to the variation of AOD and PM2.5, η also decreases with the increasing surface wind speed, indicating that the contribution of surface PM2.5 concentrations to AOD decreases with surface wind speed. The vertical structure of aerosol exhibits a remarkable change with seasons, with most particles concentrated within about 500 m in summer and within 150 m in winter. Compared to the AOD of the whole

  9. The substrate oxidation mechanism of pyranose 2-oxidase and other related enzymes in the glucose-methanol-choline superfamily.

    PubMed

    Wongnate, Thanyaporn; Chaiyen, Pimchai

    2013-07-01

    Enzymes in the glucose-methanol-choline (GMC) oxidoreductase superfamily catalyze the oxidation of an alcohol moiety to the corresponding aldehyde. In this review, the current understanding of the sugar oxidation mechanism in the reaction of pyranose 2-oxidase (P2O) is highlighted and compared with that of other enzymes in the GMC family for which structural and mechanistic information is available, including glucose oxidase, choline oxidase, cholesterol oxidase, cellobiose dehydrogenase, aryl-alcohol oxidase, and pyridoxine 4-oxidase. Other enzymes in the family that have been newly discovered or for which less information is available are also discussed. A large primary kinetic isotope effect was observed for the flavin reduction when 2-d-D-glucose was used as a substrate, but no solvent kinetic isotope effect was detected for the flavin reduction step. The reaction of P2O is consistent with a hydride transfer mechanism in which there is stepwise formation of d-glucose alkoxide prior to the hydride transfer. Site-directed mutagenesis of P2O and pH-dependence studies indicated that His548 is a catalytic base that facilitates the deprotonation of C2-OH in D-glucose. This finding agrees with the current mechanistic model for aryl-alcohol oxidase, glucose oxidase, cellobiose dehydrogenase, methanol oxidase, and pyridoxine 4-oxidase, but is different from that of cholesterol oxidase and choline oxidase. Although all of the GMC enzymes share similar structural folding and use the hydride transfer mechanism for flavin reduction, they appear to have subtle differences in the fine-tuned details of how they catalyze substrate oxidation. © 2013 The Authors Journal compilation © 2013 FEBS.

  10. The observation-based relationships between PM2.5 and AOD over China

    NASA Astrophysics Data System (ADS)

    Xin, Jinyuan; Gong, Chongshui; Liu, Zirui; Cong, Zhiyuan; Gao, Wenkang; Song, Tao; Pan, Yuepeng; Sun, Yang; Ji, Dongsheng; Wang, Lili; Tang, Guiqian; Wang, Yuesi

    2016-09-01

    This is the first investigation of the generalized linear regressions of PM2.5 and aerosol optical depth (AOD) with the Campaign on atmospheric Aerosol Research-China network over the large high-concentration aerosol region during the period from 2012 to 2013. The map of the PM2.5 and AOD levels showed large spatial differences in the aerosol concentrations and aerosol optical properties over China. The ranges of the annual mean PM2.5 and AOD were 10-117 µg/m3 and 0.12-1.11 from the clean regions to seriously polluted regions, from the almost "arctic" and the Tibetan Plateau to tropical environments. There were significant spatial agreements and correlations between the PM2.5 and AOD. However, the linear regression functions (PM2.5 = A*AOD + B) exhibited large differences in different regions and seasons. The slopes (A) were from 13 to 90, the intercepts (B) were from 0.8 to 33.3, and the correlation coefficients (R2) ranged from 0.06 to 0.75. The slopes (A) were much higher in the north (41-99) than in the south (13-64) because the extinction efficiency of hygroscopic aerosol was rapidly increasing with the increasing humidity from the dry north to the humid south. Meanwhile, the intercepts (B) were generally lower, and the correlation coefficients (R2) were much higher in the dry north than in the humid south. There was high consistency of AOD versus PM2.5 for all sites in three ranges of the atmospheric column precipitable water vapor (PWV). The segmented linear regression functions were y = 84.66x + 9.85 (PWV < 1.0), y = 69.47x + 11.87 (1.0 < PWV < 2.5), and y = 52.37x + 8.59 (PWV > 2.5). The correlation coefficients (R2) were high from 0.64 to 0.70 across China.

  11. Providing Post-Treatment Support in an Outpatient Alcohol and Other Drug Treatment Context: A Qualitative Study of Staff Opinion

    ERIC Educational Resources Information Center

    Pulford, Justin; Black, Stella; Wheeler, Amanda; Sheridan, Janie; Adams, Peter

    2010-01-01

    This paper examines the post-treatment support practices, attitudes and preferences of outpatient alcohol and other drug (AOD) treatment staff as well as perceived barriers to implementing a post-treatment support service in an outpatient AOD treatment context. Data were collected via semi-structured interview and group discussion (n = 23).…

  12. An aryl-alcohol oxidase of Pleurotus sapidus: heterologous expression, characterization, and application in a 2-enzyme system.

    PubMed

    Galperin, Ilya; Javeed, Aysha; Luig, Hanno; Lochnit, Günter; Rühl, Martin

    2016-09-01

    Aryl-alcohol oxidases (AAOs) are enzymes supporting the degradation of lignin by fungal derived class II peroxidases produced by white-rot fungi. AAOs are able to generate H2O2 as a by-product via oxidation of an aryl-alcohol into its correspondent aldehyde. In this study, an AAO was heterologously expressed in a basidiomycete host for the first time. The gene for an AAO of the white-rot fungus Pleurotus sapidus, a close relative to the oyster mushroom Pleurotus ostreatus, was cloned into an expression vector and put under control of the promotor of the glyceraldehyde-3-phosphate dehydrogenase gene 2 (gpdII) of the button mushroom Agaricus bisporus. The expression vector was transformed into the model basidiomycete Coprinopsis cinerea, and several positive transformants were obtained. The best producing transformants were grown in shake-flasks and in a stirred tank reactor reaching enzymatic activities of up to 125 U L(-1) using veratryl alcohol as a substrate. The purified AAO was biochemically characterized and compared to the previously described native and recombinant AAOs from other Pleurotus species. In addition, a two-enzyme system comprising a dye-decolorizing peroxidase (DyP) from Mycetinis scorodonius and the P. sapidus AAO was successfully employed to bleach the anthraquinone dye Reactive Blue 5.

  13. Evaluation of an Online Alcohol Education Program for First-Time-in-College Students

    ERIC Educational Resources Information Center

    Reiss, Elayne R.

    2010-01-01

    Alcohol use among college students has maintained its place as a major issue in American higher education since its inception. Although dangerous drinking has always proliferated among college students, institutions have only provided alcohol and other drug (AOD) education and interventions encouraging students to adapt less harmful habits for a…

  14. Dust storm events over Delhi: verification of dust AOD forecasts with satellite and surface observations

    NASA Astrophysics Data System (ADS)

    Singh, Aditi; Iyengar, Gopal R.; George, John P.

    2016-05-01

    Thar desert located in northwest part of India is considered as one of the major dust source. Dust storms originate in Thar desert during pre-monsoon season, affects large part of Indo-Gangetic plains. High dust loading causes the deterioration of the ambient air quality and degradation in visibility. Present study focuses on the identification of dust events and verification of the forecast of dust events over Delhi and western part of IG Plains, during the pre-monsoon season of 2015. Three dust events have been identified over Delhi during the study period. For all the selected days, Terra-MODIS AOD at 550 nm are found close to 1.0, while AURA-OMI AI shows high values. Dust AOD forecasts from NCMRWF Unified Model (NCUM) for the three selected dust events are verified against satellite (MODIS) and ground based observations (AERONET). Comparison of observed AODs at 550 nm from MODIS with NCUM predicted AODs reveals that NCUM is able to predict the spatial and temporal distribution of dust AOD, in these cases. Good correlation (~0.67) is obtained between the NCUM predicted dust AODs and location specific observations available from AERONET. Model under-predicted the AODs as compared to the AERONET observations. This may be mainly because the model account for only dust and no anthropogenic activities are considered. The results of the present study emphasize the requirement of more realistic representation of local dust emission in the model both of natural and anthropogenic origin, to improve the forecast of dust from NCUM during the dust events.

  15. Aerosol Optical Depth (AOD) Trends Over Bangladesh

    NASA Astrophysics Data System (ADS)

    Salam, A.

    2016-12-01

    An important omission in the Southeast Asian observing network is the border region with the Indian subcontinent. Significant amounts of pollution are generated and transported down the Indo-Gangenic Plain into the Bay of Bengal. High population density in a semi-arid region leads to the development of a complex mixture of absorbing pollution coupled with dust. Transport patterns of this mixture takes pollutants into Bangladesh, where more pollution is added to the atmosphere-leading to what is one of the highest non urban emission loading in the world (AOD500= 0.75 during the premonsoon season). Bangladesh is essentially a riverine country, and atmospheric outflow is over delta regions fed by over 500 rivers, including the Ganges, Bramaputra, Jamuna, and Padma systems forming the massive Meghna river. This combination of atmospheric and riverine components makes for an optically complex littoral region which challenges a host of environmental sensors and modeling systems. Data is needed to understand the sources, transport and optical characteristics of aerosol particles in the region. Dhaka (23.8103° N, 90.4125° E) is the capital of Bangladesh with a population of about 16 million. It has been growing rapidly with all the problem of a mega city. We have installed a sun photometer with NASA Aeronet project at the roof of the Chemistry Department, Dhaka University with other aerosol particles and gas measuring instruments. Bhola is an Island of the Bay of Bengal. It is surrounded by the Meghna River on the north and east, the Tatulia River on the west and the Bay of Bengal on the south. The observatory is located at Charfashion Bazar, Bhola (N 22o10´01″, E 90o45´00″, 3m asl). There is very little influence from traffic and industrial emissions. A Cimel sunphotometer (NASA AERONET) was installed for AOD measurements at this locations since 2013. Aerosol optical depth (AOD) trends between 2012 and 2016 at two different locations (Dhaka and Bhola) will be

  16. A computer-assisted motivational social network intervention to reduce alcohol, drug and HIV risk behaviors among Housing First residents.

    PubMed

    Kennedy, David P; Hunter, Sarah B; Chan Osilla, Karen; Maksabedian, Ervant; Golinelli, Daniela; Tucker, Joan S

    2016-03-15

    Individuals transitioning from homelessness to housing face challenges to reducing alcohol, drug and HIV risk behaviors. To aid in this transition, this study developed and will test a computer-assisted intervention that delivers personalized social network feedback by an intervention facilitator trained in motivational interviewing (MI). The intervention goal is to enhance motivation to reduce high risk alcohol and other drug (AOD) use and reduce HIV risk behaviors. In this Stage 1b pilot trial, 60 individuals that are transitioning from homelessness to housing will be randomly assigned to the intervention or control condition. The intervention condition consists of four biweekly social network sessions conducted using MI. AOD use and HIV risk behaviors will be monitored prior to and immediately following the intervention and compared to control participants' behaviors to explore whether the intervention was associated with any systematic changes in AOD use or HIV risk behaviors. Social network health interventions are an innovative approach for reducing future AOD use and HIV risk problems, but little is known about their feasibility, acceptability, and efficacy. The current study develops and pilot-tests a computer-assisted intervention that incorporates social network visualizations and MI techniques to reduce high risk AOD use and HIV behaviors among the formerly homeless. CLINICALTRIALS. NCT02140359.

  17. 5-hydroxymethylfurfural conversion by fungal aryl-alcohol oxidase and unspecific peroxygenase.

    PubMed

    Carro, Juan; Ferreira, Patricia; Rodríguez, Leonor; Prieto, Alicia; Serrano, Ana; Balcells, Beatriz; Ardá, Ana; Jiménez-Barbero, Jesús; Gutiérrez, Ana; Ullrich, René; Hofrichter, Martin; Martínez, Angel T

    2015-08-01

    Oxidative conversion of 5-hydroxymethylfurfural (HMF) is of biotechnological interest for the production of renewable (lignocellulose-based) platform chemicals, such as 2,5-furandicarboxylic acid (FDCA). To the best of our knowledge, the ability of fungal aryl-alcohol oxidase (AAO) to oxidize HMF is reported here for the first time, resulting in almost complete conversion into 2,5-formylfurancarboxylic acid (FFCA) in a few hours. The reaction starts with alcohol oxidation, yielding 2,5-diformylfuran (DFF), which is rapidly converted into FFCA by carbonyl oxidation, most probably without leaving the enzyme active site. This agrees with the similar catalytic efficiencies of the enzyme with respect to oxidization of HMF and DFF, and its very low activity on 2,5-hydroxymethylfurancarboxylic acid (which was not detected by GC-MS). However, AAO was found to be unable to directly oxidize the carbonyl group in FFCA, and only modest amounts of FDCA are formed from HMF (most probably by chemical oxidation of FFCA by the H2 O2 previously generated by AAO). As aldehyde oxidation by AAO proceeds via the corresponding geminal diols (aldehyde hydrates), the various carbonyl oxidation rates may be related to the low degree of hydration of FFCA compared with DFF. The conversion of HMF was completed by introducing a fungal unspecific heme peroxygenase that uses the H2 O2 generated by AAO to transform FFCA into FDCA, albeit more slowly than the previous AAO reactions. By adding this peroxygenase when FFCA production by AAO has been completed, transformation of HMF into FDCA may be achieved in a reaction cascade in which O2 is the only co-substrate required, and water is the only by-product formed. © 2014 The Authors. FEBS Journal published by John Wiley & Sons Ltd on behalf of FEBS.

  18. Relevance of Campus Climate for Alcohol and Other Drug Use among LGBTQ Community College Students: A Statewide Qualitative Assessment

    ERIC Educational Resources Information Center

    Manning, Patricia; Pring, Lauren; Glider, Peggy

    2012-01-01

    Literature suggests that individuals who identify as LGBTQ may engage in more alcohol and other drug (AOD) use/abuse than others. Little data is available about these populations on college campuses where AOD use may be seen as part of the general campus climate and culture. This article will describe a qualitative needs assessment conducted on 10…

  19. A novel method to improve MODIS AOD retrievals in cloudy pixels using an analog ensemble approach

    NASA Astrophysics Data System (ADS)

    Kumar, R.; Raman, A.; Delle Monache, L.; Alessandrini, S.; Cheng, W. Y. Y.; Gaubert, B.; Arellano, A. F.

    2016-12-01

    Particulate matter (PM) concentrations are one of the fundamental indicators of air quality. Earth orbiting satellite platforms acquire column aerosol abundance that can in turn provide information about the PM concentrations. One of the serious limitations of column aerosol retrievals from low earth orbiting satellites is that these algorithms are based on clear sky assumptions. They do not retrieve AOD in cloudy pixels. After filtering cloudy pixels, these algorithms also arbitrarily remove brightest and darkest 25% of remaining pixels over ocean and brightest and darkest 50% pixels over land to filter any residual contamination from clouds. This becomes a critical issue especially in regions that experience monsoon, like Asia and North America. In case of North America, monsoon season experiences wide variety of extreme air quality events such as fires in California and dust storms in Arizona. Assessment of these episodic events warrants frequent monitoring of aerosol observations from remote sensing retrievals. In this study, we demonstrate a method to fill in cloudy pixels in Moderate Imaging Resolution Spectroradiometer (MODIS) AOD retrievals based on ensembles generated using an analog-based approach (AnEn). It provides a probabilistic distribution of AOD in cloudy pixels using historical records of model simulations of meteorological predictors such as AOD, relative humidity, and wind speed, and past observational records of MODIS AOD at a given target site. We use simulations from a coupled community weather forecasting model with chemistry (WRF-Chem) run at a resolution comparable to MODIS AOD. Analogs selected from summer months (June, July) of 2011-2013 from model and corresponding observations are used as a training dataset. Then, missing AOD retrievals in cloudy pixels in the last 31 days of the selected period are estimated. Here, we use AERONET stations as target sites to facilitate comparison against in-situ measurements. We use two approaches to

  20. Assessment of Alcohol and Other Drug Use Behaviors in Health Professions Students

    ERIC Educational Resources Information Center

    Baldwin, Jeffrey N.; Scott, David M.; Agrawal, Sangeeta; Bartek, Jean K.; Davis-Hall, R. Ellen; Reardon, Thomas P.; DeSimone, Edward M., II

    2006-01-01

    Alcohol and other drug (AOD) use behaviors of health professions students (HPS) were assessed by surveying both university-based HPS and other nursing programs in a Midwestern state in 1999. Response was 2,646 (56.4%) of surveyed students. Family history of alcohol-related and drug-related problems were reported by 39.8% and 13.9%, respectively,…

  1. Problem solving styles among people who use alcohol and other drugs in South Africa.

    PubMed

    Sorsdahl, Katherine; Stein, Dan J; Carrara, Henri; Myers, Bronwyn

    2014-01-01

    The present study examines the relationship between problem-solving styles, socio-demographic variables and risk of alcohol and other drug (AOD)-related problems among a South African population. The Social Problem-Solving Inventory-Revised, Center for Epidemiologic Studies Depression Scale (CES-D) and the Alcohol, Smoking and Substance Involvement Screening Test (ASSIST) were administered to a convenience sample of 1000 respondents. According to the ASSIST, 32% and 49% of respondents met criteria for moderate to high risk of alcohol use and illicit drug use respectively. After adjusting for the effects of other variables in the model, respondents who were of "Coloured" ancestry (PR=1.20, 95% CI 1.0-1.4), male (PR=1.19, 95% CI 1.04-1.37), older (PR=1.01, 95% CI 1.00-1.02), who adopted an avoidance style of coping with problems (PR=1.03, 95% CI 1.01-1.05) and who met criteria for depression (PR=1.42, 95% CI 1.12-1.79) were more likely to be classified as having risky AOD use. This suggests that interventions to improve problem solving and provide people with cognitive strategies to cope better with their problems may hold promise for reducing risky AOD use. © 2013.

  2. Assimilating AOD retrievals from GOCI and VIIRS to forecast surface PM2.5 episodes over Eastern China

    NASA Astrophysics Data System (ADS)

    Pang, Jiongming; Liu, Zhiquan; Wang, Xuemei; Bresch, Jamie; Ban, Junmei; Chen, Dan; Kim, Jhoon

    2018-04-01

    In this study, Geostationary Ocean Color Imager (GOCI) AOD and Visible Infrared Imaging Radiometer Suite (VIIRS) AOD data were assimilated to forecast surface PM2.5 concentrations over Eastern China, by using the three-dimensional variational (3DAVR) data assimilation (DA) system, to compare DA impacts by assimilating AOD retrievals from these two types of satellites. Three experiments were conducted, including a CONTROL without the AOD assimilation, and GOCIDA and VIIRSDA with the assimilation of AOD retrievals from GOCI and VIIRS, respectively. By utilizing the Weather Research and Forecasting with Chemistry (WRF/Chem) model, 48-h forecasts were initialized at each 06 UTC from 19 November to 06 December 2013. These forecasts were evaluated with 248 ground-based measurements from the air quality monitoring network across 67 China cities. The results show that overall the CONTROL underestimated surface PM2.5 concentrations, especially over Jing-Jin-Ji (JJJ) region and Yangtze River Delta (YRD) region. Both the GOCIDA and VIIRSDA produced higher surface PM2.5 concentrations mainly over Eastern China, which fits well with the PM2.5 measurements at these eastern sites, with more than 8% error reductions (ER). Moreover, compared to CONTROL, GOCIDA reduced 14.0% and 6.4% error on JJJ region and YRD region, respectively, while VIIRSDA reduced respectively 2.0% and 13.4% error over the corresponding areas. During the heavy polluted period, VIIRSDA improved all sites within YRD region, and GOCIDA enhanced 84% sites. Meanwhile, GOCIDA improved 84% sites on JJJ region, while VIIRSDA did not affect that region. These geographic distinctions might result from spatial dissimilarity between GOCI AOD and VIIRS AOD at time intervals. Moreover, the larger increment produced by AOD DA under stable meteorological conditions could lead to a longer duration (e.g., 1-2 days, > 2 days) of AOD DA impacts. Even though with AOD DA, surface PM2.5 concentrations were still underestimated

  3. Following the money: Mapping the sources and funding flows of alcohol and other drug treatment in Australia.

    PubMed

    Chalmers, Jenny; Ritter, Alison; Berends, Lynda; Lancaster, Kari

    2016-05-01

    The structures of health systems impact on patient outcomes. We present and analyse the first detailed mapping of who funds alcohol and other drug (AOD) treatment and the channels and intermediaries through which funding flows from the funding sources to treatment providers. The study involved a literature review of AOD treatment financing and existing diagrammatic representations of the structure of the Australian health system. We interviewed 190 key informants to particularise the AOD treatment sector, and undertook two case examples of government funded non-government organisations providing AOD treatment. Funding sources include the Australian and state and territory governments, philanthropy, fund-raising and clients themselves. While funding sources align with the health sector generally and the broader social services sector, the complexity of flows from source to treatment service and the number of intermediaries are noteworthy. So too are the many sources of funding drawn on by some treatment providers. Diversification is both beneficial and disadvantageous for non-government treatment providers, adding to administrative workloads, but smoothing the risk of funding shortfalls. Government funders benefit from sharing risk. Circuitous funding flows multiply the funding sources drawn on by services and put distance between the funding source and the service provider. This leads to concerns over lack of transparency about what is being purchased and challenges for the multiply funded service provider in maintaining programs and service models amid multiple and sometimes competing funding and accountability frameworks. [Chalmers J, Ritter A, Berends L, Lancaster K. Following the money: Mapping the sources and funding flows of alcohol and other drug treatment in Australia. Drug Alcohol Rev 2016;35:255-262]. © 2015 Australasian Professional Society on Alcohol and other Drugs.

  4. Heterogeneous Costs of Alcohol and Drug Problems Across Cities and Counties in California

    PubMed Central

    Miller, Ted R.; Nygaard, Peter; Gaidus, Andrew; Grube, Joel W.; Ponicki, William R.; Lawrence, Bruce A.; Gruenewald, Paul J.

    2017-01-01

    Background Estimates of economic and social costs related to alcohol and other drug (AOD) use and abuse are usually made at state and national levels. Ecological analyses demonstrate, however, that substantial variations exist in the incidence and prevalence of AOD use and problems including impaired driving, violence, and chronic disease between smaller geopolitical units like counties and cities. This study examines the ranges of these costs across counties and cities in California. Methods We used estimates of the incidence and prevalence of AOD use, abuse and related problems to calculate costs in 2010 dollars for all 58 counties and an ecological sample of 50 cities with populations between 50,000 and 500,000 persons in California. The estimates were built from archival and public-use survey data collected at state, county and city-levels over the years from 2009 to 2010. Results Costs related to alcohol use and related problems exceeded those related to illegal drugs across all counties and most cities in the study. Substantial heterogeneities in costs were observed between cities within counties. Conclusions AOD costs are heterogeneously distributed across counties and cities, reflecting the degree to which different populations are engaged in use and abuse across the state. These findings provide a strong argument for the distribution of treatment and prevention resources proportional to need. PMID:28208210

  5. A Xylenol Orange-Based Screening Assay for the Substrate Specificity of Flavin-Dependent para-Phenol Oxidases.

    PubMed

    Ewing, Tom A; van Noord, Aster; Paul, Caroline E; van Berkel, Willem J H

    2018-01-14

    Vanillyl alcohol oxidase (VAO) and eugenol oxidase (EUGO) are flavin-dependent enzymes that catalyse the oxidation of para -substituted phenols. This makes them potentially interesting biocatalysts for the conversion of lignin-derived aromatic monomers to value-added compounds. To facilitate their biocatalytic exploitation, it is important to develop methods by which variants of the enzymes can be rapidly screened for increased activity towards substrates of interest. Here, we present the development of a screening assay for the substrate specificity of para -phenol oxidases based on the detection of hydrogen peroxide using the ferric-xylenol orange complex method. The assay was used to screen the activity of VAO and EUGO towards a set of twenty-four potential substrates. This led to the identification of 4-cyclopentylphenol as a new substrate of VAO and EUGO and 4-cyclohexylphenol as a new substrate of VAO. Screening of a small library of VAO and EUGO active-site variants for alterations in their substrate specificity led to the identification of a VAO variant (T457Q) with increased activity towards vanillyl alcohol (4-hydroxy-3-methoxybenzyl alcohol) and a EUGO variant (V436I) with increased activity towards chavicol (4-allylphenol) and 4-cyclopentylphenol. This assay provides a quick and efficient method to screen the substrate specificity of para -phenol oxidases, facilitating the enzyme engineering of known para- phenol oxidases and the evaluation of the substrate specificity of novel para -phenol oxidases.

  6. Technical note: Intercomparison of three AATSR Level 2 (L2) AOD products over China

    NASA Astrophysics Data System (ADS)

    Che, Yahui; Xue, Yong; Mei, Linlu; Guang, Jie; She, Lu; Guo, Jianping; Hu, Yincui; Xu, Hui; He, Xingwei; Di, Aojie; Fan, Cheng

    2016-08-01

    One of four main focus areas of the PEEX initiative is to establish and sustain long-term, continuous, and comprehensive ground-based, airborne, and seaborne observation infrastructure together with satellite data. The Advanced Along-Track Scanning Radiometer (AATSR) aboard ENVISAT is used to observe the Earth in dual view. The AATSR data can be used to retrieve aerosol optical depth (AOD) over both land and ocean, which is an important parameter in the characterization of aerosol properties. In recent years, aerosol retrieval algorithms have been developed both over land and ocean, taking advantage of the features of dual view, which can help eliminate the contribution of Earth's surface to top-of-atmosphere (TOA) reflectance. The Aerosol_cci project, as a part of the Climate Change Initiative (CCI), provides users with three AOD retrieval algorithms for AATSR data, including the Swansea algorithm (SU), the ATSR-2ATSR dual-view aerosol retrieval algorithm (ADV), and the Oxford-RAL Retrieval of Aerosol and Cloud algorithm (ORAC). The validation team of the Aerosol-CCI project has validated AOD (both Level 2 and Level 3 products) and AE (Ångström Exponent) (Level 2 product only) against the AERONET data in a round-robin evaluation using the validation tool of the AeroCOM (Aerosol Comparison between Observations and Models) project. For the purpose of evaluating different performances of these three algorithms in calculating AODs over mainland China, we introduce ground-based data from CARSNET (China Aerosol Remote Sensing Network), which was designed for aerosol observations in China. Because China is vast in territory and has great differences in terms of land surfaces, the combination of the AERONET and CARSNET data can validate the L2 AOD products more comprehensively. The validation results show different performances of these products in 2007, 2008, and 2010. The SU algorithm performs very well over sites with different surface conditions in mainland China

  7. Estimation of surface-level PM concentration based on aerosol type classification and near-surface AOD over Korea

    NASA Astrophysics Data System (ADS)

    Kim, Kwanchul; Noh, Youngmin; Lee, Kwon H.

    2016-04-01

    Surface-level PM distribution was estimated from the satellite aerosol optical depth (AOD) products, taking the account of aerosol type classification and near-surface AOD over Jeju, Korea. For this purpose, data from various instruments such as satellites, sunphotometer, and Micro-pulse Lidar (MPL) was used during March 2008 and October 2009. Initial analyses of comparison with sunphotometer AOD and PM concentration showed some relatively poor relationship over Jeju, Korea. Since the AERONET L2 data has significant number of observations with high AOT values paired to low surface-level PM values, which were believed to be the effect of long-rage transport aerosols like as Asian dust and biomass burning. Stronger correlations (exceeding R = 0.8) were obtained by screening long-rage transport aerosols and calculating near-surface AOT considering aerosol profiles data from MPL and HYSPLIT air mass trajectory. The relationship found between corrected satellite observed AOD and surface-level PM concentration over Jeju is very similar. An approach to reduce the discrepancy between satellite observed AOD and PM concentration is demonstrated by tuning thresholds used to detect aerosol type from sunphotometer inversion data. Finally, the satellite observed AOD-surface PM concentration correlation is significantly improved. Our study clearly demonstrates that satellite observed AOD is a good surrogate for monitoring PM air quality over Korea.

  8. Comparison of AOD, AAOD and column single scattering albedo from AERONET retrievals and in situ profiling measurements

    NASA Astrophysics Data System (ADS)

    Andrews, Elisabeth; Ogren, John A.; Kinne, Stefan; Samset, Bjorn

    2017-05-01

    Here we present new results comparing aerosol optical depth (AOD), aerosol absorption optical depth (AAOD) and column single scattering albedo (SSA) obtained from in situ vertical profile measurements with AERONET ground-based remote sensing from two rural, continental sites in the US. The profiles are closely matched in time (within ±3 h) and space (within 15 km) with the AERONET retrievals. We have used Level 1.5 inversion retrievals when there was a valid Level 2 almucantar retrieval in order to be able to compare AAOD and column SSA below AERONET's recommended loading constraint (AOD > 0.4 at 440 nm). While there is reasonable agreement for the AOD comparisons, the direct comparisons of in situ-derived to AERONET-retrieved AAOD (or SSA) reveal that AERONET retrievals yield higher aerosol absorption than obtained from the in situ profiles for the low aerosol optical depth conditions prevalent at the two study sites. However, it should be noted that the majority of SSA comparisons for AOD440 > 0.2 are, nonetheless, within the reported SSA uncertainty bounds. The observation that, relative to in situ measurements, AERONET inversions exhibit increased absorption potential at low AOD values is generally consistent with other published AERONET-in situ comparisons across a range of locations, atmospheric conditions and AOD values. This systematic difference in the comparisons suggests a bias in one or both of the methods, but we cannot assess whether the AERONET retrievals are biased towards high absorption or the in situ measurements are biased low. Based on the discrepancy between the AERONET and in situ values, we conclude that scaling modeled black carbon concentrations upwards to match AERONET retrievals of AAOD should be approached with caution as it may lead to aerosol absorption overestimates in regions of low AOD. Both AERONET retrievals and in situ measurements suggest there is a systematic relationship between SSA and aerosol amount (AOD or aerosol light

  9. New approach to the retrieval of AOD and its uncertainty from MISR observations over dark water

    NASA Astrophysics Data System (ADS)

    Witek, Marcin L.; Garay, Michael J.; Diner, David J.; Bull, Michael A.; Seidel, Felix C.

    2018-01-01

    A new method for retrieving aerosol optical depth (AOD) and its uncertainty from Multi-angle Imaging SpectroRadiometer (MISR) observations over dark water is outlined. MISR's aerosol retrieval algorithm calculates cost functions between observed and pre-simulated radiances for a range of AODs (from 0.0 to 3.0) and a prescribed set of aerosol mixtures. The previous version 22 (V22) operational algorithm considered only the AOD that minimized the cost function for each aerosol mixture and then used a combination of these values to compute the final, best estimate AOD and associated uncertainty. The new approach considers the entire range of cost functions associated with each aerosol mixture. The uncertainty of the reported AOD depends on a combination of (a) the absolute values of the cost functions for each aerosol mixture, (b) the widths of the cost function distributions as a function of AOD, and (c) the spread of the cost function distributions among the ensemble of mixtures. A key benefit of the new approach is that, unlike the V22 algorithm, it does not rely on empirical thresholds imposed on the cost function to determine the success or failure of a particular mixture. Furthermore, a new aerosol retrieval confidence index (ARCI) is established that can be used to screen high-AOD retrieval blunders caused by cloud contamination or other factors. Requiring ARCI ≥ 0.15 as a condition for retrieval success is supported through statistical analysis and outperforms the thresholds used in the V22 algorithm. The described changes to the MISR dark water algorithm will become operational in the new MISR aerosol product (V23), planned for release in 2017.

  10. New Approach to the Retrieval of AOD and its Uncertainty from MISR Observations Over Dark Water

    NASA Astrophysics Data System (ADS)

    Witek, M. L.; Garay, M. J.; Diner, D. J.; Bull, M. A.; Seidel, F.

    2017-12-01

    A new method for retrieving aerosol optical depth (AOD) and its uncertainty from Multi-angle Imaging SpectroRadiometer (MISR) observations over dark water is outlined. MISR's aerosol retrieval algorithm calculates cost functions between observed and pre-simulated radiances for a range of AODs (from 0.0 to 3.0) and a prescribed set of aerosol mixtures. The previous Version 22 (V22) operational algorithm considered only the AOD that minimized the cost function for each aerosol mixture, then used a combination of these values to compute the final, "best estimate" AOD and associated uncertainty. The new approach considers the entire range of cost functions associated with each aerosol mixture. The uncertainty of the reported AOD depends on a combination of a) the absolute values of the cost functions for each aerosol mixture, b) the widths of the cost function distributions as a function of AOD, and c) the spread of the cost function distributions among the ensemble of mixtures. A key benefit of the new approach is that, unlike the V22 algorithm, it does not rely on arbitrary thresholds imposed on the cost function to determine the success or failure of a particular mixture. Furthermore, a new Aerosol Retrieval Confidence Index (ARCI) is established that can be used to screen high-AOD retrieval blunders caused by cloud contamination or other factors. Requiring ARCI≥0.15 as a condition for retrieval success is supported through statistical analysis and outperforms the thresholds used in the V22 algorithm. The described changes to the MISR dark water algorithm will become operational in the new MISR aerosol product (V23), planned for release in 2017.

  11. Peroxisomal Targeting, Import, and Assembly of Alcohol Oxidase in Pichia pastoris

    PubMed Central

    Waterham, Hans R.; Russell, Kimberly A.; de Vries, Yne; Cregg, James M.

    1997-01-01

    Alcohol oxidase (AOX), the first enzyme in the yeast methanol utilization pathway is a homooctameric peroxisomal matrix protein. In peroxisome biogenesis-defective (pex) mutants of the yeast Pichia pastoris, AOX fails to assemble into active octamers and instead forms inactive cytoplasmic aggregates. The apparent inability of AOX to assemble in the cytoplasm contrasts with other peroxisomal proteins that are able to oligomerize before import. To further investigate the import of AOX, we first identified its peroxisomal targeting signal (PTS). We found that sequences essential for targeting AOX are primarily located within the four COOH-terminal amino acids of the protein leucine-alanine-arginine-phenylalanine COOH (LARF). To examine whether AOX can oligomerize before import, we coexpressed AOX without its PTS along with wild-type AOX and determined whether the mutant AOX could be coimported into peroxisomes. To identify the mutant form of AOX, the COOH-terminal LARF sequence of the protein was replaced with a hemagglutinin epitope tag (AOX–HA). Coexpression of AOX–HA with wild-type AOX (AOX-WT) did not result in an increase in the proportion of AOX–HA present in octameric active AOX, suggesting that newly synthesized AOX–HA cannot oligomerize with AOX-WT in the cytoplasm. Thus, AOX cannot initiate oligomerization in the cytoplasm, but must first be targeted to the organelle before assembly begins. PMID:9396748

  12. Discovery of a Xylooligosaccharide Oxidase from Myceliophthora thermophila C1.

    PubMed

    Ferrari, Alessandro R; Rozeboom, Henriëtte J; Dobruchowska, Justyna M; van Leeuwen, Sander S; Vugts, Aniek S C; Koetsier, Martijn J; Visser, Jaap; Fraaije, Marco W

    2016-11-04

    By inspection of the predicted proteome of the fungus Myceliophthora thermophila C1 for vanillyl-alcohol oxidase (VAO)-type flavoprotein oxidases, a putative oligosaccharide oxidase was identified. By homologous expression and subsequent purification, the respective protein could be obtained. The protein was found to contain a bicovalently bound FAD cofactor. By screening a large number of carbohydrates, several mono- and oligosaccharides could be identified as substrates. The enzyme exhibits a strong substrate preference toward xylooligosaccharides; hence it is named xylooligosaccharide oxidase (XylO). Chemical analyses of the product formed upon oxidation of xylobiose revealed that the oxidation occurs at C1, yielding xylobionate as product. By elucidation of several XylO crystal structures (in complex with a substrate mimic, xylose, and xylobiose), the residues that tune the unique substrate specificity and regioselectivity could be identified. The discovery of this novel oligosaccharide oxidase reveals that the VAO-type flavoprotein family harbors oxidases tuned for specific oligosaccharides. The unique substrate profile of XylO hints at a role in the degradation of xylan-derived oligosaccharides by the fungus M. thermophila C1. © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

  13. Evaluation of VIIRS AOD over North China Plain: biases from aerosol models

    NASA Astrophysics Data System (ADS)

    Zhu, J.; Xia, X.; Wang, J.; Chen, H.; Zhang, J.; Oo, M. M.; Holz, R.

    2014-12-01

    With the launch of the Visible Infrared Imaging Radiometer Suit (VIIRS) instrument onboard Suomi National Polar-orbiting Partnership(S-NPP) in late 2011, the aerosol products of VIIRS are receiving much attention.To date, mostevaluations of VIIRS aerosol productswere carried out about aerosol optical depth (AOD). To further assess the VIIRS AOD in China which is a heavy polluted region in the world,we made a comparison between VIIRS AOD and CE-318 radiometerobservation at the following three sites overNorth China Plain (NCP): metropolis-Beijing (AERONET), suburbs-XiangHe (AERONET) and regional background site- Xinglong (CARSNET).The results showed the VIIRS AOD at 550 nm has a positive mean bias error (MBE) of 0.14-0.15 and root mean square error (RMBE) 0.20. Among three sites, Beijing is mainly a source of bias with MBE 0.17-0.18 and RMBE 0.23-0.24, and this bias is larger than some recent global statics recently published in the literature. Further analysis shows that this large bias in VIIRS AOD overNCP may be partly caused by the aerosol model selection in VIIRS aerosol inversion. According to the retrieval of sky radiance from CE-318 at three sites, aerosols in NCP have high mean real part of refractive indices (1.52-1.53), large volume mean radius (0.17-0.18) and low concentration (0.04-0.09) of fine aerosol, and small mean radius (2.86-2.92) and high concentration (0.06-0.16) of coarse mode aerosol. These observation-based aerosol single scattering properties and size of fine and coarse aerosols differ fromthe aerosol properties used in VIIRSoperational algorithm.The dominant aerosol models used in VIIRS algorithm for these three sites are less polluted urban aerosol in Beijing and low-absorption smoke in other two sites, all of which don't agree with the high imaginary part of refractive indices from CE-318 retrieval. Therefore, the aerosol models in VIIRS algorithm are likely to be refined in NCP region.

  14. Relationship between AOD and synoptic circulation over the Eastern Mediterranean: A comparison between subjective and objective classifications

    NASA Astrophysics Data System (ADS)

    Bodenheimer, Shalev; Nirel, Ronit; Lensky, Itamar M.; Dayan, Uri

    2018-03-01

    The Eastern Mediterranean (EM) Basin is strongly affected by dust originating from two of the largest world sources: The Sahara Desert and the Arabian Peninsula. Climatologically, the distribution pattern of aerosol optical depth (AOD), as proxy to particulate matter (PM), is known to be correlated with synoptic circulation. The climatological relationship between circulation type classifications (CTCs) and AOD levels over the EM Basin ("synoptic skill") was examined for the years 2000-2014. We compared the association between subjective (expert-based) and objective (fully automated) classifications and AOD using autoregressive models. After seasonal adjustment, the mean values of R2 for the different methods were similar. However, the distinct spatial pattern of the R2 values suggests that subjective classifications perform better in their area of expertise, specifically in the southeast region of the study area, while, objective CTCs had better synoptic skill over the northern part of the EM. This higher synoptic skill of subjective CTCs stem from their ability to identify distinct circulation types (e.g. Sharav lows and winter lows) that are infrequent but are highly correlated with AOD. Notably, a simple CTC based on seasonality rather than meteorological parameters predicted well AOD levels, especially over the south-eastern part of the domain. Synoptic classifications that are area-oriented are likely better predictors of AOD and possibly other environmental variables.

  15. Prevalence and pathways of recovery from drug and alcohol problems in the United States population: Implications for practice, research, and policy.

    PubMed

    Kelly, John F; Bergman, Brandon; Hoeppner, Bettina B; Vilsaint, Corrie; White, William L

    2017-12-01

    Alcohol and other drug (AOD) problems confer a global, prodigious burden of disease, disability, and premature mortality. Even so, little is known regarding how, and by what means, individuals successfully resolve AOD problems. Greater knowledge would inform policy and guide service provision. Probability-based survey of US adult population estimating: 1) AOD problem resolution prevalence; 2) lifetime use of "assisted" (i.e., treatment/medication, recovery services/mutual help) vs. "unassisted" resolution pathways; 3) correlates of assisted pathway use. Participants (response=63.4% of 39,809) responding "yes" to, "Did you use to have a problem with alcohol or drugs but no longer do?" assessed on substance use, clinical histories, problem resolution. Weighted prevalence of problem resolution was 9.1%, with 46% self-identifying as "in recovery"; 53.9% reported "assisted" pathway use. Most utilized support was mutual-help (45.1%,SE=1.6), followed by treatment (27.6%,SE=1.4), and emerging recovery support services (21.8%,SE=1.4), including recovery community centers (6.2%,SE=0.9). Strongest correlates of "assisted" pathway use were lifetime AOD diagnosis (AOR=10.8[7.42-15.74], model R2=0.13), drug court involvement (AOR=8.1[5.2-12.6], model R2=0.10), and, inversely, absence of lifetime psychiatric diagnosis (AOR=0.3[0.2-0.3], model R2=0.10). Compared to those with primary alcohol problems, those with primary cannabis problems were less likely (AOR=0.7[0.5-0.9]) and those with opioid problems were more likely (AOR=2.2[1.4-3.4]) to use assisted pathways. Indices related to severity were related to assisted pathways (R2<0.03). Tens of millions of Americans have successfully resolved an AOD problem using a variety of traditional and non-traditional means. Findings suggest a need for a broadening of the menu of self-change and community-based options that can facilitate and support long-term AOD problem resolution. Copyright © 2017 Elsevier B.V. All rights reserved.

  16. Daily Estimation of High Resolution PM2.5 Concentrations over BTH area by Fusing MODIS AOD and Ground Observations

    NASA Astrophysics Data System (ADS)

    Lyu, Baolei; Hu, Yongtao; Chang, Howard; Russell, Armistead; Bai, Yuqi

    2017-04-01

    The satellite-borne Moderate Resolution Imaging Spectroradiometer (MODIS) aerosol optical depth (AOD) is often used to predict ground-level fine particulate matter (PM2.5) concentrations. The associated estimation accuracy is always reduced by AOD missing values and by insufficiently accounting for the spatio-temporal PM2.5 variations. This study aims to estimate PM2.5 concentrations at a high resolution with enhanced accuracy by fusing MODIS AOD and ground observations in the polluted and populated Beijing-Tianjin-Hebei (BTH) area of China in 2014 and 2015. A Bayesian-based statistical downscaler was employed to model the spatio-temporally varied AOD-PM2.5 relationships. We resampled a 3 km MODIS AOD product to a 4 km resolution in a Lambert conic conformal projection, to assist comparison and fusion with CMAQ predictions. A two-step method was used to fill the missing AOD values to obtain a full AOD dataset with complete spatial coverage. The downscaler has a relatively good performance in the fitting procedure (R2 = 0.75) and in the cross validation procedure (with two evaluation methods, R2 = 0.58 by random method and R2 = 0.47 by city-specific method). The number of missing AOD values was serious and related to elevated PM2.5 concentrations. The gap-filled AOD values corresponded well with our understanding of PM2.5 pollution conditions in BTH. The prediction accuracy of PM2.5 concentrations were improved in terms of their annual and seasonal mean. As a result of its fine spatio-temporal resolution and complete spatial coverage, the daily PM2.5 estimation dataset could provide extensive and insightful benefits to related studies in the BTH area. This may include understanding the formation processes of regional PM2.5 pollution episodes, evaluating daily human exposure, and establishing pollution controlling measures.

  17. Estimation of PM2.5 and PM10 using ground-based AOD measurements during KORUS-AQ campaign

    NASA Astrophysics Data System (ADS)

    Koo, J. H.; Kim, J.; Kim, S.; Go, S.; Lee, S.; Lee, H.; Mok, J.; Hong, J.; Lee, J.; Eck, T. F.; Holben, B. N.

    2017-12-01

    During the KORUS-AQ campaign (2 May - 12 June, 2016), aerosol optical depth (AOD) was obtained at multiple channels using various ground-based instruments at Yonsei University, Seoul: AERONET sunphotometer, SKYNET skyradiometer, Brewer spectrophotometer, and multi-filter rotating shadowband radiometer (MFRSR). At the same location, planetary boundary layer (PBL) height and vertical profile of backscattering coefficients also can be obtained based on the celiometer measurements. Using celiometer products and various AODs, we try to estimate the amount of particular matter (PM2.5 and PM10) and validate with in-situ surface PM2.5 and PM10 measurements from AIRKOREA network. Direct comparison between PM2.5 and AOD reveals that the ultraviolet(UV) channel AOD has better correlations, due to the higher sensitivity of short wavelength to the fine-mode particle. In contrast, PM10 shows the highest correlation with the near-infrared(NIR) AOD. Next, we extract the boundary-layer portion of AOD using either PBL height or vertical profile of backscattering coefficients to compare with PM2.5 and PM10. Both results enhance the correlation, but consideration of weighting factor calculated from backscattering coefficients shows larger contribution to the correlation increase. Finally, we performed the multiple linear regression to estimate PM2.5 and PM10 using AODs. Consideration of meteorology (temperature, wind speed, and relative humidity) can enhance the correlation and also O3 and NO2 consideration highly contributes to the high correlation. This finding implies the importance to consider the ambient condition of secondary aerosol formation related to the PM2.5 variation. Multiple regression model finally finds the correlation 0.7-0.8, and diminishes the wavelength-dependent correlation patterns.

  18. Continuous aryl alcohol oxidase production under growth-limited conditions using a trickle bed reactor.

    PubMed

    Pardo-Planas, Oscar; Atiyeh, Hasan K; Prade, Rolf A; Müller, Michael; Wilkins, Mark R

    2018-05-01

    An A. nidulans strain with a pyridoxine marker was used for continuous production of aryl alcohol oxidase (AAO) in a trickle bed reactor (TBR). Modified medium with reduced zinc, no copper, and 5 g/L ascorbic acid that reduced melanin production and increased AAO productivity under growth limited conditions was used. Two air flow rates, 0.11 L/min (0.1 vvm) and 1.1 L/min (1.0 vvm) were tested. More melanin formation and reduced protein productivity were observed with air flow rate of 1.1 L/min. Three random packings were used as support for the fungus inside the TBR column, two of which were hydrophobic and one which was hydrophilic, and three different dilution rates were tested. The use of GEA BCN 030 hydrophobic packing resulted in greater AAO yield and productivity than the other packings. Increasing dilution rates favored melanin formation and citric, lactic and succinic acid accumulation, which decreased AAO yield and productivity. Copyright © 2018 Elsevier Ltd. All rights reserved.

  19. "I've Had a Pretty Tough Life but That's Not Why I Do This": Narratives of Autonomy and Control among Alcohol and Drug Service-Engaged Early Teenagers

    ERIC Educational Resources Information Center

    MacLean, Sarah J.; Bruun, Andrew; Mallett, Shelley

    2013-01-01

    The provision of alcohol and other drug (AOD) programmes in Australia targeting a broad age range of young people may inadvertently obscure the particular service needs of early teenagers. In this study, we describe four main accounts of substance use identified through interviews with 20 AOD service-engaged participants in Victoria, aged from 13…

  20. Innovative Recruitment Using Online Networks: Lessons Learned From an Online Study of Alcohol and Other Drug Use Utilizing a Web-Based, Respondent-Driven Sampling (webRDS) Strategy

    PubMed Central

    Bauermeister, José A.; Zimmerman, Marc A.; Johns, Michelle M.; Glowacki, Pietreck; Stoddard, Sarah; Volz, Erik

    2012-01-01

    Objective: We used a web version of Respondent-Driven Sampling (webRDS) to recruit a sample of young adults (ages 18–24) and examined whether this strategy would result in alcohol and other drug (AOD) prevalence estimates comparable to national estimates (National Survey on Drug Use and Health [NSDUH]). Method: We recruited 22 initial participants (seeds) via Facebook to complete a web survey examining AOD risk correlates. Sequential, incentivized recruitment continued until our desired sample size was achieved. After correcting for webRDS clustering effects, we contrasted our AOD prevalence estimates (past 30 days) to NSDUH estimates by comparing the 95% confidence intervals of prevalence estimates. Results: We found comparable AOD prevalence estimates between our sample and NSDUH for the past 30 days for alcohol, marijuana, cocaine, Ecstasy (3,4-methylenedioxymethamphetamine, or MDMA), and hallucinogens. Cigarette use was lower than NSDUH estimates. Conclusions: WebRDS may be a suitable strategy to recruit young adults online. We discuss the unique strengths and challenges that may be encountered by public health researchers using webRDS methods. PMID:22846248

  1. Innovative recruitment using online networks: lessons learned from an online study of alcohol and other drug use utilizing a web-based, respondent-driven sampling (webRDS) strategy.

    PubMed

    Bauermeister, José A; Zimmerman, Marc A; Johns, Michelle M; Glowacki, Pietreck; Stoddard, Sarah; Volz, Erik

    2012-09-01

    We used a web version of Respondent-Driven Sampling (webRDS) to recruit a sample of young adults (ages 18-24) and examined whether this strategy would result in alcohol and other drug (AOD) prevalence estimates comparable to national estimates (National Survey on Drug Use and Health [NSDUH]). We recruited 22 initial participants (seeds) via Facebook to complete a web survey examining AOD risk correlates. Sequential, incentivized recruitment continued until our desired sample size was achieved. After correcting for webRDS clustering effects, we contrasted our AOD prevalence estimates (past 30 days) to NSDUH estimates by comparing the 95% confidence intervals of prevalence estimates. We found comparable AOD prevalence estimates between our sample and NSDUH for the past 30 days for alcohol, marijuana, cocaine, Ecstasy (3,4-methylenedioxymethamphetamine, or MDMA), and hallucinogens. Cigarette use was lower than NSDUH estimates. WebRDS may be a suitable strategy to recruit young adults online. We discuss the unique strengths and challenges that may be encountered by public health researchers using webRDS methods.

  2. Can MODIS AOD be employed to derive PM2.5 in Beijing-Tianjin-Hebei over China?

    NASA Astrophysics Data System (ADS)

    Ma, Xiaoyan

    2017-04-01

    The fine particular matter (PM) concentrations in China have increased considerably due to the rapid economic growth and urbanization in the last few decades, especially in the most populated and industrialized regions. The Beijing-Tianjin-Hebei is one of the most polluted regions in China, so to monitor the PM2.5 concentrations over this region is quite critical for human health. By making use the new released hourly PM2.5 mass concentration from ground-based observations in Beijing-Tianjin-Hebei over China, and the collocated MODIS level 2 AOD data from April 2014 to March 2015, we explored the relation between surface PM2.5 mass concentration and MODIS AOD and possibility to derive the surface PM2.5 from satellite retrieval in the region. Our study show that the relation strongly depend on the seasons due to distinct seasonal characteristics of PM2.5 and AOD, with a relatively better correlation in spring and summertime than autumn and wintertime. Our analysis give an evidence that worse relationship and/or smaller number of sample in wintertime is associated with the significantly high PM2.5 concentration and a lot of missing data occurring in MODIS AOD, implying that current MODIS AOD retrieval scheme does not work very well in highly polluted cases. The derived PM2.5 mass concentration from MODIS AOD in summertime can basically capture the major observed features of the time series and about 20% large bias of the derived values compared to the observation is expected to be reduced once longer time period data is available and used for analysis.

  3. Characteristics of Gloeophyllum trabeum Alcohol Oxidase, an Extracellular Source of H2O2 in Brown Rot Decay of Wood▿

    PubMed Central

    Daniel, Geoffrey; Volc, Jindřich; Filonova, Lada; Plíhal, Ondřej; Kubátová, Elena; Halada, Petr

    2007-01-01

    A novel alcohol oxidase (AOX) has been purified from mycelial pellets of the wood-degrading basidiomycete Gloeophyllum trabeum and characterized as a homooctameric nonglycosylated protein with native and subunit molecular masses of 628 and 72.4 kDa, containing noncovalently bonded flavin adenine dinucleotide. The isolated AOX cDNA contained an open reading frame of 1,953 bp translating into a polypeptide of 651 amino acids displaying 51 to 53% identity with other published fungal AOX amino acid sequences. The enzyme catalyzed the oxidation of short-chain primary aliphatic alcohols with a preference for methanol (Km = 2.3 mM, kcat = 15.6 s−1). Using polyclonal antibodies and immunofluorescence staining, AOX was localized on liquid culture hyphae and extracellular slime in sections from degraded wood and on cotton fibers. Transmission electron microscopy immunogold labeling localized the enzyme in the hyphal periplasmic space and wall and on extracellular tripartite membranes and slime, while there was no labeling of hyphal peroxisomes. AOX was further shown to be associated with membranous or slime structures secreted by hyphae in wood fiber lumina and within the secondary cell walls of degraded wood fibers. The differences in AOX targeting compared to the known yeast peroxisomal localization were traced to a unique C-terminal sequence of the G. trabeum oxidase, which is apparently responsible for the protein's different translocation. The extracellular distribution and the enzyme's abundance and preference for methanol, potentially available from the demethylation of lignin, all point to a possible role for AOX as a major source of H2O2, a component of Fenton's reagent implicated in the generally accepted mechanisms for brown rot through the production of highly destructive hydroxyl radicals. PMID:17660304

  4. Estimation of the Cloud condensation nuclei concentration(CCN) and aerosol optical depth(AOD) relation in the Arctic region

    NASA Astrophysics Data System (ADS)

    Jung, C. H.; Yoon, Y. J.; Ahn, S. H.; Kang, H. J.; Gim, Y. T.; Lee, B. Y.

    2017-12-01

    Information of the spatial and temporal variations of cloud condensation nuclei (CCN) concentrations is important in estimating aerosol indirect effects. Generally, CCN aerosol is difficult to estimate using remote sensing methods. Although there are many CCN measurements data, extensive measurements of CCN are not feasible because of the complex nature of the operation and high cost, especially in the Arctic region. Thus, there have been many attempts to estimate CCN concentrations from more easily obtainable parameters such as aerosol optical depth (AOD) because AOD has the advantage of being readily observed by remote sensing from space by several sensors. For example, some form of correlation was derived between AOD and the number concentration of cloud condensation nuclei (CCN) through the comparison results from AERONET network and CCN measurements (Andreae 2009). In this study, a parameterization of CCN concentration as a function of AOD at 500 nm is given in the Arctic region. CCN data was collected during the period 2007-2013 at the Zeppelin observatory (78.91° N, 11.89° E, 474 masl). The AERONET network and MODIS AOD data are compared with ground measured CCN measurement and the relations between AOD and CCN are parameterized. The seasonal characteristics as well as long term trends are also considered. Through the measurement, CCN concentration remains high during spring because of aerosol transportation from the mid-latitudes, known as Arctic Haze. Lowest CCN number densities were observed during Arctic autumn and early winter when aerosol long-range transport into the Arctic is not effective and new particle formation ceases. The results show that the relation between AOD and CCN shows a different parameter depending on the seasonal aerosol and CCN characteristics. This seasonal different CCN-AOD relation can be interpreted as many physico-chemical aerosol properties including aerosol size distribution, composition. ReferenceAndreae, M. O. (2009

  5. Developing performance measures for alcohol and other drug services in managed care plans. Washington Circle Group.

    PubMed

    McCorry, F; Garnick, D W; Bartlett, J; Cotter, F; Chalk, M

    2000-11-01

    Monitoring the quality and availability of alcohol and other drug (AOD) services must be a central tenet of any health-related performance measurement system. The Washington Circle Group (WCG), which was convened by the Center for Substance Abuse Treatment Office of Managed Care in March 1998, has developed a core set of performance measures for AOD services for public- and private-sector health plans. It is also collaborating with a broad range of stakeholders to ensure widespread adoption of these performance measures by health plans, private employers, public payers, and accrediting organizations. Four domains were identified, with specific measures developed for each domain: (1) prevention/education, (2) recognition, (3) treatment (including initiation of alcohol and other plan services, linkage of detoxification and AOD plan services, treatment engagement, and interventions for family members/significant others), and (4) maintenance of treatment effects. Four measures that are based on administrative information from health plans and two measures that require a consumer survey of behavioral health care are undergoing extensive pilot testing. The WCG has reached out to a broad range of stakeholders in performance measurement and managed care to acquaint them with the measures and to promote their investigation and adoption. As results of pilot testing become available, these outreach efforts will continue. Performance measures for AOD services need to become an integral part of a comprehensive set of behavioral and physical health performance measures for managed care plans.

  6. Can MODIS AOD be employed to derive PM2.5 in Beijing-Tianjin-Hebei over China?

    NASA Astrophysics Data System (ADS)

    Ma, Xiaoyan; Wang, Jianying; Yu, Fangqun; Jia, Hailing; Hu, Yanan

    2016-11-01

    The fine particular matter (PM) concentrations in China have increased considerably due to the rapid economic growth and urbanization in the last few decades, especially in the most populated and industrialized regions. Beijing-Tianjin-Hebei is one of the most polluted regions in China, so to monitor the PM2.5 concentrations over this region is quite critical for human health. By making use the new released hourly PM2.5 mass concentration from ground-based observations in Beijing-Tianjin-Hebei over China, and collocated MODIS level 2 AOD data from April 2014 to March 2015, we explored the relation between surface PM2.5 mass concentration and MODIS AOD and possibility to derive the surface PM2.5 from satellite retrieval in the region. Our study show that the relation strongly depend on the seasons due to distinct seasonal characteristics of PM2.5 and AOD, with a relatively better correlation in spring and summertime (correlation coefficient r ranging from 0.52 to 0.79) than autumn and wintertime (r can be low as to 0.23 in site Baoding). Our analysis gave evidence that worse relationship and/or smaller number of sample in wintertime is associated with the significantly high PM2.5 concentration and a lot of missing data occurring in MODIS AOD, implying that current MODIS AOD retrieval scheme does not work very well in highly polluted cases. The derived PM2.5 mass concentration from MODIS AOD in summertime can basically capture the major observed features of the time series and about 20% large bias of the derived values compared to the observation is expected to be reduced if longer time period data is available and used for analysis.

  7. Mineralogical determination and geo-chemical modeling of chromium release from AOD slag: Distribution and leachability aspects.

    PubMed

    Li, Junguo; Liu, Bao; Zeng, Yanan; Wang, Ziming

    2017-01-01

    AOD (argon oxygen decarburization) slag, which is the by-product of the stainless steel refining process, is a recyclable slag because of its high content of calcium and silicon. The leaching toxicity cannot be ignored in the recycling process because the slag contains a certain amount of Cr. In this study, the mineral analysis, batch leaching tests and thermodynamic and kinetic modeling by PHREEQC combined with FactSage software were performed to explore the influence of the dissolution of primary minerals and the precipitation of secondary minerals on the elution of Cr from AOD slag. The results indicated that the main minerals in the original AOD slag are larnite, merwinite, pyroxene and periclase. Cr was dispersed in the mineral phases mentioned above. The simulation of Cr leaching controlled by Cr(III)-hydroxide corresponded better to the batch leaching tests, while the Cr leaching controlled by chromite or double control was underestimated. Increasing the L/S ratio enhances the pH of the leachate and restrains the elution of Cr from the AOD slag. Copyright © 2016 Elsevier Ltd. All rights reserved.

  8. Bienzyme biosensors for glucose, ethanol and putrescine built on oxidase and sweet potato peroxidase.

    PubMed

    Castillo, Jaime; Gáspár, Szilveszter; Sakharov, Ivan; Csöregi, Elisabeth

    2003-05-01

    Amperometric biosensors for glucose, ethanol, and biogenic amines (putrescine) were constructed using oxidase/peroxidase bienzyme systems. The H(2)O(2) produced by the oxidase in reaction with its substrate is converted into a measurable signal via a novel peroxidase purified from sweet potato peels. All developed biosensors are based on redox hydrogels formed of oxidases (glucose oxidase, alcohol oxidase, or amine oxidase) and the newly purified sweet potato peroxidase (SPP) cross-linked to a redox polymer. The developed electrodes were characterized (sensitivity, stability, and performances in organic medium) and compared with similarly built ones using the 'classical' horseradish peroxidase (HRP). The SPP-based electrodes displayed higher sensitivity and better detection limit for putrescine than those using HRP and were also shown to retain their activity in organic phase much better than the HPR based ones. The importance of attractive or repulsive electrostatic interactions between the peroxidases and oxidases (determined by their isoelectric points) were found to play an important role in the sensitivity of the obtained sensors.

  9. Oxidative Stress in Aortas of Patients with Advanced Occlusive and Aneurysmal Diseases.

    PubMed

    Lucas, Márcio L; Carraro, Cristina C; Belló-Klein, Adriane; Kalil, Antônio N; Aerts, Newton R; Carvalho, Fabiano B; Fernandes, Marilda C; Zettler, Claudio G

    2018-06-06

    Aortoiliac occlusive disease (AOD) and abdominal aortic aneurysm (AAA) are very important cardiovascular diseases that present different aspects of pathophysiology; however, oxidative stress and inflammatory response seem be relevant in both of them. Our objective was to evaluate oxidative damage and degree of inflammatory infiltrate in aortas of patients surgically treated for AOD and AAA. Levels of reactive oxygen species (ROS), nicotinamide adenine dinucleotide phosphate (NADPH) oxidase activity, and myeloperoxidase (MPO) expression as well as nitrite levels and superoxide dismutase (SOD) and catalase (CAT) activities were evaluated in aortas of patients with AOD (n = 16) or AAA (n = 14), while the control group was formed by cadaveric organ donors (n = 10). We also analyzed the degree of inflammatory infiltrate in these aortas. There was an increase in ROS levels and NADPH oxidase activity in patients with AOD and AAA when compared with the control group, and the AOD group demonstrated higher ROS production and NADPH oxidase activity and also nitrite levels when compared with the AAA group (P < 0.001). On the other hand, an increase of SOD activity in the AOD group and CAT activity in the AAA group was observed. Inflammatory infiltrate and MPO expression were higher in the AOD group when compared with the control group (P < 0.05). Oxidative stress is relevant in both AOD and AAA, though AOD presented higher ROS levels and NADPH activity. Increased activities of antioxidant enzymes may be a compensatory phenomenon which occurs in aortas of patients with AOD and AAA. Perhaps, a relationship between oxidative stress and degree of inflammatory infiltrate may exist in the pathophysiology of AOD and AAA. Copyright © 2018 Elsevier Inc. All rights reserved.

  10. Effect of quality chronic disease management for alcohol and drug dependence on addiction outcomes.

    PubMed

    Kim, Theresa W; Saitz, Richard; Cheng, Debbie M; Winter, Michael R; Witas, Julie; Samet, Jeffrey H

    2012-12-01

    We examined the effect of the quality of primary care-based chronic disease management (CDM) for alcohol and/or other drug (AOD) dependence on addiction outcomes. We assessed quality using (1) a visit frequency based measure and (2) a self-reported assessment measuring alignment with the chronic care model. The visit frequency based measure had no significant association with addiction outcomes. The self-reported measure of care-when care was at a CDM clinic-was associated with lower drug addiction severity. The self-reported assessment of care from any healthcare source (CDM clinic or elsewhere) was associated with lower alcohol addiction severity and abstinence. These findings suggest that high quality CDM for AOD dependence may improve addiction outcomes. Quality measures based upon alignment with the chronic care model may better capture features of effective CDM care than a visit frequency measure. Copyright © 2012 Elsevier Inc. All rights reserved.

  11. Bias Correction of MODIS AOD using DragonNET to obtain improved estimation of PM2.5

    NASA Astrophysics Data System (ADS)

    Gross, B.; Malakar, N. K.; Atia, A.; Moshary, F.; Ahmed, S. A.; Oo, M. M.

    2014-12-01

    MODIS AOD retreivals using the Dark Target algorithm is strongly affected by the underlying surface reflection properties. In particular, the operational algorithms make use of surface parameterizations trained on global datasets and therefore do not account properly for urban surface differences. This parameterization continues to show an underestimation of the surface reflection which results in a general over-biasing in AOD retrievals. Recent results using the Dragon-Network datasets as well as high resolution retrievals in the NYC area illustrate that this is even more significant at the newest C006 3 km retrievals. In the past, we used AERONET observation in the City College to obtain bias-corrected AOD, but the homogeneity assumptions using only one site for the region is clearly an issue. On the other hand, DragonNET observations provide ample opportunities to obtain better tuning the surface corrections while also providing better statistical validation. In this study we present a neural network method to obtain bias correction of the MODIS AOD using multiple factors including surface reflectivity at 2130nm, sun-view geometrical factors and land-class information. These corrected AOD's are then used together with additional WRF meteorological factors to improve estimates of PM2.5. Efforts to explore the portability to other urban areas will be discussed. In addition, annual surface ratio maps will be developed illustrating that among the land classes, the urban pixels constitute the largest deviations from the operational model.

  12. Fine Particulate Matter Predictions Using High Resolution Aerosol Optical Depth (AOD) Retrievals

    NASA Technical Reports Server (NTRS)

    Chudnovsky, Alexandra A.; Koutrakis, Petros; Kloog, Itai; Melly, Steven; Nordio, Francesco; Lyapustin, Alexei; Wang, Jujie; Schwartz, Joel

    2014-01-01

    To date, spatial-temporal patterns of particulate matter (PM) within urban areas have primarily been examined using models. On the other hand, satellites extend spatial coverage but their spatial resolution is too coarse. In order to address this issue, here we report on spatial variability in PM levels derived from high 1 km resolution AOD product of Multi-Angle Implementation of Atmospheric Correction (MAIAC) algorithm developed for MODIS satellite. We apply day-specific calibrations of AOD data to predict PM(sub 2.5) concentrations within the New England area of the United States. To improve the accuracy of our model, land use and meteorological variables were incorporated. We used inverse probability weighting (IPW) to account for nonrandom missingness of AOD and nested regions within days to capture spatial variation. With this approach we can control for the inherent day-to-day variability in the AOD-PM(sub 2.5) relationship, which depends on time-varying parameters such as particle optical properties, vertical and diurnal concentration profiles and ground surface reflectance among others. Out-of-sample "ten-fold" cross-validation was used to quantify the accuracy of model predictions. Our results show that the model-predicted PM(sub 2.5) mass concentrations are highly correlated with the actual observations, with out-of- sample R(sub 2) of 0.89. Furthermore, our study shows that the model captures the pollution levels along highways and many urban locations thereby extending our ability to investigate the spatial patterns of urban air quality, such as examining exposures in areas with high traffic. Our results also show high accuracy within the cities of Boston and New Haven thereby indicating that MAIAC data can be used to examine intra-urban exposure contrasts in PM(sub 2.5) levels.

  13. Links between alcohol and other drug problems and maltreatment among adolescent girls: perceived discrimination, ethnic identity, and ethnic orientation as moderators.

    PubMed

    Gray, Calonie M K; Montgomery, Marilyn J

    2012-05-01

    This study examined the links between maltreatment, posttraumatic stress symptoms, ethnicity-specific factors (i.e., perceived discrimination, ethnic identity, and ethnic orientation), and alcohol and/or other drug (AOD) problems among adolescent girls. These relations were examined using archived data from a community sample of 168 Black and Hispanic adolescent girls who participated in a school-based substance use intervention. The results revealed that maltreatment was linked to AOD problems, but only through its relation with posttraumatic stress symptoms; maltreatment was positively related to posttraumatic stress symptoms, which were positively related to AOD problems. Both perceived discrimination and ethnic orientation were significant moderators. Specifically, greater perceived discrimination was associated with an increased effect of maltreatment on posttraumatic stress symptoms. Ethnic orientation demonstrated protective properties in the relation between maltreatment and AOD problem severity, such that the effect of maltreatment on AOD problem severity was less for girls with average to high ethnic orientation compared to girls with low ethnic orientation. The findings of this study underscore the importance of developing interventions for Black and Hispanic girls that target maltreatment and AOD use concurrently and address ethnicity-specific factors. Copyright © 2012 Elsevier Ltd. All rights reserved.

  14. Links between alcohol and other drug problems and maltreatment among adolescent girls: Perceived discrimination, ethnic identity, and ethnic orientation as moderators

    PubMed Central

    Gray, Calonie M. K.; Montgomery, Marilyn J.

    2012-01-01

    Objectives This study examined the links between maltreatment, posttraumatic stress symptoms, ethnicity-specific factors (i.e., perceived discrimination, ethnic identity, and ethnic orientation), and alcohol and/or other drug (AOD) problems among adolescent girls. Methods These relations were examined using archived data from a community sample of 168 Black and Hispanic adolescent girls who participated in a school-based substance use intervention. Results The results revealed that maltreatment was linked to AOD problems, but only through its relation with posttraumatic stress symptoms; maltreatment was positively related to posttraumatic stress symptoms, which were positively related to AOD problems. Both perceived discrimination and ethnic orientation were significant moderators. Specifically, greater perceived discrimination was associated with an increased effect of maltreatment on posttraumatic stress symptoms. Ethnic orientation demonstrated protective properties in the relation between maltreatment and AOD problem severity, such that the effect of maltreatment on AOD problem severity was less for girls with average to high ethnic orientation compared to girls with low ethnic orientation. Conclusions The findings of this study underscore the importance of developing interventions for Black and Hispanic girls that target maltreatment and AOD use concurrently and address ethnicity-specific factors. PMID:22608406

  15. Assessment of function and clinical utility of alcohol and other drug web sites: An observational, qualitative study

    PubMed Central

    2011-01-01

    Background The increasing popularity and use of the internet makes it an attractive option for providing health information and treatment, including alcohol/other drug use. There is limited research examining how people identify and access information about alcohol or other drug (AOD) use online, or how they assess the usefulness of the information presented. This study examined the strategies that individuals used to identify and navigate a range of AOD websites, along with the attitudes concerning presentation and content. Methods Members of the general community in Brisbane and Roma (Queensland, Australia) were invited to participate in a 30-minute search of the internet for sites related to AOD use, followed by a focus group discussion. Fifty one subjects participated in the study across nine focus groups. Results Participants spent a maximum of 6.5 minutes on any one website, and less if the user was under 25 years of age. Time spent was as little as 2 minutes if the website was not the first accessed. Participants recommended that AOD-related websites should have an engaging home or index page, which quickly and accurately portrayed the site's objectives, and provided clear site navigation options. Website content should clearly match the title and description of the site that is used by internet search engines. Participants supported the development of a portal for AOD websites, suggesting that it would greatly facilitate access and navigation. Treatment programs delivered online were initially viewed with caution. This appeared to be due to limited understanding of what constituted online treatment, including its potential efficacy. Conclusions A range of recommendations arise from this study regarding the design and development of websites, particularly those related to AOD use. These include prudent use of text and information on any one webpage, the use of graphics and colours, and clear, uncluttered navigation options. Implications for future website

  16. 12-Step participation reduces medical use costs among adolescents with a history of alcohol and other drug treatment.

    PubMed

    Mundt, Marlon P; Parthasarathy, Sujaya; Chi, Felicia W; Sterling, Stacy; Campbell, Cynthia I

    2012-11-01

    Adolescents who attend 12-step groups following alcohol and other drug (AOD) treatment are more likely to remain abstinent and to avoid relapse post-treatment. We examined whether 12-step attendance is also associated with a corresponding reduction in health care use and costs. We used difference-in-difference analysis to compare changes in seven-year follow-up health care use and costs by changes in 12-step participation. Four Kaiser Permanente Northern California AOD treatment programs enrolled 403 adolescents, 13-18-years old, into a longitudinal cohort study upon AOD treatment entry. Participants self-reported 12-step meeting attendance at six-month, one-year, three-year, and five-year follow-up. Outcomes included counts of hospital inpatient days, emergency room (ER) visits, primary care visits, psychiatric visits, AOD treatment costs and total medical care costs. Each additional 12-step meeting attended was associated with an incremental medical cost reduction of 4.7% during seven-year follow-up. The medical cost offset was largely due to reductions in hospital inpatient days, psychiatric visits, and AOD treatment costs. We estimate total medical use cost savings at $145 per year (in 2010 U.S. dollars) per additional 12-step meeting attended. The findings suggest that 12-step participation conveys medical cost offsets for youth who undergo AOD treatment. Reduced costs may be related to improved AOD outcomes due to 12-step participation, improved general health due to changes in social network following 12-step participation, or better compliance to both AOD treatment and 12-step meetings. Copyright © 2012 Elsevier Ireland Ltd. All rights reserved.

  17. GRACE AOD1B Product Release 06: Long-Term Consistency and the Treatment of Atmospheric Tides

    NASA Astrophysics Data System (ADS)

    Dobslaw, Henryk; Bergmann-Wolf, Inga; Dill, Robert; Poropat, Lea; Flechtner, Frank

    2017-04-01

    The GRACE satellites orbiting the Earth at very low altitudes are affected by rapid changes in the Earth's gravity field caused by mass redistribution in atmosphere and oceans. To avoid temporal aliasing of such high-frequency variability into the final monthly-mean gravity fields, those effects are typically modelled during the numerical orbit integration by appling the 6-hourly GRACE Atmosphere and Ocean De-Aliasing Level-1B (AOD1B) a priori model. In preparation of the next GRACE gravity field re-processing currently performed by the GRACE Science Data System, a new version of AOD1B has been calculated. The data-set is based on 3-hourly surface pressure anomalies from ECMWF that have been mapped to a common reference orography by means of ECMWF's mean sea-level pressure diagnostic. Atmospheric tides as well as the corresponding oceanic response at the S1, S2, S3, and L2 frequencies and its annual modulations have been fitted and removed in order to retain the non-tidal variability only. The data-set is expanded into spherical harmonics complete up to degree and order 180. In this contribution, we will demonstrate that AOD1B RL06 is now free from spurious jumps in the time-series related to occasional changes in ECMWF's operational numerical weather prediction system. We will also highlight the rationale for separating tidal signals from the AOD1B coefficients, and will finally discuss the current quality of the AOD1B forecasts that have been introduced very recently for GRACE quicklook or near-realtime applications.

  18. Retrieval of AOD and PM2.5 Concentrations over Urban Areas of Shenyang City using MODIS Data

    NASA Astrophysics Data System (ADS)

    Wang, Z.

    2016-12-01

    Atmospheric aerosols play an important part in the Earth's radiation balance as well as global climate change, aerosols also have very important impact on environment as well as human and other organisms' health, PM2.5 and other small particle aerosols, can enter bronchi directly, thus causing bronchitis, cardiovascular disease, asthma and so on.Detection of AOD by satellite and remote sensing is currently one of the hotest issues , diffierent from the traditional monitoring method, this method has much more advantanges, for emample wide area coverage, fast and convenient etc. So it is possible for people to know the regional changes of AOD real time over large area. Now, detection aerosol by RS technology has reached a high level in marine and dense vegetation land areas, but result is not ideal for urban areas, the higher surface reflectance in urban areas is a bottleneck of AOD retrieval. Focus on the high surface reflectance and low accuracy of the AOD products of urban areas, this paper propose an algorithm coupled with surface reflectance to get red band surface reflectance, based on Dens Dark Vegetation algorithm and geometrical optics model theory, to distinguish urban reflectivity from other targets. Considering the appropriate aerosol model which adapt to season and other proper parameters, this paper uses 6S model to establish look-up table, thus retrieve AOD for urban as well as other high reflectance areas. This paper take Shenyang region as pilot area, then retrieve the AOD and PM2.5 concentration of Shenyang in 2015 based on MODIS data, thus get 1km resolution distribution map, and then analyzed the results in spatial, intensity and temporal. At last, real-time monitoring data from the ground monitor station is used to verify the outcome, the results have good accuracy and the the correlation reached 0.9004 when the weather is sunny. The research shows that this algorithm has relatively higher precision and certain universality. This method has

  19. Surface modification of polyvinyl alcohol/malonic acid nanofibers by gaseous dielectric barrier discharge plasma for glucose oxidase immobilization

    NASA Astrophysics Data System (ADS)

    Afshari, Esmail; Mazinani, Saeedeh; Ranaei-Siadat, Seyed-Omid; Ghomi, Hamid

    2016-11-01

    Polymeric nanofiber prepares a suitable situation for enzyme immobilization for variety of applications. In this research, we have fabricated polyvinyl alcohol (PVA)/malonic acid nanofibers using electrospinning. After fabrication of nanofibers, the effect of air, nitrogen, CO2, and argon DBD (dielectric barrier discharge) plasmas on PVA/malonic acid nanofibers were analysed. Among them, air plasma had the most significant effect on glucose oxidase (GOx) immobilization. Attenuated total reflectance-Fourier transform infrared (ATR-FTIR) spectrum analysis and X-ray photoelectron spectroscopy (XPS) results revealed that in case of air plasma modified nanofibers, the carboxyl groups on the surface are increased. The scanning electron microscopy (SEM) images showed that, after GOx immobilization, the modified nanofibers with plasma has retained its nanofiber structure. Finally, we analysed reusability and storage stability of GOx immobilized on plasma modified and unmodified nanofibers. The results were more satisfactory for modified nanofibers with respect to unmodified ones.

  20. Evaluation of monthwise and overall trends of AOD over Indian cities using MODIS Aqua and Terra retrievals

    NASA Astrophysics Data System (ADS)

    Banerjee, Subhasis; Ghosh, Sanjay

    2016-07-01

    Atmospheric aerosols have been shown to have profound impact on climate system and human health. Regular and systematic monitoring of ambient air is thus necessary in order to asses its impact. There are several ground based stations worldwide employed in this service but still their numbers are inadequate and it is even almost impossible to have such stations at difficult geographical terrains and take measurement throughout the year. Aerosol optical depth or AOD, which is a measure of extinction of incoming solar radiation, serves as proxy to atmospheric aerosol loading. Various sensors onboard different satellites take routine measurement of AOD throughout the year. Satellite based AOD is used in many studies due to their wide coverage and availability for a longer time period. Satellite measures reflected solar radiation at the top of the atmosphere. Column integrated value of aerosol are routinely estimated from those measurements using suitable inversion algorithms. MODIS instrument onboard Aqua and Terra satellites of Earth Observing System takes routine measurement in wide spectral range. We used those data to evaluate trend of AOD over almost fifty Indian cities having population more than a million. The cities we have chosen spread over almost entire length and breadth of the country. Few such studies have already been conducted using MODIS data. They typically used level 3 data. Since Level 3 data comes in 1x 1 degree gridded form they provide average value over a vast geographical region. We used level 2 dataset to enable us taking smaller region(1/2 x 1/2 degree here) centering the region of our interest . We used seasonal Mann-Kendall (M-K) statistics coupled with Sen's non-parametric slope estimation procedure to estimate monthwise and overall(i.e., yearly trend taking seasonality into account) AOD trend. We used median AOD for each month of every year to discard very high AOD's which we often get due to cloud contamination. Seasonal M-K test takes

  1. Evaluating Environmental Management Approaches to Alcohol and Other Drug Abuse Prevention. Prevention Updates

    ERIC Educational Resources Information Center

    DeJong, William; Langford, Linda M.

    2006-01-01

    Recent years have seen an upsurge in prevention work focused on changing the campus and community environments in which college students make decisions about alcohol and other drug (AOD) use. This approach, called "environmental management," is based on three fundamental premises: (1) Substance use problems are aggravated by a physical, social,…

  2. Community College Presidents' Role in Alcohol and Other Drug Abuse Prevention. Prevention Updates

    ERIC Educational Resources Information Center

    DeJong, William

    2006-01-01

    This publication is designed to guide presidents of community colleges and their administrative staff as they focus on alcohol and other drug (AOD) abuse prevention. The first section reviews the challenges facing community colleges in reducing student substance use and the strengths the colleges draw on in addressing it. The second section…

  3. Alcohol and Other Drugs on Campus: The Scope of the Problem. Infofacts/Resources

    ERIC Educational Resources Information Center

    Kapner, Daniel Ari

    2008-01-01

    The most widespread health problem on college and university campuses in the United States is high-risk alcohol and other drug (AOD) use. Recent reports confirm that the nation's campuses continue to encounter significant consequences as a result of this problem. This "Infofacts/Resources" offers an overview of the problem and highlights effective…

  4. Discovery and Characterization of a 5-Hydroxymethylfurfural Oxidase from Methylovorus sp. Strain MP688

    PubMed Central

    Dijkman, Willem P.

    2014-01-01

    In the search for useful and renewable chemical building blocks, 5-hydroxymethylfurfural (HMF) has emerged as a very promising candidate, as it can be prepared from sugars. HMF can be oxidized to 2,5-furandicarboxylic acid (FDCA), which is used as a substitute for petroleum-based terephthalate in polymer production. On the basis of a recently identified bacterial degradation pathway for HMF, candidate genes responsible for selective HMF oxidation have been identified. Heterologous expression of a protein from Methylovorus sp. strain MP688 in Escherichia coli and subsequent enzyme characterization showed that the respective gene indeed encodes an efficient HMF oxidase (HMFO). HMFO is a flavin adenine dinucleotide-containing oxidase and belongs to the glucose-methanol-choline-type flavoprotein oxidase family. Intriguingly, the activity of HMFO is not restricted to HMF, as it is active with a wide range of aromatic primary alcohols and aldehydes. The enzyme was shown to be relatively thermostable and active over a broad pH range. This makes HMFO a promising oxidative biocatalyst that can be used for the production of FDCA from HMF, a reaction involving both alcohol and aldehyde oxidations. PMID:24271187

  5. Depression, Anxiety, and Alcohol or Other Drug Use among College Students. Prevention Updates

    ERIC Educational Resources Information Center

    Ross, Virginia

    2004-01-01

    Community studies and reports from clinicians reveal that significant numbers of students on U.S. college campuses suffer from depression and/or anxiety and use alcohol or other drugs (AOD). Counselors in both the drug abuse and mental health fields confirm that students who seek mental health treatment often report symptoms of substance abuse,…

  6. Effects of cloud cover and meteorology in estimating ground-level air pollution using MAIAC AOD in the Yangtze River Delta

    NASA Astrophysics Data System (ADS)

    Xiao, Q.; Liu, Y.

    2017-12-01

    Satellite aerosol optical depth (AOD) has been used to assess fine particulate matter (PM2.5) pollution worldwide. However, non-random missing AOD due to cloud cover or high surface reflectance can cause up to 80% data loss and bias model-estimated spatial and temporal trends of PM2.5. Previous studies filled the data gap largely by spatial smoothing which ignored the impact of cloud cover and meteorology on aerosol loadings and has been shown to exhibit poor performance when monitoring stations are sparse or when there is seasonal large-scale missingness. Using the Yangtze River Delta of China as an example, we present a flexible Multiple Imputation (MI) method that combines cloud fraction, elevation, humidity, temperature, and spatiotemporal trends to impute the missing AOD. A two-stage statistical model driven by gap-filled AOD, meteorology and land use information was then fitted to estimate daily ground PM2.5 concentrations in 2013 and 2014 at 1 km resolution with complete coverage in space and time. The daily MI models have an average R2 of 0.77, with an inter-quartile range of 0.71 to 0.82 across days. The overall model 10-fold cross-validation R2 were 0.81 and 0.73 (for year 2013 and 2014, respectively. Predictions with only observational AOD or only imputed AOD showed similar accuracy. This method provides reliable PM2.5 predictions with complete coverage at high resolution. By including all the pixels of all days into model development, this method corrected the sampling bias in satellite-driven air pollution modelling due to non-random missingness in AOD. Comparing with previously reported gap-filling methods, the MI method has the strength of not relying on ground PM2.5 measurements, therefore allows the prediction of historical PM2.5 levels prior to the establishment of regular ground monitoring networks.

  7. Can the physical properties associated with uncertainties in the NASA MODIS AOD retrievals in the western U.S. be determined?

    NASA Astrophysics Data System (ADS)

    Loria Salazar, S. M.; Holmes, H.; Panorska, A. K.; Arnott, W. P.; Barnard, J.

    2016-12-01

    Previous investigations have used satellite remote sensing to estimate surface air pollution concentrations. While most of these studies rely on models developed for the dark-vegetated eastern U.S., they are being used in the semi-arid western U.S without modifications. These models are not robust in the western U.S. due to: 1. Irregular topography that leads to complicated boundary layer physics, 2. Pollutant mixtures, 3. Heterogeneous vertical profile of aerosol concentrations, and 4. High surface reflectance. Here, results from Nevada and California demonstrate poor AOD correlation between AERONET MODIS retrievals. Smoke from wildfires strengthened the aerosol signal, but the MODIS versus AERONET AOD correlation did not improve significantly during fire events [r2 0.17 (non-fire), r2 0.2 (fire)]. Furthermore, aerosol from fires increased the normalized mean bias (NMB) of MODIS retrievals of AOD[NMB 82% (non-fire), NMB 146% (fire)]. Additional results of this investigation found that aerosol plumes confined with the boundary layer improves MODIS AOD retrievals. However, when this condition is not met (i.e., 70% of the time downwind of mountains regions) MODIS AOD has a poor correlation and high bias with respect to AERONET AOD. Fire injection height, complicated boundary layer mixing, and entrainment disperse smoke plumes into the free atmosphere. Therefore, smoke plumes exacerbate the complex aerosol transport typical in the western U.S. and the non-linear relationship between surface pollutant concentrations and conditions aloft. This work uses stochastic methods, including regression to investigate the influence of each of these physical behaviors on the MODIS, AERONET AOD discrepancy using surrogates for each physical phenomenon, e.g., surface albedo for surface reflectance, boundary layer height and elevation for complex mixing, aerosol optical height for vertical aerosol concentrations, and fire radiative power for smoke plume injection height.

  8. Two decades of satellite observations of AOD over mainland China using ATSR-2, AATSR and MODIS/Terra: data set evaluation and large-scale patterns

    NASA Astrophysics Data System (ADS)

    de Leeuw, Gerrit; Sogacheva, Larisa; Rodriguez, Edith; Kourtidis, Konstantinos; Georgoulias, Aristeidis K.; Alexandri, Georgia; Amiridis, Vassilis; Proestakis, Emmanouil; Marinou, Eleni; Xue, Yong; van der A, Ronald

    2018-02-01

    The retrieval of aerosol properties from satellite observations provides their spatial distribution over a wide area in cloud-free conditions. As such, they complement ground-based measurements by providing information over sparsely instrumented areas, albeit that significant differences may exist in both the type of information obtained and the temporal information from satellite and ground-based observations. In this paper, information from different types of satellite-based instruments is used to provide a 3-D climatology of aerosol properties over mainland China, i.e., vertical profiles of extinction coefficients from the Cloud-Aerosol Lidar with Orthogonal Polarization (CALIOP), a lidar flying aboard the Cloud-Aerosol Lidar and Infrared Pathfinder Satellite Observation (CALIPSO) satellite and the column-integrated extinction (aerosol optical depth - AOD) available from three radiometers: the European Space Agency (ESA)'s Along-Track Scanning Radiometer version 2 (ATSR-2), Advanced Along-Track Scanning Radiometer (AATSR) (together referred to as ATSR) and NASA's Moderate Resolution Imaging Spectroradiometer (MODIS) aboard the Terra satellite, together spanning the period 1995-2015. AOD data are retrieved from ATSR using the ATSR dual view (ADV) v2.31 algorithm, while for MODIS Collection 6 (C6) the AOD data set is used that was obtained from merging the AODs obtained from the dark target (DT) and deep blue (DB) algorithms, further referred to as the DTDB merged AOD product. These data sets are validated and differences are compared using Aerosol Robotic Network (AERONET) version 2 L2.0 AOD data as reference. The results show that, over China, ATSR slightly underestimates the AOD and MODIS slightly overestimates the AOD. Consequently, ATSR AOD is overall lower than that from MODIS, and the difference increases with increasing AOD. The comparison also shows that neither of the ATSR and MODIS AOD data sets is better than the other one everywhere. However, ATSR ADV

  9. Electron and Fluorescence Microscopy of Extracellular Glucan and Aryl-Alcohol Oxidase during Wheat-Straw Degradation by Pleurotus eryngii

    PubMed Central

    Barrasa, J. M.; Gutiérrez, A.; Escaso, V.; Guillén, F.; Martínez, M. J.; Martínez, A. T.

    1998-01-01

    The ligninolytic fungus Pleurotus eryngii grown in liquid medium secreted extracellular polysaccharide (87% glucose) and the H2O2-producing enzyme aryl-alcohol oxidase (AAO). The production of both was stimulated by wheat-straw. Polyclonal antibodies against purified AAO were obtained, and a complex of glucanase and colloidal gold was prepared. With these tools, the localization of AAO and extracellular glucan in mycelium from liquid medium and straw degraded under solid-state fermentation conditions was investigated by transmission electron microscopy (TEM) and fluorescence microscopy. These studies revealed that P. eryngii produces a hyphal sheath consisting of a thin glucan layer. This sheath appeared to be involved in both mycelial adhesion to the straw cell wall during degradation and AAO immobilization on hyphal surfaces, with the latter evidenced by double labeling. AAO distribution during differential degradation of straw tissues was observed by immunofluorescence microscopy. Finally, TEM immunogold studies confirmed that AAO penetrates the plant cell wall during P. eryngii degradation of wheat straw. PMID:9435085

  10. The World Optical Depth Research and Calibration Center (WORCC) quality assurance and quality control of GAW-PFR AOD measurements

    NASA Astrophysics Data System (ADS)

    Kazadzis, Stelios; Kouremeti, Natalia; Nyeki, Stephan; Gröbner, Julian; Wehrli, Christoph

    2018-02-01

    The World Optical Depth Research Calibration Center (WORCC) is a section within the World Radiation Center at Physikalisches-Meteorologisches Observatorium (PMOD/WRC), Davos, Switzerland, established after the recommendations of the World Meteorological Organization for calibration of aerosol optical depth (AOD)-related Sun photometers. WORCC is mandated to develop new methods for instrument calibration, to initiate homogenization activities among different AOD networks and to run a network (GAW-PFR) of Sun photometers. In this work we describe the calibration hierarchy and methods used under WORCC and the basic procedures, tests and processing techniques in order to ensure the quality assurance and quality control of the AOD-retrieved data.

  11. Survey of Alcohol and Other Drug Use Attitudes and Behaviors in Nursing Students

    ERIC Educational Resources Information Center

    Baldwin, Jeffrey N.; Bartek, Jean K.; Scott, David M.; Davis-Hall, R. Ellen; DeSimone, Edward M., II

    2009-01-01

    Statewide nursing student alcohol and other drug (AOD) use attitudes and behaviors were assessed. Response was 929/2017 (46%) (practical nursing [n = 173/301] 57.3%; diploma and associate degree in nursing [n = 282/417] 67.6%; bachelor of science in nursing [n = 474/1299] 36.5%). Nearly 44% reported inadequate substance abuse education. Past-year…

  12. Neighborhood Racial and Socioeconomic Disparities in the Food and Alcohol Environment: Are There Differences by Commercial Data Sources?

    PubMed

    Mendez, Dara D; Kim, Kevin H; Hardaway, Cecily R; Fabio, Anthony

    2016-03-01

    This study examined neighborhood racial and socioeconomic disparities and the density of food and alcohol establishments. We also examined whether these disparities differed by data source. This study included commercial data for 2003 and 2009 from InfoUSA and Dun and Bradstreet (D&B) in 416 census tracts in Allegheny County, PA. Food and alcohol establishment densities were calculated by using area and population data from the 2000 US census. Differences between InfoUSA and D&B of food and alcohol densities across neighborhood racial and socioeconomic characteristics were tested using correlations and two-way mixed analysis of variance (ANOVA). There were differences by data source in the association between neighborhood racial and socioeconomic characteristics and food/alcohol establishment density. There was a positive correlation between grocery store/supermarket density and percentage black, poverty, and percentage without a car among D&B data but not in InfoUSA. Alcohol outlet density (AOD) increased as neighborhood poverty increased for both data sources, but the mean difference in AOD between InfoUSA and D&B was highest among neighborhoods with 25-50 % poverty (Cohen's d -0.49, p < 0.001) compared to neighborhoods with lower or higher poverty (2003 data). Mean grocery store density increased as percentage poverty increased, but only among D&B (2009 data). Differences in commercial data in the location and numeration of food and alcohol establishments are associated with neighborhood racial and socioeconomic characteristics and may introduce biases concerning neighborhood food and alcohol environments, racial and socioeconomic disparities, and health.

  13. Sharing stories: indigenous alcohol and other drug workers' well-being, stress and burnout.

    PubMed

    Roche, Ann M; Duraisingam, Vinita; Trifonoff, Allan; Battams, Samantha; Freeman, Toby; Tovell, Amanda; Weetra, Donna; Bates, Nancy

    2013-09-01

    Indigenous alcohol and other drug (AOD) workers' roles are often exhausting, poorly paid and under-recognised. There has been relatively little examination of work-related stressors on their health and well-being. This national study identified Indigenous AOD workers' experiences and perspectives on well-being, stress and burnout along with strategies to improve worker well-being. Focus groups were conducted with 121 participants (70 Indigenous, 20 non-Indigenous, 31 unspecified) from metropolitan, rural and remote locations around Australia, selected via a purposive sampling strategy. Audio files and interview notes were analysed to identify key themes. Main themes identified included excessive workloads, extensive demands and expectations, workers' proximity to communities, loss and grief issues, lack of recognition, inadequate rewards, stigma and racism, and Indigenous ways of working. Stressors were compounded by workers' complex personal circumstances, profound levels of loss and grief, and lack of culturally safe working environments. Indigenous workers' stress was exacerbated by close links and responsibilities to their communities and a 'dual accountability', being constantly on call, playing multiple roles, complex personal and professional lives, and needing to interact with multiple agencies. Many Indigenous AOD workers had developed mechanisms to deal with work-related pressures and received valued support from their communities. The study identified the importance of workforce strategies to improve Indigenous workers' well-being and reduce stress, including: mutual support networks, training in assertiveness and boundary setting, workloads that take account of Indigenous ways of working, adequate remuneration, supervision and mentorship, and cultural sensitivity training for non-Indigenous workers. © 2013 Australasian Professional Society on Alcohol and other Drugs.

  14. Profile of young people attending alcohol and other drug treatment services in Aotearoa, New Zealand: clinical file search.

    PubMed

    Schroder, Ria; Sellman, Doug; Frampton, Chris; Deering, Daryle

    2008-11-01

    The aim of the present study was to provide a profile of young people attending alcohol and other drug (AOD) treatment services in Aotearoa, New Zealand. Data were gathered from a clinical file search of 184 randomly selected young people aged 13-19 years who had attended one of eight youth AOD treatment services in New Zealand during 2003 or 2004. These services represented eight of the 11 youth-specific AOD services available to youth in New Zealand. Young people who attend youth-specific AOD services in New Zealand present with a range of complex needs including substance use and mental health issues, criminality, family conflict and disengagement from school. A total of 62.0% were male, 56.4% had criminal convictions, 40.6% had spent some time in Child, Youth and Family Services care and 53.8% were reported to have a coexisting substance use and mental health disorder. Low rates of reporting of substance use and mental health diagnoses in treatment files suggest that substance use and mental health disorders among this population are likely to be higher than those reported. This paper provides a unique profile of young people attending youth-specific AOD treatments in New Zealand. Such information is useful in informing treatment planning and funding and ensuring that service development occurs to specifically meet the complex needs of this patient group.

  15. Prevention of melanin formation during aryl alcohol oxidase production under growth-limited conditions using an Aspergillus nidulans cell factory.

    PubMed

    Pardo-Planas, Oscar; Prade, Rolf A; Müller, Michael; Atiyeh, Hasan K; Wilkins, Mark R

    2017-11-01

    An Aspergillus nidulans cell factory was genetically engineered to produce an aryl alcohol oxidase (AAO). The cell factory initiated production of melanin when growth-limited conditions were established using stationary plates and shaken flasks. This phenomenon was more pronounced when the strain was cultured in a trickle bed reactor (TBR). This study investigated different approaches to reduce melanin formation in fungal mycelia and liquid medium in order to increase the enzyme production yield. Removal of copper from the medium recipe reduced melanin formation in agar cultures and increased enzyme activities by 48% in agitated liquid cultures. Copper has been reported as a key element for tyrosinase, an enzyme responsible for melanin production. Ascorbic acid (0.44g/L) stopped melanin accumulation, did not affect growth parameters and resulted in AAO activity that was more than two-fold greater than a control treatment with no ascorbic acid. Copyright © 2017 Elsevier Ltd. All rights reserved.

  16. A Hepatocyte-Mimicking Antidote for Alcohol Intoxication.

    PubMed

    Xu, Duo; Han, Hui; He, Yuxin; Lee, Harrison; Wu, Di; Liu, Fang; Liu, Xiangsheng; Liu, Yang; Lu, Yunfeng; Ji, Cheng

    2018-04-11

    Alcohol intoxication causes serious diseases, whereas current treatments are mostly supportive and unable to remove alcohol efficiently. Upon alcohol consumption, alcohol is sequentially oxidized to acetaldehyde and acetate by the endogenous alcohol dehydrogenase and aldehyde dehydrogenase, respectively. Inspired by the metabolism of alcohol, a hepatocyte-mimicking antidote for alcohol intoxication through the codelivery of the nanocapsules of alcohol oxidase (AOx), catalase (CAT), and aldehyde dehydrogenase (ALDH) to the liver, where AOx and CAT catalyze the oxidation of alcohol to acetaldehyde, while ALDH catalyzes the oxidation of acetaldehyde to acetate. Administered to alcohol-intoxicated mice, the antidote rapidly accumulates in the liver and enables a significant reduction of the blood alcohol concentration. Moreover, blood acetaldehyde concentration is maintained at an extremely low level, significantly contributing to liver protection. Such an antidote, which can eliminate alcohol and acetaldehyde simultaneously, holds great promise for the treatment of alcohol intoxication and poisoning and can provide therapeutic benefits. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  17. Trends in Adolescent Alcohol and Other Drug Use: Findings from Three Sentinel Sites in South Africa (1997-2001)

    ERIC Educational Resources Information Center

    Parry, Charles D. H.; Myers, Bronwyn; Morojele, Neo K.; Flisher, Alan J.; Bhana, Arvin; Donson, Hilton; Pluddemann, Andreas

    2004-01-01

    This paper aims to provide surveillance information about the extent and consequences of alcohol and other drug (AOD) use by adolescents for three sentinel sites in South Africa (Cape Town, Durban and Gauteng province). From 1997 to 2001, data were gathered from multiple sources, including specialist treatment centres, trauma units, school…

  18. Children at the Front: A Different View of the War on Alcohol and Drugs. The CWLA North American Commission on Chemical Dependency and Child Welfare. Final Report and Recommendations.

    ERIC Educational Resources Information Center

    Child Welfare League of America, Inc., Washington, DC.

    This report explores the impact of alcohol and drug problems on the children and families served by the child welfare system and outlines a comprehensive strategy to better respond to their needs. It contains more than 100 policy and action recommendations to enhance services to children and families affected by alcohol and other drugs (AOD), to…

  19. Estimating Ground-Level Particulate Matter (PM) Concentration using Satellite-derived Aerosol Optical Depth (AOD)

    NASA Astrophysics Data System (ADS)

    Park, Seohui; Im, Jungho

    2017-04-01

    Atmospheric aerosols are strongly associated with adverse human health effects. In particular, particulate matter less than 10 micrometers and 2.5 micrometers (i.e., PM10 and PM2.5, respectively) can cause cardiovascular and lung diseases such as asthma and chronic obstructive pulmonary disease (COPD). Air quality including PM has typically been monitored using station-based in-situ measurements over the world. However, in situ measurements do not provide spatial continuity over large areas. An alternative approach is to use satellite remote sensing as it provides data over vast areas at high temporal resolution. The literature shows that PM concentrations are related with Aerosol Optical Depth (AOD) that is derived from satellite observations, but it is still difficult to identify PM concentrations directly from AOD. Some studies used statistical approaches for estimating PM concentrations from AOD while some others combined numerical models and satellite-derived AOD. In this study, satellite-derived products were used to estimate ground PM concentrations based on machine learning over South Korea. Satellite-derived products include AOD from Geostationary Ocean Color Imager (GOCI), precipitation from Tropical Rainfall Measuring Mission (TRMM), soil moisture from AMSR-2, elevation from Shuttle Radar Topography Mission (SRTM), and land cover, land surface temperature and normalized difference vegetation index (NDVI) from Moderate Resolution Imaging Spectroradiometer (MODIS). PM concentrations data were collected from 318 stations. A statistical ordinary least squares (OLS) approach was also tested and compared with the machine learning approach (i.e., random forest). PM concentration was estimated during spring season (from March to May) in 2015 that typically shows high concentration of PM. The randomly selected 80% of data were used for model calibration and the remaining 20% were used for validation. The developed models were further tested for prediction of PM

  20. EFFECT OF QUALITY CHRONIC DISEASE MANAGEMENT FOR ALCOHOL AND DRUG DEPENDENCE ON ADDICTION OUTCOMES

    PubMed Central

    Kim, Theresa W.; Saitz, Richard; Cheng, Debbie M.; Winter, Michael R; Witas, Julie; Samet, Jeffrey H.

    2012-01-01

    We examinedthe effect ofthe quality of primary care-basedchronic disease management (CDM)for alcohol and/or other drug (AOD) dependenceonaddiction outcomes.We assessed qualityusing 1)avisit frequencybased measure and 2) a self-reported assessment measuring alignment with the chronic care model. The visit frequency based measure had no significant association with addiction outcomes. Theself-reported measure of care - when care was at a CDM clinic - was associated with lower drug addiction severity.The self-reported assessment of care from any healthcare source (CDM clinic or elsewhere)was associated with lower alcoholaddiction severity and abstinence.These findings suggest that high quality CDM for AOD dependence may improve addiction outcomes.Quality measuresbased upon alignment with the chronic care model may better capture features of effective CDM care than a visitfrequency measure. PMID:22840687

  1. Physical Modelling of the Effect of Slag and Top-Blowing on Mixing in the AOD Process

    NASA Astrophysics Data System (ADS)

    Haas, Tim; Visuri, Ville-Valtteri; Kärnä, Aki; Isohookana, Erik; Sulasalmi, Petri; Eriç, Rauf Hürman; Pfeifer, Herbert; Fabritius, Timo

    The argon-oxygen decarburization (AOD) process is the most common process for refining stainless steel. High blowing rates and the resulting efficient mixing of the steel bath are characteristic of the AOD process. In this work, a 1:9-scale physical model was used to study mixing in a 150 t AOD vessel. Water, air and rapeseed oil were used to represent steel, argon and slag, respectively, while the dynamic similarity with the actual converter was maintained using the modified Froude number and the momentum number. Employing sulfuric acid as a tracer, the mixing times were determined on the basis of pH measurements according to the 97.5% criterion. The gas blowing rate and slag-steel volume ratio were varied in order to study their effect on the mixing time. The effect of top-blowing was also investigated. The results suggest that mixing time decreases as the modified Froude number of the tuyères increases and that the presence of a slag layer increases the mixing time. Furthermore, top-blowing was found to increase the mixing time both with and without the slag layer.

  2. Links between Alcohol and Other Drug Problems and Maltreatment among Adolescent Girls: Perceived Discrimination, Ethnic Identity, and Ethnic Orientation as Moderators

    ERIC Educational Resources Information Center

    Gray, Calonie M. K.; Montgomery, Marilyn J.

    2012-01-01

    Objectives: This study examined the links between maltreatment, posttraumatic stress symptoms, ethnicity-specific factors (i.e., perceived discrimination, ethnic identity, and ethnic orientation), and alcohol and/or other drug (AOD) problems among adolescent girls. Methods: These relations were examined using archived data from a community sample…

  3. Gender-specific HIV prevention interventions for women who use alcohol and other drugs: The evolution of the science and future directions

    PubMed Central

    Wechsberg, Wendee M.; Deren, Sherry; Myers, Bronwyn; Kirtadze, Irma; Zule, William A.; Howard, Brittni; El-Bassel, Nabila

    2015-01-01

    The use of alcohol and other drugs (AODs) is an important driver of gender disparities in HIV prevalence. Consequently, there is a need for women-specific HIV interventions that are conceptualized to address (1) women’s risk behavior, their roles in sexual relationships, and gender power dynamics, and (2) other issues commonly faced by women who use AODs, such as gender-based violence and victimization. This article presents the evolution of HIV prevention intervention research with women who use AODs. It looks at three generations of women-focused HIV research interventions, including first-generation projects that started in the 1990s, second-generation efforts where projects expanded in scope and included adaptions of evidence-based interventions for global relevance, and finally third-generation projects currently underway that combine biobehavioral methods and are being implemented in real-world settings. Because women who use AODs continue to report risk behaviors related to HIV, emphasis should be placed on training scientists to conduct gender-specific studies, increasing funding for new studies, and advocating to ensure that stigma-free services are available for these at-risk women. PMID:25978479

  4. Structural and kinetic studies on the Ser101Ala variant of choline oxidase: Catalysis by compromise

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

    Finnegan, S.; Orville, A.; Yuan, H.

    2010-09-15

    The oxidation of choline catalyzed by choline oxidase includes two reductive half-reactions where FAD is reduced by the alcohol substrate and by an aldehyde intermediate transiently formed in the reaction. Each reductive half-reaction is followed by an oxidative half-reaction where the reduced flavin is oxidized by oxygen. Here, we have used mutagenesis to prepare the Ser101Ala mutant of choline oxidase and have investigated the impact of this mutation on the structural and kinetic properties of the enzyme. The crystallographic structure of the Ser101Ala enzyme indicates that the only differences between the mutant and wild-type enzymes are the lack of amore » hydroxyl group on residue 101 and a more planar configuration of the flavin in the mutant enzyme. Kinetics established that replacement of Ser101 with alanine yields a mutant enzyme with increased efficiencies in the oxidative half-reactions and decreased efficiencies in the reductive half-reactions. This is accompanied by a significant decrease in the overall rate of turnover with choline. Thus, this mutation has revealed the importance of a specific residue for the optimization of the overall turnover of choline oxidase, which requires fine-tuning of four consecutive half-reactions for the conversion of an alcohol to a carboxylic acid.« less

  5. Assessment of the relationship between satellite AOD and ground PM₁₀ measurement data considering synoptic meteorological patterns and Lidar data.

    PubMed

    Zeeshan, Muhammad; Kim Oanh, N T

    2014-03-01

    Correlation between satellite aerosol optical depth (AOD) and ground monitoring particulate matter (PM) depends on the meteorology that determines PM optical properties, its dispersion, accumulation and vertical distribution. This study presents a novel approach to analyze PM-AOD relationship considering the totality of meteorological factors expressed as synoptic patterns. Meteorological observations at 07:00 Bangkok time from 9 regional meteorological stations, in dry seasons (November-April) of 11 years (2000-2010), were used to categorize governing meteorology over Central Thailand into four categories representing the typical observed synoptic patterns. The MANOVA analysis showed that these patterns were statistically different. PM10 recorded at 22 air quality stations in Bangkok Metropolitan Region were examined which showed the highest levels for the days belonging to pattern 1, followed by pattern 4, both with presence of a high pressure ridge, while the minimum for pattern 2 when thermal lows dominated. Lidar aerosol backscatter profiles recorded at Pimai station were used as indicator of PM vertical distribution that showed similarity within each pattern. R(2) between MODIS and Sun photometer AODs at Pimai was above 0.8. Correlation coefficients (R) between MODIS AOD and corresponding 1h PM10 for clear sky days (cloudiness ≤ 3/10) were examined for each pattern in comparison with lump case. Significant improvements were observed for pattern 1, average R across 22 stations was 0.46 for Terra and 0.38 for Aqua AOD compared to lump case with R of 0.34 and 0.31, respectively. Comparable improvement was also observed for pattern 4. For pattern 2, R values were significantly reduced which may be caused by the deeper mixing layers and varying vertical profiles with overall low values of Lidar backscatter coefficients. Improved R values in pattern 1 and 4, which had highest PM10 in BMR, suggested a better potential of using MODIS AOD for PM10 monitoring with

  6. Full-Coverage High-Resolution Daily PM(sub 2.5) Estimation using MAIAC AOD in the Yangtze River Delta of China

    NASA Technical Reports Server (NTRS)

    Xiao, Qingyang; Wang, Yujie; Chang, Howard H.; Meng, Xia; Geng, Guannan; Lyapustin, Alexei Ivanovich; Liu, Yang

    2017-01-01

    Satellite aerosol optical depth (AOD) has been used to assess population exposure to fine particulate matter (PM (sub 2.5)). The emerging high-resolution satellite aerosol product, Multi-Angle Implementation of Atmospheric Correction(MAIAC), provides a valuable opportunity to characterize local-scale PM(sub 2.5) at 1-km resolution. However, non-random missing AOD due to cloud snow cover or high surface reflectance makes this task challenging. Previous studies filled the data gap by spatially interpolating neighboring PM(sub 2.5) measurements or predictions. This strategy ignored the effect of cloud cover on aerosol loadings and has been shown to exhibit poor performance when monitoring stations are sparse or when there is seasonal large-scale missngness. Using the Yangtze River Delta of China as an example, we present a Multiple Imputation (MI) method that combines the MAIAC high-resolution satellite retrievals with chemical transport model (CTM) simulations to fill missing AOD. A two-stage statistical model driven by gap-filled AOD, meteorology and land use information was then fitted to estimate daily ground PM(sub 2.5) concentrations in 2013 and 2014 at 1 km resolution with complete coverage in space and time. The daily MI models have an average R(exp 2) of 0.77, with an inter-quartile range of 0.71 to 0.82 across days. The overall Ml model 10-fold cross-validation R(exp 2) (root mean square error) were 0.81 (25 gm(exp 3)) and 0.73 (18 gm(exp 3)) for year 2013 and 2014, respectively. Predictions with only observational AOD or only imputed AOD showed similar accuracy.Comparing with previous gap-filling methods, our MI method presented in this study performed bette rwith higher coverage, higher accuracy, and the ability to fill missing PM(sub 2.5) predictions without ground PM(sub 2.5) measurements. This method can provide reliable PM(sub 2.5)predictions with complete coverage that can reduce biasin exposure assessment in air pollution and health studies.

  7. Spatio-Temporal Variations in the Associations between Hourly PM2.5 and Aerosol Optical Depth (AOD) from MODIS Sensors on Terra and Aqua*

    PubMed Central

    Kim, Minho; Zhang, Xingyou; Holt, James B.; Liu, Yang

    2015-01-01

    Recent studies have explored the relationship between aerosol optical depth (AOD) measurements by satellite sensors and concentrations of particulate matter with aerodynamic diameters less than 2.5 μm (PM2.5). However, relatively little is known about spatial and temporal patterns in this relationship across the contiguous United States. In this study, we investigated the relationship between US Environmental Protection Agency estimates of PM2.5 concentrations and Moderate Resolution Imaging Spectroradiometer (MODIS) AOD measurements provided by two NASA satellites (Terra and Aqua) across the contiguous United States during 2005. We found that the combined use of both satellite sensors provided more AOD coverage than the use of either satellite sensor alone, that the correlation between AOD measurements and PM2.5 concentrations varied substantially by geographic location, and that this correlation was stronger in the summer and fall than that in the winter and spring. PMID:26336576

  8. Integrating motivational interviewing and traditional practices to address alcohol and drug use among urban American Indian/Alaska Native youth

    PubMed Central

    Dickerson, Daniel L.; Brown, Ryan A.; Johnson, Carrie L.; Schweigman, Kurt; D’Amico, Elizabeth J.

    2015-01-01

    American Indians/Alaska Natives (AI/AN) exhibit high levels of alcohol and drug (AOD) use and problems. Although approximately 70% of AI/ANs reside in urban areas, few culturally relevant AOD use programs targeting urban AI/AN youth exist. Furthermore, federally-funded studies focused on the integration of evidence-based treatments with AI/AN traditional practices are limited. The current study addresses a critical gap in the delivery of culturally appropriate AOD use programs for urban AI/AN youth, and outlines the development of a culturally tailored AOD program for urban AI/AN youth called Motivational Interviewing and Culture for Urban Native American Youth (MICUNAY). We conducted focus groups among urban AI/AN youth, providers, parents, and elders in two urban communities in northern and southern California aimed at 1) identifying challenges confronting urban AI/AN youth and 2) obtaining feedback on MICUNAY program content. Qualitative data were analyzed using Dedoose, a team-based qualitative and mixed methods analysis software platform. Findings highlight various challenges, including community stressors (e.g., gangs, violence), shortage of resources, cultural identity issues, and a high prevalence of AOD use within these urban communities. Regarding MICUNAY, urban AI/AN youth liked the collaborative nature of the motivational interviewing (MI) approach, especially with regard to eliciting their opinions and expressing their thoughts. Based on feedback from the youth, three AI/AN traditional practices (beading, AI/AN cooking, and prayer/sage ceremony) were chosen for the workshops. MICUNAY is the first AOD use prevention intervention program for urban AI/AN youth that integrates evidence-based treatment with traditional practices. This program addresses an important gap in services for this underserved population. PMID:26306776

  9. Alcohol consumption in the Australian coal mining industry.

    PubMed

    Tynan, Ross J; Considine, Robyn; Wiggers, John; Lewin, Terry J; James, Carole; Inder, Kerry; Kay-Lambkin, Frances; Baker, Amanda L; Skehan, Jaelea; Perkins, David; Kelly, Brian J

    2017-03-01

    To investigate patterns of alcohol use within the coal mining industry, and associations with the personal, social, workplace and employment characteristics. 8 mine sites across 3 eastern Australian states were surveyed, selected to encompass key geographic characteristics (accessibility and remoteness) and mine type (open cut and underground). Problematic alcohol use was measured using the Alcohol Use Disorders Identification Test (AUDIT) to determine: (1) overall risky or hazardous drinking behaviour; and (2) frequency of single-occasion drinking (6 or more drinks on 1 occasion). A total of 1457 employees completed the survey, of which 45.7% of male and 17.0% of female participants reported levels of alcohol use within the range considered as risky or hazardous, considerably higher than the national average. Hierarchical linear regression revealed a significant contribution of many individual level factors associated with AUDIT scores: younger age, male, current smoking status; illicit substance use; previous alcohol and other drug use (AOD) problems; and higher psychological distress. Workplace factors associated with alcohol use included working in mining primarily for the high remuneration, and the type of mining, with underground miners reporting higher alcohol use than open-cut miners. Our findings provide support for the need to address alcohol use in the coal mining industry over and above routine on-site testing for alcohol use. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://www.bmj.com/company/products-services/rights-and-licensing/.

  10. Estimating urban ground-level PM10 using MODIS 3km AOD product and meteorological parameters from WRF model

    NASA Astrophysics Data System (ADS)

    Ghotbi, Saba; Sotoudeheian, Saeed; Arhami, Mohammad

    2016-09-01

    Satellite remote sensing products of AOD from MODIS along with appropriate meteorological parameters were used to develop statistical models and estimate ground-level PM10. Most of previous studies obtained meteorological data from synoptic weather stations, with rather sparse spatial distribution, and used it along with 10 km AOD product to develop statistical models, applicable for PM variations in regional scale (resolution of ≥10 km). In the current study, meteorological parameters were simulated with 3 km resolution using WRF model and used along with the rather new 3 km AOD product (launched in 2014). The resulting PM statistical models were assessed for a polluted and largely variable urban area, Tehran, Iran. Despite the critical particulate pollution problem, very few PM studies were conducted in this area. The issue of rather poor direct PM-AOD associations existed, due to different factors such as variations in particles optical properties, in addition to bright background issue for satellite data, as the studied area located in the semi-arid areas of Middle East. Statistical approach of linear mixed effect (LME) was used, and three types of statistical models including single variable LME model (using AOD as independent variable) and multiple variables LME model by using meteorological data from two sources, WRF model and synoptic stations, were examined. Meteorological simulations were performed using a multiscale approach and creating an appropriate physic for the studied region, and the results showed rather good agreements with recordings of the synoptic stations. The single variable LME model was able to explain about 61%-73% of daily PM10 variations, reflecting a rather acceptable performance. Statistical models performance improved through using multivariable LME and incorporating meteorological data as auxiliary variables, particularly by using fine resolution outputs from WRF (R2 = 0.73-0.81). In addition, rather fine resolution for PM

  11. Patterns of Alcohol and Other Drug Use Associated with Major Depression among Gay Men Attending General Practices in Australia

    ERIC Educational Resources Information Center

    Holt, Martin; Bryant, Joanne; Newman, Christy E.; Paquette, Dana M.; Mao, Limin; Kidd, Michael R.; Saltman, Deborah C.; Kippax, Susan C.

    2012-01-01

    Our aim was to clarify the role of alcohol and other drug (AOD) use in major depression among gay men attending general medical practices. A secondary analysis was conducted on survey data collected from 531 gay men attending high-HIV-caseload general practices in Adelaide and Sydney, Australia. The survey contained demographic, social,…

  12. Existence of a novel enzyme, pyrroloquinoline quinone-dependent polyvinyl alcohol dehydrogenase, in a bacterial symbiont, Pseudomonas sp. strain VM15C.

    PubMed Central

    Shimao, M; Ninomiya, K; Kuno, O; Kato, N; Sakazawa, C

    1986-01-01

    A novel enzyme, pyrroloquinoline quinone (PQQ)-dependent polyvinyl alcohol (PVA) dehydrogenase, was found in and partially purified from the membrane fraction of a PVA-degrading symbiont, Pseudomonas sp. strain VM15C. The enzyme required PQQ for PVA dehydrogenation with phenazine methosulfate, phenazine ethosulfate, and 2,6-dichlorophenolindophenol as electron acceptors and did not show PVA oxidase activity leading to H2O2 formation. The enzyme was active toward low-molecular-weight secondary alcohols rather than primary alcohols. A membrane-bound PVA oxidase was also present in cells of VM15C. Although the purified oxidase showed a substrate specificity similar to that of PQQ-dependent PVA dehydrogenase and about threefold-higher PVA-dehydrogenating activity with phenazine methosulfate or phenazine ethosulfate than PVA oxidase activity with H2O2 formation, it was shown that the enzyme does not contain PQQ as the coenzyme, and PQQ did not affect its activity. Incubation of the membrane fraction of cells with PVA caused a reduction in the cytochrome(s) of the fraction. Images PMID:3513704

  13. Substance use by same sex attracted young people: Prevalence, perceptions and homophobia.

    PubMed

    Kelly, John; Davis, Cassandra; Schlesinger, Carla

    2015-07-01

    Research highlights that lesbian, gay, bisexual and transgender (LGBT) people use alcohol and drugs (AOD) more than heterosexual people; however, the incidence of AOD use by LGBT youth is less understood. The purpose of the current study was to ascertain AOD prevalence rates for LGBT youth compared with the Australian youth population; perceptions of AOD use within the LGBT community; and the impact of homophobia on AOD use. The study surveyed a cross-sectional sample of LGBT youth (13-24 years) (n = 161) who attended a LGBT festival in Brisbane, Australia, in 2012. The Alcohol Use Disorders Identification Test-Consumption, Fagerström Test for Nicotine Dependence and Drug Check Assessment Tool were utilised to examine patterns of AOD use, with items developed to explore perceptions of AOD use and homophobia. AOD use was common among the LGBT sample, with higher prevalence rates compared with the general Australian youth population (2010 National Drug Strategy Household Survey). AOD use by under 18-year-olds, and gender diverse youth was markedly higher. The majority misperceived AOD use to be the same in the LGBT and heterosexual communities. Those who believed homophobia impacted on AOD use were significantly more likely to use AOD. The higher prevalence of AOD use strongly suggests the need for AOD agencies to better respond to LGBT youth by not only screening sexuality and gender identity but also exploring young people's perceptions of AOD use in the LGBT community and their experiences of homophobia in order to provide effective AOD clinical treatment. © 2014 Australasian Professional Society on Alcohol and other Drugs.

  14. Correlations between Preoperative Angle Parameters and Postoperative Unpredicted Refractive Errors after Cataract Surgery in Open Angle Glaucoma (AOD 500).

    PubMed

    Lee, Wonseok; Bae, Hyoung Won; Lee, Si Hyung; Kim, Chan Yun; Seong, Gong Je

    2017-03-01

    To assess the accuracy of intraocular lens (IOL) power prediction for cataract surgery with open angle glaucoma (OAG) and to identify preoperative angle parameters correlated with postoperative unpredicted refractive errors. This study comprised 45 eyes from 45 OAG subjects and 63 eyes from 63 non-glaucomatous cataract subjects (controls). We investigated differences in preoperative predicted refractive errors and postoperative refractive errors for each group. Preoperative predicted refractive errors were obtained by biometry (IOL-master) and compared to postoperative refractive errors measured by auto-refractometer 2 months postoperatively. Anterior angle parameters were determined using swept source optical coherence tomography. We investigated correlations between preoperative angle parameters [angle open distance (AOD); trabecular iris surface area (TISA); angle recess area (ARA); trabecular iris angle (TIA)] and postoperative unpredicted refractive errors. In patients with OAG, significant differences were noted between preoperative predicted and postoperative real refractive errors, with more myopia than predicted. No significant differences were recorded in controls. Angle parameters (AOD, ARA, TISA, and TIA) at the superior and inferior quadrant were significantly correlated with differences between predicted and postoperative refractive errors in OAG patients (-0.321 to -0.408, p<0.05). Superior quadrant AOD 500 was significantly correlated with postoperative refractive differences in multivariate linear regression analysis (β=-2.925, R²=0.404). Clinically unpredicted refractive errors after cataract surgery were more common in OAG than in controls. Certain preoperative angle parameters, especially AOD 500 at the superior quadrant, were significantly correlated with these unpredicted errors.

  15. Integrating Motivational Interviewing and Traditional Practices to Address Alcohol and Drug Use Among Urban American Indian/Alaska Native Youth.

    PubMed

    Dickerson, Daniel L; Brown, Ryan A; Johnson, Carrie L; Schweigman, Kurt; D'Amico, Elizabeth J

    2016-06-01

    American Indians/Alaska Natives (AI/AN) exhibit high levels of alcohol and drug (AOD) use and problems. Although approximately 70% of AI/ANs reside in urban areas, few culturally relevant AOD use programs targeting urban AI/AN youth exist. Furthermore, federally-funded studies focused on the integration of evidence-based treatments with AI/AN traditional practices are limited. The current study addresses a critical gap in the delivery of culturally appropriate AOD use programs for urban AI/AN youth, and outlines the development of a culturally tailored AOD program for urban AI/AN youth called Motivational Interviewing and Culture for Urban Native American Youth (MICUNAY). We conducted focus groups among urban AI/AN youth, providers, parents, and elders in two urban communities in northern and southern California aimed at 1) identifying challenges confronting urban AI/AN youth and 2) obtaining feedback on MICUNAY program content. Qualitative data were analyzed using Dedoose, a team-based qualitative and mixed methods analysis software platform. Findings highlight various challenges, including community stressors (e.g., gangs, violence), shortage of resources, cultural identity issues, and a high prevalence of AOD use within these urban communities. Regarding MICUNAY, urban AI/AN youth liked the collaborative nature of the motivational interviewing (MI) approach, especially with regard to eliciting their opinions and expressing their thoughts. Based on feedback from the youth, three AI/AN traditional practices (beading, AI/AN cooking, and prayer/sage ceremony) were chosen for the workshops. To our knowledge, MICUNAY is the first AOD use prevention intervention program for urban AI/AN youth that integrates evidence-based treatment with traditional practices. This program addresses an important gap in services for this underserved population. Copyright © 2015. Published by Elsevier Inc.

  16. A decade of infrared versus visible AOD analysis within the dust belt

    NASA Astrophysics Data System (ADS)

    Capelle, Virginie; Chédin, Alain; Pondrom, Marc; Crevoisier, Cyril; Armante, Raymond; Crépeau, Laurent; Scott, Noëlle

    2017-04-01

    Aerosols represent one of the dominant uncertainties in radiative forcing, partly because of their very high spatiotemporal variability, a still insufficient knowledge of their microphysical and optical properties, or of their vertical distribution. A better understanding and forecasting of their impact on climate therefore requires precise observations of dust emission and transport. Observations from space offer a good opportunity to follow, day by day and at high spatial resolution, dust evolution at global scale and over long time series. In this context, infrared observations, by allowing retrieving simultaneously dust optical depth (AOD) as well as the mean dust layer altitude, daytime and nighttime, over oceans and over continents, in particular over desert, appears highly complementary to observations in the visible. In this study, a decade of infrared observations (Metop-A/IASI and AIRS/AQUA) has been processed pixel by pixel, using a "Look-Up-Table" (LUT) physical approach. The retrieved infrared 10µm coarse-mode AOD is compared with the Spectral Deconvolution Algorithm (SDA) 500nm coarse mode AOD observed at 50 ground-based Aerosol RObotic NETwork (AERONET) sites located within the dust belt. Analyzing their brings into evidence an important geographical variability. Lowest values are found close to dust sources ( 0.45 for the Sahel or Arabian Peninsula, 0.6-0.7 for the Northern part of Africa or India), whereas the ratio increases for transported dust with values of 0.9-1 for the Caribbean and for the Mediterranean basin. This variability is interpreted as a marker of clays abundance, and might be linked to the dust particle illite to kaolinite ratio, a recognized tracer of dust sources and transport. More generally, it suggests that the difference between the radiative impact of dust aerosols in the visible and in the infrared depends on the type of particles observed. This highlights the importance of taking into account the specificity of the

  17. Two Methods to Derive Ground-level Concentrations of PM2.5 with Improved Accuracy in the North China, Calibrating MODIS AOD and CMAQ Model Predictions

    NASA Astrophysics Data System (ADS)

    Lyu, Baolei; Hu, Yongtao; Chang, Howard; Russell, Armistead; Bai, Yuqi

    2016-04-01

    Reliable and accurate characterizations of ground-level PM2.5 concentrations are essential to understand pollution sources and evaluate human exposures etc. Monitoring network could only provide direct point-level observations at limited locations. At the locations without monitors, there are generally two ways to estimate the pollution levels of PM2.5. One is observations of aerosol properties from the satellite-based remote sensing, such as Moderate Resolution Imaging Spectroradiometer (MODIS) aerosol optical depth (AOD). The other one is from deterministic atmospheric chemistry models, such as the Community Multi-Scale Air Quality Model (CMAQ). In this study, we used a statistical spatio-temporal downscaler to calibrate the two datasets to monitor observations to derive fine-scale ground-level concentrations of PM2.5 with improved accuracy. We treated both MODIS AOD and CMAQ model predictions as biased proxy estimations of PM2.5 pollution levels. The downscaler proposed a Bayesian framework to model the spatially and temporally varying coefficients of the two types of estimations in the linear regression setting, in order to correct biases. Especially for calibrating MODIS AOD, a city-specific linear model was established to fill the missing AOD values, and a novel interpolation-based variable, i.e. PM2.5 Spatial Interpolator, was introduced to account for the spatial dependence among grid cells. We selected the heavy polluted and populated North China as our study area, in a grid setting of 81×81 12-km cells. For the evaluation of calibration performance for retrieved MODIS AOD, the R2 was 0.61 by the full model with PM2.5 Spatial Interpolator being presented, and was 0.48 with PM2.5 Spatial Interpolator not being presented. The constructed AOD values effectively predicted PM2.5 concentrations under our model structure, with R2=0.78. For the evaluation of calibrated CMAQ predictions, the R2 was 0.51, a little less than that of calibrated AOD. Finally we

  18. Different but the Same? Exploring the Experiences of Young People Caring for a Parent with an Alcohol or Other Drug Issue

    ERIC Educational Resources Information Center

    Moore; Tim; McArthur; Morag; Noble-Carr, Debbie

    2011-01-01

    Although children of parents with an alcohol or other drug (AOD) issue appear to assume a range of caring responsibilities within their families they have, until recently, been excluded from the growing body of young-carer research, policy and practice. This is problematic, as this group may experience greater levels of social exclusion whilst…

  19. The use of glucose oxidase and catalase for the enzymatic reduction of the potential ethanol content in wine.

    PubMed

    Röcker, Jessica; Schmitt, Matthias; Pasch, Ludwig; Ebert, Kristin; Grossmann, Manfred

    2016-11-01

    Due to the increase of sugar levels in wine grapes as one of the impacts of climate change, alcohol reduction in wines becomes a major focus of interest. This study combines the use of glucose oxidase and catalase activities with the aim of rapid conversion of glucose into non-fermentable gluconic acid. The H2O2 hydrolysing activity of purified catalase is necessary in order to stabilize glucose oxidase activity. After establishing the adequate enzyme ratio, the procedure was applied in large-scale trials (16L- and 220L-scale) of which one was conducted in a winery under industrial wine making conditions. Both enzyme activity and wine flavour were clearly influenced by the obligatory aeration in the different trials. With the enzyme treatment an alcohol reduction of 2%vol. was achieved after 30h of aeration. However the enzyme treated wines were significantly more acidic and less typical. Copyright © 2016. Published by Elsevier Ltd.

  20. Correlations between Preoperative Angle Parameters and Postoperative Unpredicted Refractive Errors after Cataract Surgery in Open Angle Glaucoma (AOD 500)

    PubMed Central

    Lee, Wonseok; Bae, Hyoung Won; Lee, Si Hyung; Kim, Chan Yun

    2017-01-01

    Purpose To assess the accuracy of intraocular lens (IOL) power prediction for cataract surgery with open angle glaucoma (OAG) and to identify preoperative angle parameters correlated with postoperative unpredicted refractive errors. Materials and Methods This study comprised 45 eyes from 45 OAG subjects and 63 eyes from 63 non-glaucomatous cataract subjects (controls). We investigated differences in preoperative predicted refractive errors and postoperative refractive errors for each group. Preoperative predicted refractive errors were obtained by biometry (IOL-master) and compared to postoperative refractive errors measured by auto-refractometer 2 months postoperatively. Anterior angle parameters were determined using swept source optical coherence tomography. We investigated correlations between preoperative angle parameters [angle open distance (AOD); trabecular iris surface area (TISA); angle recess area (ARA); trabecular iris angle (TIA)] and postoperative unpredicted refractive errors. Results In patients with OAG, significant differences were noted between preoperative predicted and postoperative real refractive errors, with more myopia than predicted. No significant differences were recorded in controls. Angle parameters (AOD, ARA, TISA, and TIA) at the superior and inferior quadrant were significantly correlated with differences between predicted and postoperative refractive errors in OAG patients (-0.321 to -0.408, p<0.05). Superior quadrant AOD 500 was significantly correlated with postoperative refractive differences in multivariate linear regression analysis (β=-2.925, R2=0.404). Conclusion Clinically unpredicted refractive errors after cataract surgery were more common in OAG than in controls. Certain preoperative angle parameters, especially AOD 500 at the superior quadrant, were significantly correlated with these unpredicted errors. PMID:28120576

  1. Longitudinal family effects on substance use among an at-risk adolescent sample

    PubMed Central

    Ewing, Brett A; Osilla, Karen Chan; Pedersen, Eric R; Hunter, Sarah B; Miles, Jeremy NV; D’Amico, Elizabeth J

    2014-01-01

    Objective Adult and peer factors may influence whether adolescents use alcohol and other drugs (AOD). This longitudinal study examined the direct effects of adult monitoring, perceived adult AOD use, and cultural values on adolescent AOD use. Methods Participants were 193 at-risk adolescents referred to a California diversion program called Teen Court for a first-time AOD offense. We assessed youth reports of past 30 day AOD use (any alcohol use, heavy drinking, marijuana use), demographics, changes in parental monitoring and family values (from baseline to follow-up 180 days later), as well as family structure and perceived adult substance use at follow-up. Results Adolescents who reported that a significant adult in their life used marijuana were more likely to have increased days of drinking, heavy drinking, and marijuana use at follow-up. Higher levels of familism (importance the teen places on their family’s needs over their own needs) and being in a nuclear family served as protective factors for future alcohol use. Additionally, poor family management was associated with increased alcohol use and heavy drinking. Conclusion Findings highlight how family management and perceptions of adult marijuana use influence subsequent adolescent AOD use, and how an increase in familism over time is associated with a decrease in adolescent drinking. Tailoring interventions, by including the teen’s family and/or providing support to adults who use AOD may be crucial for improving interventions for adolescent AOD use. PMID:25452064

  2. A structure-based catalytic mechanism for the xanthine oxidase family of molybdenum enzymes.

    PubMed Central

    Huber, R; Hof, P; Duarte, R O; Moura, J J; Moura, I; Liu, M Y; LeGall, J; Hille, R; Archer, M; Romão, M J

    1996-01-01

    The crystal structure of the xanthine oxidase-related molybdenum-iron protein aldehyde oxido-reductase from the sulfate reducing anaerobic Gram-negative bacterium Desulfovibrio gigas (Mop) was analyzed in its desulfo-, sulfo-, oxidized, reduced, and alcohol-bound forms at 1.8-A resolution. In the sulfo-form the molybdenum molybdopterin cytosine dinucleotide cofactor has a dithiolene-bound fac-[Mo, = O, = S, ---(OH2)] substructure. Bound inhibitory isopropanol in the inner compartment of the substrate binding tunnel is a model for the Michaelis complex of the reaction with aldehydes (H-C = O,-R). The reaction is proposed to proceed by transfer of the molybdenum-bound water molecule as OH- after proton transfer to Glu-869 to the carbonyl carbon of the substrate in concert with hydride transfer to the sulfido group to generate [MoIV, = O, -SH, ---(O-C = O, -R)). Dissociation of the carboxylic acid product may be facilitated by transient binding of Glu-869 to the molybdenum. The metal-bound water is replenished from a chain of internal water molecules. A second alcohol binding site in the spacious outer compartment may cause the strong substrate inhibition observed. This compartment is the putative binding site of large inhibitors of xanthine oxidase. Images Fig. 1 Fig. 2 Fig. 3 Fig. 4 Fig. 5 Fig. 6 PMID:8799115

  3. Molecular characterization of the fatty alcohol oxidation pathway for wax-ester mobilization in germinated jojoba seeds

    USDA-ARS?s Scientific Manuscript database

    Jojoba (Simmondsia chinensis) is the only plant species known to use liquid wax esters (WE) as a primary seed storage reserve. Upon germination, WE hydrolysis releases very long-chain fatty alcohols, which must be oxidised to fatty acids by the sequential action of a fatty alcohol oxidase (FAO) and ...

  4. Rationally engineered flavin-dependent oxidase reveals steric control of dioxygen reduction.

    PubMed

    Zafred, Domen; Steiner, Barbara; Teufelberger, Andrea R; Hromic, Altijana; Karplus, P Andrew; Schofield, Christopher J; Wallner, Silvia; Macheroux, Peter

    2015-08-01

    The ability of flavoenzymes to reduce dioxygen varies greatly, and is controlled by the protein environment, which may cause either a rapid reaction (oxidases) or a sluggish reaction (dehydrogenases). Previously, a 'gatekeeper' amino acid residue was identified that controls the reactivity to dioxygen in proteins from the vanillyl alcohol oxidase superfamily of flavoenzymes. We have identified an alternative gatekeeper residue that similarly controls dioxygen reactivity in the grass pollen allergen Phl p 4, a member of this superfamily that has glucose dehydrogenase activity and the highest redox potential measured in a flavoenzyme. A substitution at the alternative gatekeeper site (I153V) transformed the enzyme into an efficient oxidase by increasing dioxygen reactivity by a factor of 60,000. An inverse exchange (V169I) in the structurally related berberine bridge enzyme (BBE) decreased its dioxygen reactivity by a factor of 500. Structural and biochemical characterization of these and additional variants showed that our model enzymes possess a cavity that binds an anion and resembles the 'oxyanion hole' in the proximity of the flavin ring. We showed also that steric control of access to this site is the most important parameter affecting dioxygen reactivity in BBE-like enzymes. Analysis of flavin-dependent oxidases from other superfamilies revealed similar structural features, suggesting that dioxygen reactivity may be governed by a common mechanistic principle. Structural data are available in PDB database under the accession numbers 4PVE, 4PVH, 4PVJ, 4PVK, 4PWB, 4PWC and 4PZF. © 2015 FEBS.

  5. AOD Distributions and Trends of Major Aerosol Species over a Selection of the World's Most Populated Cities Based on the 1st Version of NASA's MERRA Aerosol Reanalysis

    NASA Technical Reports Server (NTRS)

    Provencal, Simon; Kishcha, Pavel; da Silva, Arlindo M.; Elhacham, Emily; Alpert, Pinhas

    2017-01-01

    NASA recently extended the Modern-Era Retrospective Analysis for Research and Application (MERRA) with an atmospheric aerosol reanalysis which includes five particulate species: sulfate, organic matter, black carbon, mineral dust and sea salt. The MERRA Aerosol Reanalysis (MERRAero) is an innovative tool to study air quality issues around the world for its global and constant coverage and its distinction of aerosol speciation expressed in the form of aerosol optical depth (AOD). The purpose of this manuscript is to apply MERRAero to the study of urban air pollution at the global scale by analyzing the AOD over a period of 13 years (2003-2015) and over a selection of 200 of the world's most populated cities in order to assess the impacts of urbanization, industrialization, air quality regulations and regional transport which affect urban aerosol load. Environmental regulations and the recent global economic recession have helped to decrease the AOD and sulfate aerosols in most cities in North America, Europe and Japan. Rapid industrialization in China over the last two decades resulted in Chinese cities having the highest AOD values in the world. China has nevertheless recently implemented emission control measures which are showing early signs of success in many cities of Southern China where AOD has decreased substantially over the last 13 years. The AOD over South American cities, which is dominated by carbonaceous aerosols, has also decreased over the last decade due to an increase in commodity prices which slowed deforestation activities in the Amazon rainforest. At the opposite, recent urbanization and industrialization in India and Bangladesh resulted in a strong increase of AOD, sulfate and carbonaceous aerosols in most cities of these two countries. The AOD over most cities in Northern Africa and Western Asia changed little over the last decade. Emissions of natural aerosols, which cities in these two regions tend to be mostly composed of, don't tend to

  6. AOD distributions and trends of major aerosol species over a selection of the world's most populated cities based on the 1st Version of NASA's MERRA Aerosol Reanalysis.

    PubMed

    Provençal, Simon; Kishcha, Pavel; da Silva, Arlindo M; Elhacham, Emily; Alpert, Pinhas

    2017-06-01

    NASA recently extended the Modern-Era Retrospective Analysis for Research and Application (MERRA) with an atmospheric aerosol reanalysis which includes five particulate species: sulfate, organic matter, black carbon, mineral dust and sea salt. The MERRA Aerosol Reanalysis (MERRAero) is an innovative tool to study air quality issues around the world for its global and constant coverage and its distinction of aerosol speciation expressed in the form of aerosol optical depth (AOD). The purpose of this manuscript is to apply MERRAero to the study of urban air pollution at the global scale by analyzing the AOD over a period of 13 years (2003-2015) and over a selection of 200 of the world's most populated cities in order to assess the impacts of urbanization, industrialization, air quality regulations and regional transport which affect urban aerosol load. Environmental regulations and the recent global economic recession have helped to decrease the AOD and sulfate aerosols in most cities in North America, Europe and Japan. Rapid industrialization in China over the last two decades resulted in Chinese cities having the highest AOD values in the world. China has nevertheless recently implemented emission control measures which are showing early signs of success in many cities of Southern China where AOD has decreased substantially over the last 13 years. The AOD over South American cities, which is dominated by carbonaceous aerosols, has also decreased over the last decade due to an increase in commodity prices which slowed deforestation activities in the Amazon rainforest. At the opposite, recent urbanization and industrialization in India and Bangladesh resulted in a strong increase of AOD, sulfate and carbonaceous aerosols in most cities of these two countries. The AOD over most cities in Northern Africa and Western Asia changed little over the last decade. Emissions of natural aerosols, which cities in these two regions tend to be mostly composed of, don't tend to

  7. Neuroimmune Basis of Alcoholic Brain Damage

    PubMed Central

    Crews, Fulton T.; Vetreno, Ryan P.

    2017-01-01

    Alcohol-induced brain damage likely contributes to the dysfunctional poor decisions associated with alcohol dependence. Human alcoholics have a global loss of brain volume that is most severe in the frontal cortex. Neuroimmune gene induction by binge drinking increases neurodegeneration through increased oxidative stress, particularly NADPH oxidase-induced oxidative stress. In addition, HMGB1-TLR4 and innate immune NF-κB target genes are increased leading to persistent and sensitized neuroimmune responses to ethanol and other agents that release HMGB1 or directly stimulate TLR receptors and/or NMDA receptors. Neuroimmune signaling and glutamate excitotoxicity are linked to alcoholic neurodegeneration. Models of adolescent alcohol abuse lead to significant frontal cortical degeneration and show the most severe loss of hippocampal neurogenesis. Adolescence is a period of high risk for ethanol-induced neurodegeneration and alterations in brain structure, gene expression, and maturation of adult phenotypes. Together, these findings support the hypothesis that adolescence is a period of risk for persistent and long-lasting increases in brain neuroimmune gene expression that promote persistent and long-term increases in alcohol consumption, neuroimmune gene induction, and neurodegeneration that we find associated with alcohol use disorders. PMID:25175868

  8. Functional expression of aryl-alcohol oxidase in Saccharomyces cerevisiae and Pichia pastoris by directed evolution.

    PubMed

    Viña-Gonzalez, Javier; Elbl, Katarina; Ponte, Xavier; Valero, Francisco; Alcalde, Miguel

    2018-07-01

    Aryl-alcohol oxidase (AAO) plays a fundamental role in the fungal ligninolytic secretome, acting as a supplier of H 2 O 2 . Despite its highly selective mechanism of action, the presence of this flavooxidase in different biotechnological settings has hitherto been hampered by the lack of appropriate heterologous expression systems. We recently described the functional expression of the AAO from Pleurotus eryngii in Saccharomyces cerevisiae by fusing a chimeric signal peptide (preαproK) and applying structure-guided evolution. Here, we have obtained an AAO secretion variant that is readily expressed in S. cerevisiae and overproduced in Pichia pastoris. First, the functional expression of AAO in S. cerevisiae was enhanced through the in vivo shuffling of a panel of secretion variants, followed by the focused evolution of the preαproK peptide. The outcome of this evolutionary campaign-an expression variant that accumulated 4 mutations in the chimeric signal peptide, plus two mutations in the mature protein- showed 350-fold improved secretion (4.5 mg/L) and was stable. This secretion mutant was cloned into P. pastoris and fermented in a fed-batch bioreactor to enhance production to 25 mg/L. While both recombinant AAO from S. cerevisiae and P. pastoris were subjected to the same N-terminal processing and had a similar pH activity profile, they differed in their kinetic parameters and thermostability. The strong glycosylation observed in the evolved AAO from S. cerevisiae underpinned this effect, since when the mutant was produced in the glycosylation-deficient S. cerevisiae strain Δkre2, its kinetic parameters and thermostability were comparable to its poorly glycosylated P. pastoris recombinant counterpart. © 2018 Wiley Periodicals, Inc.

  9. Interaction between a functional MAOA locus and childhood sexual abuse predicts alcoholism and antisocial personality disorder in adult women.

    PubMed

    Ducci, F; Enoch, M-A; Hodgkinson, C; Xu, K; Catena, M; Robin, R W; Goldman, D

    2008-03-01

    Women who have experienced childhood sexual abuse (CSA) have an increased risk of alcoholism and antisocial personality disorder (ASPD). Among male subjects, a functional polymorphism (MAOA-LPR, monoamine oxidase A linked polymorphic region) in the promoter region of the monoamine oxidase A gene (MAOA) appears to moderate the effect of childhood maltreatment on antisocial behavior. Our aim was to test whether MAOA-LPR influences the impact of CSA on alcoholism and ASPD in a sample of 291 women, 50% of whom have experienced CSA; we also tested whether haplotypes covering the region where both MAOA and monoamine oxidase B (MAOB) genes are located predict risk of alcoholism and ASPD better than the MAOA-LPR locus alone. Participants included 168 alcoholics (39 with ASPD (antisocial alcoholics) and 123 controls (no alcoholics, no ASPD). Antisocial behavior was also modeled as a continuous trait: ASPD symptoms count. The MAOA-LPR low activity allele was associated with alcoholism (P=0.005), particularly antisocial alcoholism (P=0.00009), only among sexually abused subjects. Sexually abused women who were homozygous for the low activity allele had higher rates of alcoholism and ASPD, and more ASPD symptoms, than abused women homozygous for the high activity allele. Heterozygous women displayed an intermediate risk pattern. In contrast, there was no relationship between alcoholism/antisocial behavior and MAOA-LPR genotype among non-abused women. The MAOA-LPR low activity allele was found on three different haplotypes. The most abundant MAOA haplotype containing the MAOA-LPR low activity allele was found in excess among alcoholics (P=0.008) and antisocial alcoholics (P=0.001). Finally, a MAOB haplotype, which we termed haplotype C, was significantly associated with alcoholism (P=0.006), and to a lesser extent with antisocial alcoholism (P=0.03). In conclusions, MAOA seems to moderate the impact of childhood trauma on adult psychopathology in female subjects in the same way

  10. Paid protection? Ethics of incentivised long-acting reversible contraception in adolescents with alcohol and other drug use.

    PubMed

    Won, Tiana; Blumenthal-Barby, Jennifer; Chacko, Mariam

    2017-03-01

    Pregnant adolescents have a higher risk of poor maternal and fetal outcomes, particularly in the setting of concomitant maternal alcohol and other drug (AOD) use. Despite numerous programmes aimed at reducing overall teen pregnancy rates and the recognition of AOD use as a risk factor for unintended pregnancy in adolescents, interventions targeting this specific group have been sparse. In adult drug-using women, financial incentives for contraception have been provided but are ethically controversial. This article explores whether a trial could ethically employ monetary incentives in adolescents with AOD use to promote the use of long-acting reversible contraception (LARC), with special attention to the relevant distinctions between adults and adolescents. We conclude that a trial of incentives to promote LARC in this patient population is ethically permissible if the incentives are small, are tied to completion of an educational activity to minimise the quick fix temptation and potential for non-attendance to the risks and benefits of LARC and are provided only to the adolescent after an assessment of her reasoning to rule out coercion (eg, by guardians) as motivation. Information about treatment for AOD use and follow-up care in case of problems with the contraceptive or desire for removal should also be provided. Before implementing such a trial, qualitative research with input from providers, potential patients and their parents should be conducted to inform the programme's specific structure. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://www.bmj.com/company/products-services/rights-and-licensing/.

  11. AOD Screening Tools for College Students. Prevention Update

    ERIC Educational Resources Information Center

    Higher Education Center for Alcohol, Drug Abuse, and Violence Prevention, 2012

    2012-01-01

    According to the National Institute on Alcohol Abuse and Alcoholism (NIAAA), the goal of screening in student health or other college settings is to reduce alcohol-related harm. NIAAA points out that identifying those students at greatest risk for alcohol problems is the first step in prevention. Colleges and universities have used a number of…

  12. Mit1 Transcription Factor Mediates Methanol Signaling and Regulates the Alcohol Oxidase 1 (AOX1) Promoter in Pichia pastoris*

    PubMed Central

    Wang, Xiaolong; Wang, Qi; Wang, Jinjia; Bai, Peng; Shi, Lei; Shen, Wei; Zhou, Mian; Zhou, Xiangshan; Zhang, Yuanxing; Cai, Menghao

    2016-01-01

    The alcohol oxidase 1 (AOX1) promoter (PAOX1) of Pichia pastoris is the most powerful and commonly used promoter for driving protein expression. However, mechanisms regulating its transcriptional activity are unclear. Here, we identified a Zn(II)2Cys6-type methanol-induced transcription factor 1 (Mit1) and elucidated its roles in regulating PAOX1 activity in response to glycerol and methanol. Mit1 regulated the expression of many genes involved in methanol utilization pathway, including AOX1, but did not participate in peroxisome proliferation and transportation of peroxisomal proteins during methanol metabolism. Structural analysis of Mit1 by performing domain deletions confirmed its specific and critical role in the strict repression of PAOX1 in glycerol medium. Importantly, Mit1, Mxr1, and Prm1, which positively regulated PAOX1 in response to methanol, were bound to PAOX1 at different sites and did not interact with each other. However, these factors cooperatively activated PAOX1 through a cascade. Mxr1 mainly functioned during carbon derepression, whereas Mit1 and Prm1 functioned during methanol induction, with Prm1 transmitting methanol signal to Mit1 by binding to the MIT1 promoter (PMIT1), thus increasingly expressing Mit1 and subsequently activating PAOX1. PMID:26828066

  13. Independently recruited oxidases from the glucose-methanol-choline oxidoreductase family enabled chemical defences in leaf beetle larvae (subtribe Chrysomelina) to evolve

    PubMed Central

    Rahfeld, Peter; Kirsch, Roy; Kugel, Susann; Wielsch, Natalie; Stock, Magdalena; Groth, Marco; Boland, Wilhelm; Burse, Antje

    2014-01-01

    Larvae of the leaf beetle subtribe Chrysomelina sensu stricto repel their enemies by displaying glandular secretions that contain defensive compounds. These repellents can be produced either de novo (iridoids) or by using plant-derived precursors (e.g. salicylaldehyde). The autonomous production of iridoids, as in Phaedon cochleariae, is the ancestral chrysomeline chemical defence and predates the evolution of salicylaldehyde-based defence. Both biosynthesis strategies include an oxidative step of an alcohol intermediate. In salicylaldehyde-producing species, this step is catalysed by salicyl alcohol oxidases (SAOs) of the glucose-methanol-choline (GMC) oxidoreductase superfamily, but the enzyme oxidizing the iridoid precursor is unknown. Here, we show by in vitro as well as in vivo experiments that P. cochleariae also uses an oxidase from the GMC superfamily for defensive purposes. However, our phylogenetic analysis of chrysomeline GMC oxidoreductases revealed that the oxidase of the iridoid pathway originated from a GMC clade different from that of the SAOs. Thus, the evolution of a host-independent chemical defence followed by a shift to a host-dependent chemical defence in chrysomeline beetles coincided with the utilization of genes from different GMC subfamilies. These findings illustrate the importance of the GMC multi-gene family for adaptive processes in plant–insect interactions. PMID:24943369

  14. Improvement of aerosol optical properties modeling over Eastern Asia with MODIS AOD assimilation in a global non-hydrostatic icosahedral aerosol transport model.

    PubMed

    Dai, Tie; Schutgens, Nick A J; Goto, Daisuke; Shi, Guangyu; Nakajima, Teruyuki

    2014-12-01

    A new global aerosol assimilation system adopting a more complex icosahedral grid configuration is developed. Sensitivity tests for the assimilation system are performed utilizing satellite retrieved aerosol optical depth (AOD) from the Moderate Resolution Imaging Spectroradiometer (MODIS), and the results over Eastern Asia are analyzed. The assimilated results are validated through independent Aerosol Robotic Network (AERONET) observations. Our results reveal that the ensemble and local patch sizes have little effect on the assimilation performance, whereas the ensemble perturbation method has the largest effect. Assimilation leads to significantly positive effect on the simulated AOD field, improving agreement with all of the 12 AERONET sites over the Eastern Asia based on both the correlation coefficient and the root mean square difference (assimilation efficiency). Meanwhile, better agreement of the Ångström Exponent (AE) field is achieved for 8 of the 12 sites due to the assimilation of AOD only. Copyright © 2014 Elsevier Ltd. All rights reserved.

  15. A Mathematical Model for Reactions During Top-Blowing in the AOD Process: Validation and Results

    NASA Astrophysics Data System (ADS)

    Visuri, Ville-Valtteri; Järvinen, Mika; Kärnä, Aki; Sulasalmi, Petri; Heikkinen, Eetu-Pekka; Kupari, Pentti; Fabritius, Timo

    2017-06-01

    In earlier work, a fundamental mathematical model was proposed for side-blowing operation in the argon oxygen decarburization (AOD) process. In the preceding part "Derivation of the Model," a new mathematical model was proposed for reactions during top-blowing in the AOD process. In this model it was assumed that reactions occur simultaneously at the surface of the cavity caused by the gas jet and at the surface of the metal droplets ejected from the metal bath. This paper presents validation and preliminary results with twelve industrial heats. In the studied heats, the last combined-blowing stage was altered so that oxygen was introduced from the top lance only. Four heats were conducted using an oxygen-nitrogen mixture (1:1), while eight heats were conducted with pure oxygen. Simultaneously, nitrogen or argon gas was blown via tuyères in order to provide mixing that is comparable to regular practice. The measured carbon content varied from 0.4 to 0.5 wt pct before the studied stage to 0.1 to 0.2 wt pct after the studied stage. The results suggest that the model is capable of predicting changes in metal bath composition and temperature with a reasonably high degree of accuracy. The calculations indicate that the top slag may supply oxygen for decarburization during top-blowing. Furthermore, it is postulated that the metal droplets generated by the shear stress of top-blowing create a large mass exchange area, which plays an important role in enabling the high decarburization rates observed during top-blowing in the AOD process. The overall rate of decarburization attributable to top-blowing in the last combined-blowing stage was found to be limited by the mass transfer of dissolved carbon.

  16. Drug and alcohol-impaired driving among electronic music dance event attendees.

    PubMed

    Furr-Holden, Debra; Voas, Robert B; Kelley-Baker, Tara; Miller, Brenda

    2006-10-15

    Drug-impaired driving has received increased attention resulting from development of rapid drug-screening procedures used by police and state laws establishing per se limits for drug levels in drivers. Venues that host electronic music dance events (EMDEs) provide a unique opportunity to assess drug-impaired driving among a high proportion of young adult drug users. EMDEs are late-night dance parties marked by a substantial number of young adult attendees and elevated drug involvement. No studies to date have examined drug-impaired driving in a natural environment with active drug and alcohol users. Six EMDEs were sampled in San Diego, California, and Baltimore, Maryland. A random sample of approximately 40 attendees per event were administered surveys about alcohol and other drug (AOD) use and driving status, given breath tests for alcohol, and asked to provide oral fluid samples to test for illicit drug use upon entering and exiting the events. Driving status reduced the level of alcohol use (including abstaining) but the impact on drug-taking was not significant. However, 62% of individuals who reported their intention to drive away from the events were positive for drugs or alcohol upon leaving. This suggests that these events and settings are appropriate ones for developing interventions for reducing risks for young adults.

  17. Culturally sensitive substance abuse intervention for Hispanic and African American adolescents: empirical examples from the Alcohol Treatment Targeting Adolescents in Need (ATTAIN) Project.

    PubMed

    Gil, Andrés G; Wagner, Eric F; Tubman, Jonathan G

    2004-11-01

    This study presents preliminary analyses examining the effects of an alcohol and other drug use (AOD) intervention with minority juvenile offenders. Furthermore, the study investigates the impact of cultural factors on baseline AOD use among Hispanic and African American youth, as well as on treatment outcome. Participants were 213 juvenile offenders referred for treatment (mean age = 15.7 years), 97 of whom have completed treatment to date. The intervention was carried out in clinics placed within the neighborhoods in which the participants resided. Intervention Alcohol Treatment Targeting Adolescents in Need (ATTAIN) is a controlled clinical trial evaluating the effectiveness of a brief motivational, cognitive behavioral intervention, guided self-change (GSC). Participants are assigned randomly to the individual format of guided self-change (I-GSC), the family involved format of guided self-Change (F-GSC), choice of one of these two, or a waiting list control condition. Only participants involved in active intervention are included in the present report. Data were collected via structured face-to-face interviews. Alcohol and marijuana use measures were collected using the Time-line Follow-back interview (TLFB). There were significant reductions in alcohol and marijuana use for all ethnic groups from baseline to post-intervention. Cultural factors (discrimination, acculturation, ethnic pride and cultural mistrust) were associated with pre-intervention levels of alcohol and marijuana use. Among Hispanics, pre-intervention level of substance use were higher among foreign-born than US-born youth. Analyses conducted with the US-born Hispanic group showed that ethnic orientation and ethnic pride were associated positively with greater reductions in alcohol use. The intervention provided through ATTAIN appears to be effective with a multi-ethnic population of juvenile delinquents. Cultural factors, such as ethnic orientation and ethnic mistrust, appear to constitute

  18. How online counselling can support partners of individuals with problem alcohol or other drug use.

    PubMed

    Wilson, Samara R; Rodda, Simone; Lubman, Dan I; Manning, Victoria; Yap, Marie B H

    2017-07-01

    Problematic alcohol and other drug (AOD) use impacts partners heavily, with an increased risk of experiencing domestic violence, financial stressors, health problems and relationship challenges. However, partners often do not seek help or support due to a range of barriers (e.g., shame, stigma, practical constraints). Online counselling may facilitate help-seeking by overcoming many of these barriers, however research is needed to explore what motivates partners to contact online counselling services, their experiences and needs, and how partners can be best supported online. One hundred transcripts of partners of individuals with problem AOD use were sampled from a 24-hour national AOD synchronous online chat counselling service. Descriptive content analysis was used to investigate themes related to help-seeking. Three broad themes, with seven sub-themes, were identified: (i) the reason for accessing online counselling (seeking advice, wanting to talk), (ii) discussing help-seeking and coping processes (past/present help-seeking or coping strategies, barriers and facilitators to seeking help and change), and (iii) planning for future assistance (future planning, treatment preferences). Partners wanted to talk about their concerns with a non-judgemental professional. However, the majority of help-seekers wanted advice and assistance in problem-solving, coping and the process of seeking further help. Future studies need to examine the impact of online help-seeking by partners. Copyright © 2017 Elsevier Inc. All rights reserved.

  19. Identifying Optimal Temporal Scale for the Correlation of AOD and Ground Measurements of PM2.5 to Improve the Modeling Performance in a Real-Time Air Quality Estimation System

    NASA Technical Reports Server (NTRS)

    Li,Hui; Faruque, Fazlay; Williams, Worth; Al-Hamdan, Mohammad; Luvall, Jeffrey; Crosson, William; Rickman, Douglas; Limaye, Ashutosh

    2008-01-01

    Aerosol optical depth (AOD), derived from satellite measurements using Moderate Resolution Imaging Spectrometer (MODIS), offers indirect estimates of particle matter. Research shows a significant positive correlation between satellite-based measurements of AOD and ground-based measurements of particulate matter with aerodynamic diameter less than or equal to 2.5 micrometers (PM2.5). In addition, satellite observations have also shown great promise in improving estimates of PM2.5 air quality surface. Research shows that correlations between AOD and ground PM2.5 are affected by a combination of many factors such as inherent characteristics of satellite observations, terrain, cloud cover, height of the mixing layer, and weather conditions, and thus might vary widely in different regions, different seasons, and even different days in a same location. Analysis of correlating AOD with ground measured PM2.5 on a day-to-day basis suggests the temporal scale, a number of immediate latest days for a given run's day, for their correlations needs to be considered to improve air quality surface estimates, especially when satellite observations are used in a real-time pollution system. The second reason is that correlation coefficients between AOD and ground PM2.5 cannot be predetermined and needs to be calculated for each day's run for a real-time system because the coefficients can vary over space and time. Few studies have been conducted to explore the optimal way to apply AOD data to improve model accuracies of PM2.5 surface estimation in a real-time air quality system. This paper discusses the best temporal scale to calculate the correlation of AOD and ground particle matter data to improve the results of pollution models in real-time system.

  20. AOD distributions and trends of major aerosol species over a selection of the world’s most populated cities based on the 1st Version of NASA’s MERRA Aerosol Reanalysis

    PubMed Central

    Provençal, Simon; Kishcha, Pavel; da Silva, Arlindo M.; Elhacham, Emily; Alpert, Pinhas

    2018-01-01

    NASA recently extended the Modern-Era Retrospective Analysis for Research and Application (MERRA) with an atmospheric aerosol reanalysis which includes five particulate species: sulfate, organic matter, black carbon, mineral dust and sea salt. The MERRA Aerosol Reanalysis (MERRAero) is an innovative tool to study air quality issues around the world for its global and constant coverage and its distinction of aerosol speciation expressed in the form of aerosol optical depth (AOD). The purpose of this manuscript is to apply MERRAero to the study of urban air pollution at the global scale by analyzing the AOD over a period of 13 years (2003–2015) and over a selection of 200 of the world’s most populated cities in order to assess the impacts of urbanization, industrialization, air quality regulations and regional transport which affect urban aerosol load. Environmental regulations and the recent global economic recession have helped to decrease the AOD and sulfate aerosols in most cities in North America, Europe and Japan. Rapid industrialization in China over the last two decades resulted in Chinese cities having the highest AOD values in the world. China has nevertheless recently implemented emission control measures which are showing early signs of success in many cities of Southern China where AOD has decreased substantially over the last 13 years. The AOD over South American cities, which is dominated by carbonaceous aerosols, has also decreased over the last decade due to an increase in commodity prices which slowed deforestation activities in the Amazon rainforest. At the opposite, recent urbanization and industrialization in India and Bangladesh resulted in a strong increase of AOD, sulfate and carbonaceous aerosols in most cities of these two countries. The AOD over most cities in Northern Africa and Western Asia changed little over the last decade. Emissions of natural aerosols, which cities in these two regions tend to be mostly composed of, don’t tend

  1. Troubled families and individualised solutions: an institutional discourse analysis of alcohol and drug treatment practices involving affected others.

    PubMed

    Selbekk, Anne Schanche; Sagvaag, Hildegunn

    2016-09-01

    Research shows that members of the families with patients suffering from alcohol and other drug-related issues (AOD) experience stress and strain. An important question is, what options do AOD treatment have for them when it comes to support? To answer this, we interviewed directors and clinicians from three AOD treatment institutions in Norway. The study revealed that family-oriented practices are gaining ground as a 'going concern'. However, the relative position of family-orientation in the services, is constrained and shaped by three other going concerns related to: (i) discourse on health and illness, emphasising that addiction is an individual medical and psychological phenomenon, rather than a relational one; (ii) discourse on rights and involvement, emphasising the autonomy of the individual patient and their right to define the format of their own treatment; and (iii) discourse on management, emphasising the relationship between cost and benefit, where family-oriented practices are defined as not being cost-effective. All three discourses are connected to underpin the weight placed on individualised practices. Thus, the findings point to a paradox: there is a growing focus on the needs of children and affected family members, while the possibility of performing integrated work on families is limited. © 2016 Foundation for the Sociology of Health & Illness.

  2. A Randomized Controlled Trial of a Group Motivational Interviewing Intervention for Adolescents with a First Time Alcohol or Drug Offense

    PubMed Central

    D’Amico, Elizabeth J.; Hunter, Sarah B.; Miles, Jeremy N.V.; Ewing, Brett A.; Osilla, Karen Chan

    2013-01-01

    Group Motivational Interviewing (MI) interventions that target youth at-risk for alcohol and other drug (AOD) use may prevent future negative consequences. Youth in a teen court setting (n=193; 67% male, 45% Hispanic; mean age 16.6 (SD = 1.05) were randomized to receive either a group MI intervention, Free Talk, or usual care (UC). We examined client acceptance, intervention feasibility and conducted a preliminary outcome evaluation. Free Talk teens reported higher quality and satisfaction ratings, and MI integrity scores were higher for Free Talk groups. AOD use and delinquency decreased for both groups at three months, and 12-month recidivism rates were lower but not significantly different for the Free Talk group compared to UC. Results contribute to emerging literature on MI in a group setting. A longer term follow-up is warranted. PMID:23891459

  3. Factors that enable and hinder the implementation of projects in the alcohol and other drug field.

    PubMed

    MacLean, Sarah; Berends, Lynda; Hunter, Barbara; Roberts, Bridget; Mugavin, Janette

    2012-02-01

    Few studies systematically explore elements of successful project implementation across a range of alcohol and other drug (AOD) activities. This paper provides an evidence base to inform project implementation in the AOD field. We accessed records for 127 completed projects funded by the Alcohol, Education and Rehabilitation Foundation from 2002 to 2008. An adapted realist synthesis methodology enabled us to develop categories of enablers and barriers to successful project implementation, and to identify factors statistically associated with successful project implementation, defined as meeting all funding objectives. Thematic analysis of eight case study projects allowed detailed exploration of findings. Nine enabler and 10 barrier categories were identified. Those most frequently reported as both barriers and enablers concerned partnerships with external agencies and communities, staffing and project design. Achieving supportive relationships with partner agencies and communities, employing skilled staff and implementing consumer or participant input mechanisms were statistically associated with successful project implementation. The framework described here will support development of evidence-based project funding guidelines and project performance indicators. The study provides evidence that investing project hours and resources to develop robust relationships with project partners and communities, implementing mechanisms for consumer or participant input and attracting skilled staff are legitimate and important activities, not just in themselves but because they potentially influence achievement of project funding objectives. © 2012 The Authors. ANZJPH © 2012 Public Health Association of Australia.

  4. Construction of an Inexpensive Sun Photometer for Measuring Aerosols Optical Depth (AOD) and Comparison Between the Ground Based and Satellite Based Measurements

    NASA Astrophysics Data System (ADS)

    Mamun, M.; Mondol, P.

    2012-12-01

    Aerosols influence our weather and climate because they affect the amount of sunlight reaching Earth's surface. An important way of probing the atmosphere from the ground is to measure the effects of the atmosphere on sunlight transmitted through the atmosphere to Earth's surface. These indirect techniques provide information about the entire atmosphere above the observer, not just the atmosphere that can be sampled directly. In response to global issues of air quality and climate change, and to the need to improve the quality of science education, inexpensive atmosphere monitoring instruments have been developed. This paper describes a new kind of inexpensive two channels LED Sun Photometer for monitoring aerosols that provide much better long-term stability than instruments that use expensive interference filters. Here HAZE-SPAN TERC VHS-1 model has been used for constructing sun photometer with light emitting diode as detector. Monitoring Earth's atmosphere is a challenging task. As there is no facility in our country (Bangladesh) for ground based measurement for monitoring aerosol so, this type of study is very essential. This study compares the aerosol optical depth (AOD) retrieved from the Terra and Aqua MODerate Resolution Imaging Spectroradiometers (MODIS) with ground-based measurements from a handheld sun photometer over the region of Rajshahi, Bangladesh for The 15 days duration of June 2012. The results indicate that the Terra and Aqua MODIS AOD retrievals at 550 nm have good correlations with the measurements from the handheld sun photometer. The correlation coefficients r = 0.88 for Terra and r = 0.55 for Aqua where as r = 0.65 for Terra and Aqua themselves. AOD for another wavelength at 625 nm is documented in this study for finding out the relation of AOD at different wavelengths. In this paper it has been described and summarized briefly investigations for four important topics: LEDs used as light detectors, construction of sun photometer and its

  5. NADPH oxidase inhibitors: a patent review.

    PubMed

    Kim, Jung-Ae; Neupane, Ganesh Prasad; Lee, Eung Seok; Jeong, Byeong-Seon; Park, Byung Chul; Thapa, Pritam

    2011-08-01

    NADPH oxidases, a family of multi-subunit enzyme complexes, catalyze the production of reactive oxygen species (ROS), which may contribute to the pathogenesis of a variety of diseases. In addition to the first NADPH oxidase found in phagocytes, four non-phagocytic NADPH oxidase isoforms have been identified, which all differ in their catalytic subunit (Nox1-5) and tissue distribution. This paper provides a comprehensive review of the patent literature on NADPH oxidase inhibitors, small molecule Nox inhibitors, peptides and siRNAs. Since each member of the NADPH oxidase family has great potential as a therapeutic target, several different compounds have been registered as NADPH oxidase inhibitors in the patent literature. As yet, none have gone through clinical trials, and some have not completed preclinical trials, including safety and specificity evaluation. Recently, small molecule pyrazolopyridine and triazolopyrimidine derivatives have been submitted as potent NADPH oxidase inhibitors and reported as first-in-class inhibitors for idiopathic pulmonary fibrosis and acute stroke, respectively. Further clinical efficacy and safety data are warranted to prove their actual clinical utility.

  6. A pilot test of a motivational interviewing social network intervention to reduce substance use among housing first residents.

    PubMed

    Kennedy, David P; Osilla, Karen Chan; Hunter, Sarah B; Golinelli, Daniela; Maksabedian Hernandez, Ervant; Tucker, Joan S

    2018-03-01

    This article presents findings of a pilot test of a Motivational Interviewing social network intervention (MI-SNI) to enhance motivation to reduce high risk alcohol and other drug (AOD) use among formerly homeless individuals transitioning to housing. Delivered in-person by a facilitator trained in MI, this four-session computer-assisted intervention provides personalized social network visualization feedback to help participants understand the people in their network who trigger their alcohol and other drug (AOD) use and those who support abstinence. If ready, participants are encouraged to make changes to their social network to help reduce their own high-risk behavior. Participants were 41 individuals (33 male, 7 female, 1 other; 23 African-American, 5 non-Latino White, 6 Latino, 7 other, mean age 48) who were transitioning from homelessness to permanent supportive housing. They were randomly assigned to either the MI-SNI condition or usual care. Readiness to change AOD use, AOD abstinence self-efficacy, and AOD use were assessed at baseline and shortly after the final intervention session for the MI-SNI arm and around 3-months after baseline for the control arm. Acceptability of the intervention was also evaluated. MI-SNI participants reported increased readiness to change AOD use compared to control participants. We also conducted a subsample analysis for participants at one housing program and found a significant intervention effect on readiness to change AOD use, AOD abstinence self-efficacy, and alcohol use compared to control participants. Participants rated the intervention as highly acceptable. We conclude that a brief computer-assisted Motivational Interviewing social network intervention has potential to efficaciously impact readiness to change AOD use, AOD abstinence self-efficacy, and AOD use among formerly homeless individuals transitioning to permanent supportive housing, and warrants future study in larger clinical trials. Copyright © 2017

  7. Estimation of surface PM10 concentration in Seoul during the DRAGON-Asia campaign based on the physical relationship between AOD and PM

    NASA Astrophysics Data System (ADS)

    Seo, S.; Kim, J.; Lee, H.; Jeong, U.; Kim, W. V.; Holben, B. N.; Kim, S.

    2013-12-01

    Atmospheric aerosols are known to play a role in climate change while it also adverse effects on human health such as respiratory and cardiovascular diseases. Especially, in terms of air quality, many studies have been conducted to estimate surface-level particulate matter (PM) concentration by using the satellite measurements to overcome the spatial limitation of ground-based aerosol measurements. In this study, we investigate the relationship between the column aerosol optical depth (AOD) and the surface PM10 concentration using the aerosol measurements during the DRAGON (Distributed Regional Aerosol Gridded Observation Network) - Asia campaign took place in Seoul from March to May, 2012. Based on the physical relationship between AOD and PM concentration, we develop various empirical linear models and evaluate the performance of these models. The best correlation (r = 0.67) is shown when vertical and size distribution of aerosols are additionally considered by using the boundary layer height (BLH) from backscattered lidar signals and the effective radius provided in AERONET inversion products. Similarly, MODIS AOD divided by BLH shows the best correlation with hourly PM10 (r = 0.62). We also identify the variability of correlations between AOD and PM10 depending on the environment characteristics in a complex megacity, Seoul by using the aerosol optical properties measured at mesoscale-level at 10 AERONET sites during the DRAGON campaign. Both AERONET and MODIS show higher correlation in residential area than near source area. Finally, we investigate the seasonal effects on the performance of various empirical linear models and find important factors of each season in PM estimation.

  8. Mit1 Transcription Factor Mediates Methanol Signaling and Regulates the Alcohol Oxidase 1 (AOX1) Promoter in Pichia pastoris.

    PubMed

    Wang, Xiaolong; Wang, Qi; Wang, Jinjia; Bai, Peng; Shi, Lei; Shen, Wei; Zhou, Mian; Zhou, Xiangshan; Zhang, Yuanxing; Cai, Menghao

    2016-03-18

    The alcohol oxidase 1 (AOX1) promoter (P AOX1) of Pichia pastoris is the most powerful and commonly used promoter for driving protein expression. However, mechanisms regulating its transcriptional activity are unclear. Here, we identified a Zn(II)2Cys6-type methanol-induced transcription factor 1 (Mit1) and elucidated its roles in regulating PAOX1 activity in response to glycerol and methanol. Mit1 regulated the expression of many genes involved in methanol utilization pathway, including AOX1, but did not participate in peroxisome proliferation and transportation of peroxisomal proteins during methanol metabolism. Structural analysis of Mit1 by performing domain deletions confirmed its specific and critical role in the strict repression of P AOX1 in glycerol medium. Importantly, Mit1, Mxr1, and Prm1, which positively regulated P AOX1 in response to methanol, were bound to P AOX1 at different sites and did not interact with each other. However, these factors cooperatively activated P AOX1 through a cascade. Mxr1 mainly functioned during carbon derepression, whereas Mit1 and Prm1 functioned during methanol induction, with Prm1 transmitting methanol signal to Mit1 by binding to the MIT1 promoter (P MIT1), thus increasingly expressing Mit1 and subsequently activating P AOX1. © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

  9. Tobacco smoking policies in Australian alcohol and other drug treatment services, agreement between staff awareness and the written policy document.

    PubMed

    Skelton, Eliza; Bonevski, Billie; Tzelepis, Flora; Shakeshaft, Anthony; Guillaumier, Ashleigh; Dunlop, Adrian; McCrabb, Sam; Palazzi, Kerrin

    2017-01-17

    Comprehensive smoke-free policy in the alcohol and other drug (AOD) setting provides an opportunity to reduce tobacco related harms among clients and staff. This study aimed to examine within AOD services: staff awareness of their service's smoking policy compared to the written policy document and staff and service factors associated with accurate awareness of a total ban and perceived enforcement of a total ban. An audit of written tobacco smoking policy documents and an online cross-sectional survey of staff from 31 Australian AOD services. In addition, a contact at each service was interviewed to gather service-related data. Overall, 506 staff participated in the survey (response rate: 57%). Nearly half (46%) perceived their service had a total ban with 54% indicating that this policy was always enforced. Over one-third (37%) reported a partial ban with 48% indicating that this policy was always enforced. The audit of written policies revealed that 19 (61%) services had total bans, 11 (36%) had partial bans and 1 (3%) did not have a written smoking policy. Agreement between staff policy awareness and their service's written policy was moderate (Kappa 0.48) for a total ban and fair (Kappa 0.38) for a partial ban. Age (1 year increase) of staff was associated with higher odds of correctly identifying a total ban at their service. Tobacco smoking within Australian AOD services is mostly regulated by a written policy document. Staff policy awareness was modest and perceived policy enforcement was poor.

  10. Validation of Satellite AOD Data with the Ground PM10 Data over Islamabad Pakistan

    NASA Astrophysics Data System (ADS)

    Bulbul, Gufran; Shahid, Imran

    2016-07-01

    health. In this study, concentrations of PM10 will be monitored at different sites in H-12 sector and Kashmir Highway Islamabad using High volume air sampler and its chemical characterization will be done using Energy Dispersive XRF. The first application of satellite remote sensing for aerosol monitoring began in the mid-1970s to detect the desert particles above the ocean using data from Landsat, GOES, and AVHRR remote sensing satellites. Maps of Aerosol Optical Depth (AOD) over the ocean were produced using the 0.63 µm channel of Advanced Very High Resolution Radiometer (AVHRR) . Aerosols properties were retrieved using AVHRR. The useable range of wavelengths of spectrum (shorter wavelengths and the longer wavelengths) for the remote sensing of the aerosols particles is mostly restricted due to ozone and gaseous absorptions. The purpose of the study is to validate the satellite Aerosol Optical Depth (AOD) data for the regional and local scale for Pakistan Objectives • To quantify the concentration of PM10 • To investigate their elemental composition • To find out their possible sources • Validation with MODIS satellite AOD Methodology: PM10 concentration will be measured at different sites of NUST Islamabad, Pakistan using High volume air sampler an Air sampling equipment capable of sampling high volumes of air (typically 57,000 ft3 or 1,600 m3) at high flow rates (typically 1.13 m3/min or 40 ft3/min) over an extended sampling duration (typically 24 hrs). The sampling period will be of 24 hours. Particles in the PM10 size range are then collected on the filter(s) during the specified 24-h sampling period. Each sample filter will be weighed before and after sampling to determine the net weight (mass) gain of the collected PM10 sample (40 CFR Part 50, Appendix M, US EPA). Next step will be the chemical characterization. Element concentrations will be determined by energy dispersive X-ray fluorescence (ED-XRF) technique. The ED-XRF system uses an X-ray tube to

  11. Drug and alcohol-impaired driving among electronic music dance event attendees

    PubMed Central

    Furr-Holden, Debra; Voas, Robert B.; Kelley-Baker, Tara; Miller, Brenda

    2011-01-01

    Background Drug-impaired driving has received increased attention resulting from development of rapid drug-screening procedures used by police and state laws establishing per se limits for drug levels in drivers. Venues that host electronic music dance events (EMDEs) provide a unique opportunity to assess drug-impaired driving among a high proportion of young adult drug users. EMDEs are late-night dance parties marked by a substantial number of young adult attendees and elevated drug involvement. No studies to date have examined drug-impaired driving in a natural environment with active drug and alcohol users. Methods Six EMDEs were sampled in San Diego, California, and Baltimore, Maryland. A random sample of approximately 40 attendees per event were administered surveys about alcohol and other drug (AOD) use and driving status, given breath tests for alcohol, and asked to provide oral fluid samples to test for illicit drug use upon entering and exiting the events. Results Driving status reduced the level of alcohol use (including abstaining) but the impact on drug-taking was not significant. However, 62% of individuals who reported their intention to drive away from the events were positive for drugs or alcohol upon leaving. This suggests that these events and settings are appropriate ones for developing interventions for reducing risks for young adults. PMID:16675160

  12. Targeting NADPH oxidases in vascular pharmacology

    PubMed Central

    Schramm, Agata; Matusik, Paweł; Osmenda, Grzegorz; Guzik, Tomasz J

    2012-01-01

    Oxidative stress is a molecular dysregulation in reactive oxygen species (ROS) metabolism, which plays a key role in the pathogenesis of atherosclerosis, vascular inflammation and endothelial dysfunction. It is characterized by a loss of nitric oxide (NO) bioavailability. Large clinical trials such as HOPE and HPS have not shown a clinical benefit of antioxidant vitamin C or vitamin E treatment, putting into question the role of oxidative stress in cardiovascular disease. A change in the understanding of the molecular nature of oxidative stress has been driven by the results of these trials. Oxidative stress is no longer perceived as a simple imbalance between the production and scavenging of ROS, but as a dysfunction of enzymes involved in ROS production. NADPH oxidases are at the center of these events, underlying the dysfunction of other oxidases including eNOS uncoupling, xanthine oxidase and mitochondrial dysfunction. Thus NADPH oxidases are important therapeutic targets. Indeed, HMG-CoA reductase inhibitors (statins) as well as drugs interfering with the renin-angiotensin-aldosterone system inhibit NADPH oxidase activation and expression. Angiotensin-converting enzyme (ACE) inhibitors, AT1 receptor antagonists (sartans) and aliskiren, as well as spironolactone or eplerenone, have been discussed. Molecular aspects of NADPH oxidase regulation must be considered, while thinking about novel pharmacological targeting of this family of enzymes consisting of several homologs Nox1, Nox2, Nox3, Nox4 and Nox5 in humans. In order to properly design trials of antioxidant therapies, we must develop reliable techniques for the assessment of local and systemic oxidative stress. Classical antioxidants could be combined with novel oxidase inhibitors. In this review, we discuss NADPH oxidase inhibitors such as VAS2870, VAS3947, GK-136901, S17834 or plumbagin. Therefore, our efforts must focus on generating small molecular weight inhibitors of NADPH oxidases, allowing the

  13. Substance Use Attitudes and Behaviors at Three Pharmacy Colleges

    ERIC Educational Resources Information Center

    Baldwin, Jeffrey N.; Scott, David M.; DeSimone, Edward M., II; Forrester, Joy H.; Fankhauser, Martha P.

    2011-01-01

    The objective of this study was to profile and compare alcohol and other drug (AOD) use attitudes and behaviors in three pharmacy colleges. Student surveys of AOD use attitudes and behaviors were conducted at one southwestern and two midwestern pharmacy colleges. Response was 86.5% (566/654). Reported past-year use included alcohol 82.8%, tobacco…

  14. MAOA expression predicts vulnerability for alcohol use.

    PubMed

    Cervera-Juanes, R; Wilhem, L J; Park, B; Lee, R; Locke, J; Helms, C; Gonzales, S; Wand, G; Jones, S R; Grant, K A; Ferguson, B

    2016-04-01

    The role of the monoamines dopamine (DA) and serotonin (5HT) and the monoamine-metabolizing enzyme monoamine oxidase A (MAOA) have been repeatedly implicated in studies of alcohol use and dependence. Genetic investigations of MAOA have yielded conflicting associations between a common polymorphism (MAOA-LPR) and risk for alcohol abuse. The present study provides direct comparison of tissue-specific MAOA expression and the level of alcohol consumption. We analyzed rhesus macaque MAOA (rhMAOA) expression in blood from males before and after 12 months of alcohol self-administration. In addition, nucleus accumbens core (NAc core) and cerebrospinal fluid (CSF) were collected from alcohol access and control (no alcohol access) subjects at the 12-month time point for comparison. The rhMAOA expression level in the blood of alcohol-naive subjects was negatively correlated with subsequent alcohol consumption level. The mRNA expression was independent of rhMAOA-LPR genotype and global promoter methylation. After 12 months of alcohol use, blood rhMAOA expression had decreased in an alcohol dose-dependent manner. Also after 12 months, rhMAOA expression in the NAc core was significantly lower in the heavy drinkers, as compared with control subjects. The CSF measured higher levels of DA and lower DOPAC/DA ratios among the heavy drinkers at the same time point. These results provide novel evidence that blood MAOA expression predicts alcohol consumption and that heavy alcohol use is linked to low MAOA expression in both the blood and NAc core. Together, the findings suggest a mechanistic link between dampened MAOA expression, elevated DA and alcohol abuse.

  15. Technical Report Series on Global Modeling and Data Assimilation. Volume 32; Estimates of AOD Trends (2002 - 2012) Over the World's Major Cities Based on the MERRA Aerosol Reanalysis

    NASA Technical Reports Server (NTRS)

    Provencal, Simon; Kishcha, Pavel; Elhacham, Emily; daSilva, Arlindo M.; Alpert, Pinhas; Suarez, Max J.

    2014-01-01

    NASA's Global Modeling and Assimilation Office has extended the Modern-Era Retrospective Analysis for Research and Application (MERRA) tool with five atmospheric aerosol species (sulfates, organic carbon, black carbon, mineral dust and sea salt). This inclusion of aerosol reanalysis data is now known as MERRAero. This study analyses a ten-year period (July 2002 - June 2012) MERRAero aerosol reanalysis applied to the study of aerosol optical depth (AOD) and its trends for the aforementioned aerosol species over the world's major cities (with a population of over 2 million inhabitants). We found that a proportion of various aerosol species in total AOD exhibited a geographical dependence. Cities in industrialized regions (North America, Europe, central and eastern Asia) are characterized by a strong proportion of sulfate aerosols. Organic carbon aerosols are dominant over cities which are located in regions where biomass burning frequently occurs (South America and southern Africa). Mineral dust dominates other aerosol species in cities located in proximity to the major deserts (northern Africa and western Asia). Sea salt aerosols are prominent in coastal cities but are dominant aerosol species in very few of them. AOD trends are declining over cities in North America, Europe and Japan, as a result of effective air quality regulation. By contrast, the economic boom in China and India has led to increasing AOD trends over most cities in these two highly-populated countries. Increasing AOD trends over cities in the Middle East are caused by increasing desert dust.

  16. Using optimal interpolation to assimilate surface measurements and satellite AOD for ozone and PM2.5: A case study for July 2011.

    PubMed

    Tang, Youhua; Chai, Tianfeng; Pan, Li; Lee, Pius; Tong, Daniel; Kim, Hyun-Cheol; Chen, Weiwei

    2015-10-01

    We employed an optimal interpolation (OI) method to assimilate AIRNow ozone/PM2.5 and MODIS (Moderate Resolution Imaging Spectroradiometer) aerosol optical depth (AOD) data into the Community Multi-scale Air Quality (CMAQ) model to improve the ozone and total aerosol concentration for the CMAQ simulation over the contiguous United States (CONUS). AIRNow data assimilation was applied to the boundary layer, and MODIS AOD data were used to adjust total column aerosol. Four OI cases were designed to examine the effects of uncertainty setting and assimilation time; two of these cases used uncertainties that varied in time and location, or "dynamic uncertainties." More frequent assimilation and higher model uncertainties pushed the modeled results closer to the observation. Our comparison over a 24-hr period showed that ozone and PM2.5 mean biases could be reduced from 2.54 ppbV to 1.06 ppbV and from -7.14 µg/m³ to -0.11 µg/m³, respectively, over CONUS, while their correlations were also improved. Comparison to DISCOVER-AQ 2011 aircraft measurement showed that surface ozone assimilation applied to the CMAQ simulation improves regional low-altitude (below 2 km) ozone simulation. This paper described an application of using optimal interpolation method to improve the model's ozone and PM2.5 estimation using surface measurement and satellite AOD. It highlights the usage of the operational AIRNow data set, which is available in near real time, and the MODIS AOD. With a similar method, we can also use other satellite products, such as the latest VIIRS products, to improve PM2.5 prediction.

  17. Isolated sulfite oxidase deficiency.

    PubMed

    Rupar, C A; Gillett, J; Gordon, B A; Ramsay, D A; Johnson, J L; Garrett, R M; Rajagopalan, K V; Jung, J H; Bacheyie, G S; Sellers, A R

    1996-12-01

    Isolated sulfite oxidase (SO) deficiency is an autosomal recessively inherited inborn error of sulfur metabolism. In this report of a ninth patient the clinical history, laboratory results, neuropathological findings and a mutation in the sulfite oxidase gene are described. The data from this patient and previously published patients with isolated sulfite oxidase deficiency and molybdenum cofactor deficiency are summarized to characterize this rare disorder. The patient presented neonatally with intractable seizures and did not progress developmentally beyond the neonatal stage. Dislocated lenses were apparent at 2 months. There was increased urine excretion of sulfite and S-sulfocysteine and a decreased concentration of plasma cystine. A lactic acidemia was present for 6 months. Liver sulfite oxidase activity was not detectable but xanthine dehydrogenase activity was normal. The boy died of respiratory failure at 32 months. Neuropathological findings of cortical necrosis and extensive cavitating leukoencephalopathy were reminiscent of those seen in severe perinatal asphyxia suggesting an etiology of energy deficiency. A point mutation that resulted in a truncated protein missing the molybdenum-binding site has been identified.

  18. Continuous minimally-invasive alcohol monitoring using microneedle sensor arrays

    PubMed Central

    Vinu Mohan, A. M.; Windmiller, Joshua Ray; Mishra, Rupesh K.; Wang, Joseph

    2017-01-01

    The present work describes an attractive skin-worn microneedle sensing device for the minimally invasive electrochemical monitoring of subcutaneous alcohol. The device consists of an assembly of pyramidal microneedle structures integrated with Pt and Ag wires, each with a microcavity opening. The microneedle aperture was modified by electropolymerizing o-phenylene diamine onto the Pt wire microtransducer, followed by the immobilization of alcohol oxidase (AOx) in an intermediate chitosan layer, along with an outer Nafion layer. The resulting microneedle-based enzyme electrode displays an interference-free ethanol detection in artificial interstitial fluid without compromising its sensitivity, stability and response time. The skin penetration ability and the efficaciousness of the biosensor performance towards subcutaneous alcohol monitoring was substantiated by the ex vivo mice skin model analysis. Our results reveal that the new microneedle sensor holds considerable promise for continuous non-invasive alcohol monitoring in real-life situations. PMID:28088750

  19. NADPH Oxidases in Vascular Pathology

    PubMed Central

    Konior, Anna; Schramm, Agata; Czesnikiewicz-Guzik, Marta

    2014-01-01

    Abstract Significance: Reactive oxygen species (ROS) play a critical role in vascular disease. While there are many possible sources of ROS, nicotinamide adenine dinucleotide phosphate (NADPH) oxidases play a central role. They are a source of “kindling radicals,” which affect other enzymes, such as nitric oxide synthase endothelial nitric oxide synthase or xanthine oxidase. This is important, as risk factors for atherosclerosis (hypertension, diabetes, hypercholesterolemia, and smoking) regulate the expression and activity of NADPH oxidases in the vessel wall. Recent Advances: There are seven isoforms in mammals: Nox1, Nox2, Nox3, Nox4, Nox5, Duox1 and Duox2. Nox1, Nox2, Nox4, and Nox5 are expressed in endothelium, vascular smooth muscle cells, fibroblasts, or perivascular adipocytes. Other homologues have not been found or are expressed at very low levels; their roles have not been established. Nox1/Nox2 promote the development of endothelial dysfunction, hypertension, and inflammation. Nox4 may have a role in protecting the vasculature during stress; however, when its activity is increased, it may be detrimental. Calcium-dependent Nox5 has been implicated in oxidative damage in human atherosclerosis. Critical Issues: NADPH oxidase-derived ROS play a role in vascular pathology as well as in the maintenance of normal physiological vascular function. We also discuss recently elucidated mechanisms such as the role of NADPH oxidases in vascular protection, vascular inflammation, pulmonary hypertension, tumor angiogenesis, and central nervous system regulation of vascular function and hypertension. Future Directions: Understanding the role of individual oxidases and interactions between homologues in vascular disease is critical for efficient pharmacological regulation of vascular NADPH oxidases in both the laboratory and clinical practice. Antioxid. Redox Signal. 20, 2794–2814. PMID:24180474

  20. Oxidase positive rods from cases of suspected gonorrhoea. A comparison of conventional, gas chromatographic and genetic methods of identification.

    PubMed

    Bovre, K; Hagen, N; Berdal, B P; Jantzen, E

    1977-02-01

    Genito-urethral specimens from 3260 women and 1170 men, with ailments suggestive of gonorrhoea, were examined for growth of oxidase positive rodshaped bacteria, as well as of gonococci. Moraxella osloensis was identified in 26 cases (0.64 per cent of women and 0.43 per cent of men). Three patients harboured phenylalanine negative (or weakly reacting) and tryptophan deaminase negative M. phenylpyrouvica and, in three cases, a Flavobacterium species was detected. Among six oropharyngeal specimens from patients suspected of gonorrhoea, two yielded growth of oxidase positive rods, Kingella kingae and Neisseria elongata, respectively, N. gonorrhoeae was isolated from 537 patients, i.e., 12.1 per cent of all cases. The isolates of oxidase positive rods were in most cases completely identified by streptomycin resistance transformation. On this basis, the diagnostic reliability of some morphological and cultural-biochemical tests and gas chromatography was examined. Gas chromatographic analysis of fatty acid and alcohol composition of whole cells proved distinctive of species defined genetically, irrespective of confusing behaviour of some strains in other tests.

  1. Continuous minimally-invasive alcohol monitoring using microneedle sensor arrays.

    PubMed

    Mohan, A M Vinu; Windmiller, Joshua Ray; Mishra, Rupesh K; Wang, Joseph

    2017-05-15

    The present work describes an attractive skin-worn microneedle sensing device for the minimally invasive electrochemical monitoring of subcutaneous alcohol. The device consists of an assembly of pyramidal microneedle structures integrated with Pt and Ag wires, each with a microcavity opening. The microneedle aperture was modified by electropolymerizing o-phenylene diamine onto the Pt wire microtransducer, followed by the immobilization of alcohol oxidase (AOx) in an intermediate chitosan layer, along with an outer Nafion layer. The resulting microneedle-based enzyme electrode displays an interference-free ethanol detection in artificial interstitial fluid without compromising its sensitivity, stability and response time. The skin penetration ability and the efficaciousness of the biosensor performance towards subcutaneous alcohol monitoring was substantiated by the ex vivo mice skin model analysis. Our results reveal that the new microneedle sensor holds considerable promise for continuous non-invasive alcohol monitoring in real-life situations. Copyright © 2017 Elsevier B.V. All rights reserved.

  2. SPERMINE OXIDASE: AN AMINE OXIDASE WITH SPECIFICITY FOR SPERMINE AND SPERMIDINE

    PubMed Central

    Hirsch, James G.

    1953-01-01

    Sheep serum and bovine serum contain an enzyme which brings about a rapid oxidative deamination of certain biological amines. This enzyme differs from previously described amine oxidases in several regards and especially in its substrate specificity. Studies thus far indicate that only spermine and the closely related compound spermidine serve as substrates for the enzyme in sheep serum. For this reason, the enzyme has been named spermine oxidase. Spermine oxidase is active in a variety of fluids of various ionic strength and buffer composition. The reaction takes place between pH 6.0 and pH 8.0 with an optimal rate in the vicinity of neutrality. Under certain conditions, the rate of oxygen consumption during the initial phase of the reaction is independent of the concentration of substrate. The diminution in rate observed during the latter phase of the enzymatic attack appears to be due to an alteration in the kinetics at low concentrations of substrate, or to competitive inhibition by a product of the reaction. Carbonyl reagents almost completely block the action of spermine oxidase, while certain amines and the cyanide ion bring about partial inhibition. Thiol reagents and sequestering compounds do not alter the course of the oxidative process. In the presence of low concentrations of mercuric chloride, the sheep serum-spermine system consumes approximately twice as much oxygen as controls containing no mercuric ion. The mechanism by which the mercuric ion stimulates additional oxygen uptake is obscure. PMID:13052805

  3. Early Intervention: How Parents Can Help Adolescent Children Who May Be Using Drugs.

    ERIC Educational Resources Information Center

    Franklin, Diane, Ed.; Bankston, Karen, Ed.

    The earlier a parent can detect and respond to a child's involvement with alcohol and other drugs (AOD), the better. Just as early detection heightens the cure rate for cancer, so too does early intervention increase the chances of ending AOD use. This booklet outlines warning signs of AOD use and lists the hazards of adolescent drug use.…

  4. Analysis of the 5′ untranslated region (5′UTR) of the alcohol oxidase 1 (AOX1) gene in recombinant protein expression in Pichia pastoris

    PubMed Central

    Staley, Chris A.; Huang, Amy; Nattestad, Maria; Oshiro, Kristin T.; Ray, Laura E.; Mulye, Tejas; Li, Zhiguo Harry; Le, Thu; Stephens, Justin J.; Gomez, Seth R.; Moy, Allison D.; Nguyen, Jackson C.; Franz, Andreas H.; Lin-Cereghino, Joan; Lin-Cereghino, Geoff P.

    2012-01-01

    Pichia pastoris is a methylotrophic yeast that has been genetically engineered to express over one thousand heterologous proteins valued for industrial, pharmaceutical and basic research purposes. In most cases, the 5′ untranslated region (UTR) of the alcohol oxidase 1 (AOX1) gene is fused to the coding sequence of the recombinant gene for protein expression in this yeast. Because the effect of the AOX1 5′UTR on protein expression is not known, site-directed mutagenesis was performed in order to decrease or increase the length of this region. Both of these types of changes were shown to affect translational efficiency, not transcript stability. While increasing the length of the 5′UTR clearly decreased expression of a β-galactosidase reporter in a proportional manner, a deletion analysis demonstrated that the AOX1 5′UTR contains a complex mixture of both positive and negative cis-acting elements, suggesting that the construction of a synthetic 5′UTR optimized for a higher level of expression may be challenging. PMID:22285974

  5. Cultural Identity Among Urban American Indian/Alaska Native Youth: Implications for Alcohol and Drug Use.

    PubMed

    Brown, Ryan A; Dickerson, Daniel L; D'Amico, Elizabeth J

    2016-10-01

    American Indian / Alaska Native (AI/AN) youth exhibit high rates of alcohol and other drug (AOD) use, which is often linked to the social and cultural upheaval experienced by AI/ANs during the colonization of North America. Urban AI/AN youth may face unique challenges, including increased acculturative stress due to lower concentrations of AI/AN populations in urban areas. Few existing studies have explored cultural identity among urban AI/AN youth and its association with AOD use. This study used systematic qualitative methods with AI/AN communities in two urban areas within California to shed light on how urban AI/AN youth construct cultural identity and how this relates to AOD use and risk behaviors. We conducted 10 focus groups with a total of 70 youth, parents, providers, and Community Advisory Board members and used team-based structured thematic analysis in the Dedoose software platform. We identified 12 themes: intergenerational stressors, cultural disconnection, AI/AN identity as protective, pan-tribal identity, mixed racial-ethnic identity, rural vs. urban environments, the importance of AI/AN institutions, stereotypes and harassment, cultural pride, developmental trajectories, risks of being AI/AN, and mainstream culture clash. Overall, youth voiced curiosity about their AI/AN roots and expressed interest in deepening their involvement in cultural activities. Adults described the myriad ways in which involvement in cultural activities provides therapeutic benefits for AI/AN youth. Interventions that provide urban AI/AN youth with an opportunity to engage in cultural activities and connect with positive and healthy constructs in AI/AN culture may provide added impact to existing interventions.

  6. Evaluation of Different MODIS AOD Retrieval Algorithms for PM2.5 Estimation in the Western, Midwestern and Southeastern United States with Implications for Public Health

    NASA Astrophysics Data System (ADS)

    Al-Hamdan, M. Z.; Crosson, W. L.; Burrows, E. C.; Coffield, S.; Crane, B.

    2016-12-01

    This study was part of the research activities of the Center for Applied Atmospheric Research and Education (CAARE) funded by the NASA MUREP Institutional Research Opportunity (MIRO) Program. Satellite measurements of Aerosol Optical Depth (AOD) have been shown to be correlated with ground measurements of fine particulate matter less than 2.5 microns (PM2.5), which in turn has been linked to respiratory and heart diseases. The strength of the correlation between AOD and PM2.5 varies for different AOD retrieval algorithms and geographic regions. We evaluated several Moderate Resolution Imaging Spectrometer (MODIS) AOD products from different satellites (Aqua vs. Terra), retrieval algorithms (Dark Target vs. Deep Blue), Collections (5.1 vs. 6) and spatial resolutions (10-km vs. 3-km) for cities in the Western, Midwestern and Southeastern United States. We developed and validated PM2.5 prediction models using remotely sensed AOD data, which were improved by incorporating meteorological variables (temperature, relative humidity, precipitation, wind speed, and wind direction) from the North American Land Data Assimilation System Phase 2 (NLDAS-2). Adding these meteorological data significantly improved the predictive power of all the PM2.5 models, and especially in the Western U.S. Temperature, relative humidity and wind speed were the most significant meteorological variables throughout the year in the Western U.S. Wind speed was the most significant meteorological variable for the cold season while temperature was the most significant variable for the warm season in the Midwestern and Southeastern U.S. Finally, our study re-establishes the connection between PM2.5 and public health concerns including respiratory and cardiovascular diseases (asthma, high blood pressure, coronary heart disease, heart attack, and stroke). Using PM2.5 data and health data from the Centers for Disease Control and Prevention (CDC)'s Behavioral Risk Factor Surveillance System (BRFSS), our

  7. Adipogenesis-related increase of semicarbazide-sensitive amine oxidase and monoamine oxidase in human adipocytes.

    PubMed

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

    2007-08-01

    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.

  8. Various applications of immobilized glucose oxidase and polyphenol oxidase in a conducting polymer matrix.

    PubMed

    Cil, M; Böyükbayram, A E; Kiralp, S; Toppare, L; Yağci, Y

    2007-06-01

    In this study, glucose oxidase and polyphenol oxidase were immobilized in conducting polymer matrices; polypyrrole and poly(N-(4-(3-thienyl methylene)-oxycarbonyl phenyl) maleimide-co-pyrrole) via electrochemical method. Fourier transform infrared and scanning electron microscope were employed to characterize the copolymer of (N-(4-(3-thienyl methylene)-oxycarbonyl phenyl) maleimide) with pyrrole. Kinetic parameters, maximum reaction rate and Michealis-Menten constant, were determined. Effects of temperature and pH were examined for immobilized enzymes. Also, storage and operational stabilities of enzyme electrodes were investigated. Glucose and polyphenol oxidase enzyme electrodes were used for determination of the glucose amount in orange juices and human serum and phenolic amount in red wines, respectively.

  9. Screening for alcohol and drug use in pregnancy.

    PubMed

    Seib, Charrlotte A; Daglish, Mark; Heath, Renée; Booker, Catriona; Reid, Carol; Fraser, Jennifer

    2012-12-01

    this study examined the clinical utility and precision of routine screening for alcohol and other drug use among women attending a public antenatal service. a survey of clients and audit of clinical charts. clients attending an antenatal clinic of a large tertiary hospital in Queensland, Australia, from October to December 2009. data were collected from two sources. First, 32 women who reported use of alcohol or other drugs during pregnancy at initial screening were then asked to complete a full substance use survey. Second, data were collected from charts of 349 new clients who attended the antenatal clinic during the study period. Both sensitivity (86%, 67%) and positive predictive value (100%, 92%) for alcohol and other drug use respectively, were high. Only 15% of surveyed women were uncomfortable about being screened for substance use in pregnancy, yet the chart audit revealed poor staff compliance. During the study period, 25% of clients were either not screened adequately or not at all. KEY CONCLUSIONS AND IMPLICATIONS FOR PRACTISE: despite recommended universal screening in pregnancy and the apparent acceptance by our participants, alcohol and other drug (A&OD) screening in the antenatal setting remains problematic. Investigation into the reasons behind, and ways to overcome, the low screening rate could improve health outcomes for mothers and children in this at-risk group. Targeted education and training for midwives may form part of the solution as these clinicians have a key role in implementing prevention and early intervention strategies. Copyright © 2011 Elsevier Ltd. All rights reserved.

  10. Screening for atypical suicide risk with person fit statistics among people presenting to alcohol and other drug treatment.

    PubMed

    Conrad, Kendon J; Bezruczko, Nikolaus; Chan, Ya-Fen; Riley, Barth; Diamond, Guy; Dennis, Michael L

    2010-01-15

    Symptoms of internalizing disorders (depression, anxiety, somatic, trauma) are the major risk factors for suicide. Atypical suicide risk is characterized by people with few or no symptoms of internalizing disorders. In persons screened at intake to alcohol or other drug (AOD) treatment, this research examined whether person fit statistics would support an atypical subtype at high risk for suicide that did not present with typical depression and other internalizing disorders. Symptom profiles of the prototypical, typical, and atypical persons, as defined using fit statistics, were tested on 7408 persons entering AOD treatment using the Global Appraisal of Individual Needs (GAIN; Dennis et al., 2003a,b). Of those with suicide symptoms, the findings were as expected with the atypical group being higher on suicide and lower on symptoms of internalizing disorders. In addition, the atypical group was similar or lower on substance problems, symptoms of externalizing disorders, and crime and violence. Person fit statistics were useful in identifying persons with atypical suicide profiles and in enlightening aspects of existing theory concerning atypical suicidal ideation. Copyright (c) 2009 Elsevier Ireland Ltd. All rights reserved.

  11. Articulating addiction in alcohol and other drug policy: A multiverse of habits.

    PubMed

    Fraser, Suzanne

    2016-05-01

    Concepts of addiction differ across time and place. This article is based on an international research project currently exploring this variation and change in concepts of addiction, in particular in the field of alcohol and other drug (AOD) use. Taking AOD policy in Australia and Canada as its empirical focus, and in-depth interviews with policy makers, service providers and advocates in each country as its key method (N=60), the article compares the addiction concepts articulated by professionals working in each setting. Drawing on Bruno Latour's theoretical work on the body and his proposal for a better science based on the 'articulation of differences', it explores the accounts of addiction offered across the Australian and Canadian project sites, identifying a shared dynamic in all: the juggling of difference and unity in discussions of the nature of addiction, its composite parts and how best to respond to it. The article maps two simultaneous trajectories in the data - one moving towards difference in participants' insistence on the multitude and diversity of factors that make up addiction problems and solutions, and the other towards unity in their tendency to return to narrow disease models of addiction in uncomfortable, sometimes dissonant, strategic choices. As I will argue, the AOD professionals interviewed for my project operate in two modes treated as distinct in Latour's proposal: in turning to reifying disease labels of addiction they take for granted, and work within, a 'universe of essences', but in articulating the multiplicity and diversity of addiction, they grope towards a vision of a 'multiverse of habits'. The article concludes by addressing this tension directly, scrutinising its practical implications for the development of policy and delivery of services in the future, asking how new thinking, and therefore new opportunities, might be allowed to emerge. Crown Copyright © 2015. Published by Elsevier B.V. All rights reserved.

  12. Inferring wavelength dependence of AOD and Ångström exponent over a sub-tropical station in South Africa using AERONET data: influence of meteorology, long-range transport and curvature effect.

    PubMed

    Kumar, K Raghavendra; Sivakumar, V; Reddy, R R; Gopal, K Rama; Adesina, A Joseph

    2013-09-01

    Aerosol optical properties over a southern sub-tropical site Skukuza, South Africa were studied to determine the variability of the aerosol characteristics using CIMEL Sunphotometer data as part of the AErosol RObotic NETwork (AERONET) from December 2005 to November 2006. Aerosol optical depth (AOD), Ångström exponent (α), and columnar water vapor (CWV) data were collected, analyzed, and compiled. Participating in this network provided a unique opportunity for understanding the sources of aerosols affecting the atmosphere of South Africa (SA) and the regional radiation budget. The meteorological patterns significantly (p<0.05) influenced the amount and size distribution of the aerosols. Results showed that seasonal variation of AOD at 500 nm (AOD500) over the observation site were characterized by low values (0.10-0.13) in autumn, moderate values (0.14-0.16) in summer and winter seasons, and high to very high values (0.18-0.40) during the spring, with an overall mean value of 0.18±0.12. Ångström exponent α(440-870), varied from 0.5 to 2.89, with significant (p<0.0001) seasonal variability. CWV showed a strong annual cycle with maximum values in the summer and autumn seasons. The relationship between AOD, Ångström exponent (α), and CWV showed a strong dependence (p<0.0001) of α on AOD and CWV, while there was no significant correlation between AOD and CWV. Investigation of the adequacy of the simple use of the spectral AOD and Ångström exponent data was used in deriving the curvature (a2) showed to obtain information for determining the aerosol-particle size. The negative a2 values are characterized by aerosol-size dominated by fine-mode (0.1-1 μm), while the positive curvatures indicate abundance of coarse particles (>1 μm). Trajectory cluster analyses revealed that the air masses during the autumn and winter seasons have longer advection pathways, passing over the ocean and continent. This is reflected in the aerosol properties that are derived from

  13. An Oxidase-Based Electrochemical Fluidic Sensor with High-Sensitivity and Low-Interference by On-Chip Oxygen Manipulation

    PubMed Central

    Radhakrishnan, Nitin; Park, Jongwon; Kim, Chang-Soo

    2012-01-01

    Utilizing a simple fluidic structure, we demonstrate the improved performance of oxidase-based enzymatic biosensors. Electrolysis of water is utilized to generate bubbles to manipulate the oxygen microenvironment close to the biosensor in a fluidic channel. For the proper enzyme reactions to occur, a simple mechanical procedure of manipulating bubbles was developed to maximize the oxygen level while minimizing the pH change after electrolysis. The sensors show improved sensitivities based on the oxygen dependency of enzyme reaction. In addition, this oxygen-rich operation minimizes the ratio of electrochemical interference signal by ascorbic acid during sensor operation (i.e., amperometric detection of hydrogen peroxide). Although creatinine sensors have been used as the model system in this study, this method is applicable to many other biosensors that can use oxidase enzymes (e.g., glucose, alcohol, phenol, etc.) to implement a viable component for in-line fluidic sensor systems. PMID:23012527

  14. Isolation and characterization of mutated alcohol oxidases from the yeast Hansenula polymorpha with decreased affinity toward substrates and their use as selective elements of an amperometric biosensor

    PubMed Central

    Dmytruk, Kostyantyn V; Smutok, Oleh V; Ryabova, Olena B; Gayda, Galyna Z; Sibirny, Volodymyr A; Schuhmann, Wolfgang; Gonchar, Mykhailo V; Sibirny, Andriy A

    2007-01-01

    Background Accurate, rapid, and economic on-line analysis of ethanol is very desirable. However, available biosensors achieve saturation at very low ethanol concentrations and thus demand the time and labour consuming procedure of sample dilution. Results Hansenula polymorpha (Pichia angusta) mutant strains resistant to allyl alcohol in methanol medium were selected. Such strains possessed decreased affinity of alcohol oxidase (AOX) towards methanol: the KM values for AOX of wild type and mutant strains CA2 and CA4 are shown to be 0.62, 2.48 and 1.10 mM, respectively, whereas Vmax values are increased or remain unaffected. The mutant AOX alleles from H. polymorpha mutants CA2 and CA4 were isolated and sequenced. Several point mutations in the AOX gene, mostly different between the two mutant alleles, have been identified. Mutant AOX forms were isolated and purified, and some of their biochemical properties were studied. An amperometric biosensor based on the mutated form of AOX from the strain CA2 was constructed and revealed an extended linear response to the target analytes, ethanol and formaldehyde, as compared to the sensor based on the native AOX. Conclusion The described selection methodology opens up the possibility of isolating modified forms of AOX with further decreased affinity toward substrates without reduction of the maximal velocity of reaction. It can help in creation of improved ethanol biosensors with a prolonged linear response towards ethanol in real samples of wines, beers or fermentation liquids. PMID:17567895

  15. Evaluation of Different MODIS AOD Retrieval Algorithms for PM (sub 2.5) Estimation in the Western, Midwestern and Southeastern United States with Implications for Public Health

    NASA Technical Reports Server (NTRS)

    Al-Hamdan, Mohammad; Crosson, William; Burrows, Erica; Coffield, Shane; Crane, Breanna

    2016-01-01

    This study was part of the research activities of the Center for Applied Atmospheric Research and Education (CAARE) funded by the NASA MUREP (Minority University Research and Education Project) Institutional Research Opportunity (MIRO) Program. Satellite measurements of Aerosol Optical Depth (AOD) have been shown to be correlated with ground measurements of fine particulate matter less than 2.5 microns PM (sub 2.5), which in turn has been linked to respiratory and heart diseases. The strength of the correlation between AOD and PM (sub 2.5) varies for different AOD retrieval algorithms and geographic regions. We evaluated several Moderate Resolution Imaging Spectrometer (MODIS) AOD products from different satellites (Aqua vs. Terra), retrieval algorithms (Dark Target versus Deep Blue), Collections (5.1 versus 6) and spatial resolutions (10-kilometers versus 3-kilometers) for cities in the Western, Midwestern and Southeastern U.S. We developed and validated PM (sub 2.5) prediction models using remotely-sensed AOD data, which were improved by incorporating meteorological variables (temperature, relative humidity, precipitation, wind speed, and wind direction) from the North American Land Data Assimilation System Phase 2 (NLDAS-2). Adding these meteorological data significantly improved the predictive power of all the PM (sub 2.5) models, especially in the Western U.S. Temperature, relative humidity and wind speed were the most significant meteorological variables throughout the year in the Western U.S. Wind speed was the most significant meteorological variable for the cold season while temperature was the most significant variable for the warm season in the Midwestern and Southeastern U.S. Our study re-establishes the connection between PM (sub 2.5) and public health concerns including respiratory and cardiovascular diseases (asthma, high blood pressure, coronary heart disease, heart attack, and stroke). Using PM (sub 2.5) data and health data from the Centers for

  16. A Simple Visual Ethanol Biosensor Based on Alcohol Oxidase Immobilized onto Polyaniline Film for Halal Verification of Fermented Beverage Samples

    PubMed Central

    Kuswandi, Bambang; Irmawati, Titi; Hidayat, Moch Amrun; Jayus; Ahmad, Musa

    2014-01-01

    A simple visual ethanol biosensor based on alcohol oxidase (AOX) immobilised onto polyaniline (PANI) film for halal verification of fermented beverage samples is described. This biosensor responds to ethanol via a colour change from green to blue, due to the enzymatic reaction of ethanol that produces acetaldehyde and hydrogen peroxide, when the latter oxidizes the PANI film. The procedure to obtain this biosensor consists of the immobilization of AOX onto PANI film by adsorption. For the immobilisation, an AOX solution is deposited on the PANI film and left at room temperature until dried (30 min). The biosensor was constructed as a dip stick for visual and simple use. The colour changes of the films have been scanned and analysed using image analysis software (i.e., ImageJ) to study the characteristics of the biosensor's response toward ethanol. The biosensor has a linear response in an ethanol concentration range of 0.01%–0.8%, with a correlation coefficient (r) of 0.996. The limit detection of the biosensor was 0.001%, with reproducibility (RSD) of 1.6% and a life time up to seven weeks when stored at 4 °C. The biosensor provides accurate results for ethanol determination in fermented drinks and was in good agreement with the standard method (gas chromatography) results. Thus, the biosensor could be used as a simple visual method for ethanol determination in fermented beverage samples that can be useful for Muslim community for halal verification. PMID:24473284

  17. A simple visual ethanol biosensor based on alcohol oxidase immobilized onto polyaniline film for halal verification of fermented beverage samples.

    PubMed

    Kuswandi, Bambang; Irmawati, Titi; Hidayat, Moch Amrun; Jayus; Ahmad, Musa

    2014-01-27

    A simple visual ethanol biosensor based on alcohol oxidase (AOX) immobilised onto polyaniline (PANI) film for halal verification of fermented beverage samples is described. This biosensor responds to ethanol via a colour change from green to blue, due to the enzymatic reaction of ethanol that produces acetaldehyde and hydrogen peroxide, when the latter oxidizes the PANI film. The procedure to obtain this biosensor consists of the immobilization of AOX onto PANI film by adsorption. For the immobilisation, an AOX solution is deposited on the PANI film and left at room temperature until dried (30 min). The biosensor was constructed as a dip stick for visual and simple use. The colour changes of the films have been scanned and analysed using image analysis software (i.e., ImageJ) to study the characteristics of the biosensor's response toward ethanol. The biosensor has a linear response in an ethanol concentration range of 0.01%-0.8%, with a correlation coefficient (r) of 0.996. The limit detection of the biosensor was 0.001%, with reproducibility (RSD) of 1.6% and a life time up to seven weeks when stored at 4 °C. The biosensor provides accurate results for ethanol determination in fermented drinks and was in good agreement with the standard method (gas chromatography) results. Thus, the biosensor could be used as a simple visual method for ethanol determination in fermented beverage samples that can be useful for Muslim community for halal verification.

  18. Bioconversion of Airborne Methylamine by Immobilized Recombinant Amine Oxidase from the Thermotolerant Yeast Hansenula polymorpha

    PubMed Central

    Sigawi, Sasi; Nitzan, Yeshayahu

    2014-01-01

    Aliphatic amines, including methylamine, are air-pollutants, due to their intensive use in industry and the natural degradation of proteins, amino acids, and other nitrogen-containing compounds in biological samples. It is necessary to develop systems for removal of methylamine from the air, since airborne methylamine has a negative effect on human health. The primary amine oxidase (primary amine : oxygen oxidoreductase (deaminating) or amine oxidase, AMO; EC 1.4.3.21), a copper-containing enzyme from the thermotolerant yeast Hansenula polymorpha which was overexpressed in baker's yeast Saccharomyces cerevisiae, was tested for its ability to oxidize airborne methylamine. A continuous fluidized bed bioreactor (CFBR) was designed to enable bioconversion of airborne methylamine by AMO immobilized in calcium alginate (CA) beads. The results demonstrated that the bioreactor with immobilized AMO eliminates nearly 97% of the airborne methylamine. However, the enzymatic activity of AMO causes formation of formaldehyde. A two-step bioconversion process was therefore proposed. In the first step, airborne methylamine was fed into a CFBR which contained immobilized AMO. In the second step, the gas flow was passed through another CFBR, with alcohol oxidase from the yeast H. polymorpha immobilized in CA, in order to decompose the formaldehyde formed in the first step. The proposed system provided almost total elimination of the airborne methylamine and the formaldehyde. PMID:24672387

  19. Group motivational interviewing for adolescents: Change talk and alcohol and marijuana outcomes

    PubMed Central

    D’Amico, Elizabeth J.; Houck, Jon M.; Hunter, Sarah B.; Miles, Jeremy N.V.; Osilla, Karen Chan; Ewing, Brett A.

    2014-01-01

    Objective Little is known about what may distinguish effective and ineffective group interventions. Group motivational interviewing (MI) is a promising intervention for adolescent alcohol and other drug (AOD) use; however, the mechanisms of change for group MI are unknown. One potential mechanism is change talk, which is client speech arguing for change. The present study describes the group process in adolescent group MI and effects of group-level change talk on individual alcohol and marijuana outcomes. Method We analyzed 129 group session audio recordings from a randomized clinical trial of adolescent group MI. Sequential coding was performed using the Motivational Interviewing Skill Code (MISC) and the CASAA Application for Coding Treatment Interactions (CACTI) software application. Outcomes included past-month intentions, frequency, and consequences of alcohol and marijuana use, motivation to change, and positive expectancies. Results Sequential analysis indicated that facilitator open-ended questions and reflections of change talk (CT) increased group CT. Group CT was then followed by more CT. Multilevel models accounting for rolling group enrollment revealed group CT was associated with decreased alcohol intentions, alcohol use and heavy drinking three months later; group sustain talk was associated with decreased motivation to change, increased intentions to use marijuana, and increased positive alcohol and marijuana expectancies. Conclusions Facilitator speech and peer responses each had effects on change and sustain talk in the group setting, which was then associated with individual changes. Selective reflection of CT in adolescent group MI is suggested as a strategy to manage group dynamics and increase behavioral change. PMID:25365779

  20. Cultural Identity among Urban American Indian/Native Alaskan Youth: Implications for Alcohol and Drug Use

    PubMed Central

    Brown, Ryan A.; Dickerson, Daniel L.; D’Amico, Elizabeth J.

    2016-01-01

    American Indian and Alaska Native (AI/AN) youth exhibit high rates of alcohol and other drug (AOD) use, which is often linked to the social and cultural upheaval experienced by AI/ANs during the colonization of North America. Urban AI/AN youth may face unique challenges, including increased acculturative stress due to lower concentrations of AI/AN populations in urban areas. Few existing studies have explored cultural identity among urban AI/AN youth and its association with AOD use. Objectives This study used systematic qualitative methods with AI/AN communities in two urban areas within California to shed light on how urban AI/AN youth construct cultural identity and how this relates to AOD use and risk behaviors. Methods We conducted 10 focus groups with a total of 70 youth, parents, providers, and Community Advisory Board members and used team-based structured thematic analysis in the Dedoose software platform. Results We identified 12 themes: intergenerational stressors, cultural disconnection, AI/AN identity as protective, pan-tribal identity, mixed racial-ethnic identity, rural vs. urban environments, the importance of AI/AN institutions, stereotypes and harassment, cultural pride, developmental trajectories, risks of being AI/AN, and mainstream culture clash. Overall, youth voiced curiosity about their AI/AN roots and expressed interest in deepening their involvement in cultural activities. Adults described the myriad ways in which involvement in cultural activities provides therapeutic benefits for AI/AN youth. Conclusions Interventions that provide urban AI/AN youth with an opportunity to engage in cultural activities and connect with positive and healthy constructs in AI/AN culture may provide added impact to existing interventions. PMID:27450682

  1. H2O2 recycling during oxidation of the arylglycerol beta-aryl ether lignin structure by lignin peroxidase and glyoxal oxidase.

    PubMed

    Hammel, K E; Mozuch, M D; Jensen, K A; Kersten, P J

    1994-11-15

    Oxidative C alpha-C beta cleavage of the arylglycerol beta-aryl ether lignin model 1-(3,4-dimethoxy-phenyl)-2-phenoxypropane-1,3-diol (I) by Phanerochaete chrysosporium lignin peroxidase in the presence of limiting H2O2 was enhanced 4-5-fold by glyoxal oxidase from the same fungus. Further investigation showed that each C alpha-C beta cleavage reaction released 0.8-0.9 equiv of glycolaldehyde, a glyoxal oxidase substrate. The identification of glycolaldehyde was based on 13C NMR spectrometry of reaction product obtained from beta-, gamma-, and beta,gamma-13C-substituted I, and quantitation was based on an enzymatic NADH-linked assay. The oxidation of glycolaldehyde by glyoxal oxidase yielded 0.9 oxalate and 2.8 H2O2 per reaction, as shown by quantitation of oxalate as 2,3-dihydroxyquinoxaline after derivatization with 1,2-diaminobenzene and by quantitation of H2O2 in coupled spectrophotometric assays with veratryl alcohol and lignin peroxidase. These results suggest that the C alpha-C beta cleavage of I by lignin peroxidase in the presence of glyoxal oxidase should regenerate as many as 3 H2O2. Calculations based on the observed enhancement of LiP-catalyzed C alpha-C beta cleavage by glyoxal oxidase showed that approximately 2 H2O2 were actually regenerated per cleavage of I when both enzymes were present. The cleavage of arylglycerol beta-aryl ether structures by ligninolytic enzymes thus recycles H2O2 to support subsequent cleavage reactions.

  2. A new amperometric enzyme electrode for alcohol determination.

    PubMed

    Gülce, H; Gülce, A; Kavanoz, M; Coşkun, H; Yildiz, A

    2002-06-01

    A new enzyme electrode for the determination of alcohols was developed by immobilizing alcohol oxidase in polvinylferrocenium matrix coated on a Pt electrode surface. The amperometric response due to the electrooxidation of enzymatically generated H(2)O(2) was measured at a constant potential of +0.70 V versus SCE. The effects of substrate, buffer and enzyme concentrations, pH and temperature on the response of the electrode were investigated. The optimum pH was found to be pH 8.0 at 30 degrees C. The steady-state current of this enzyme electrode was reproducible within +/-5.0% of the relative error. The sensitivity of the enzyme electrode decreased in the following order: methanol>ethanol>n-butanol>benzyl alcohol. The linear response was observed up to 3.7 mM for methanol, 3.0 mM for ethanol, 6.2 mM for n-butanol, and 5.2 mM for benzyl alcohol. The apparent Michaelis-Menten constant (K(Mapp)) value and the activation energy, E(a), of this immobilized enzyme system were found to be 5.78 mM and 38.07 kJ/mol for methanol, respectively.

  3. Quantitation of immunoadsorbed flavoprotein oxidases by luminol-mediated chemiluminescence.

    PubMed

    Hinkkanen, A; Maly, F E; Decker, K

    1983-04-01

    The detection of the flavoenzymes 6-hydroxy-L-nicotine oxidase and 6-hydroxy-D-nicotine oxidase at the sub-femtomol level was achieved by coupling the reaction of the immunoadsorbed proteins to the peroxidase-catalysed oxidation of luminol. The H2O2-producing oxidases retained their full activity when bound to the respective immobilized antibodies. This fact allowed the concentration of the enzymes from very dilute solutions and the quantitative assay of their activities in the microU range. Due to strict stereoselectivity and the absence of immunological cross-reactivity, the two flavoproteins could be determined in the same solution. This method was used to measure the 6-hydroxy-D-nicotine oxidase and 6-hydroxy-L-nicotine oxidase activities in Escherichia coli RR1 and different Arthrobacter strains cultured under non-inducing conditions. The same activity ratio of 6-hydroxy-L-nicotine oxidase/6-hydroxy-D-nicotine oxidase as in D L-nicotine-induced cells of A. oxidans was observed in non-induced wild type and in riboflavin-requiring (rf-) mutant cells of this aerob.

  4. Regional characteristics of the relationship between columnar AOD and surface PM2.5: Application of lidar aerosol extinction profiles over Baltimore-Washington Corridor during DISCOVER-AQ

    NASA Astrophysics Data System (ADS)

    Chu, D. Allen; Ferrare, Richard; Szykman, James; Lewis, Jasper; Scarino, Amy; Hains, Jennifer; Burton, Sharon; Chen, Gao; Tsai, Tzuchin; Hostetler, Chris; Hair, Johnathan; Holben, Brent; Crawford, James

    2015-01-01

    The first field campaign of DISCOVER-AQ (Deriving Information on Surface conditions from COlumn and VERtically resolved observations relevant to Air Quality) took place in July 2011 over Baltimore-Washington Corridor (BWC). A suite of airborne remote sensing and in-situ sensors was deployed along with ground networks for mapping vertical and horizontal distribution of aerosols. Previous researches were based on a single lidar station because of the lack of regional coverage. This study uses the unique airborne HSRL (High Spectral Resolution Lidar) data to baseline PM2.5 (particulate matter of aerodynamic diameter less than 2.5 μm) estimates and applies to regional air quality with satellite AOD (Aerosol Optical Depth) retrievals over BWC (∼6500 km2). The linear approximation takes into account aerosols aloft above AML (Aerosol Mixing Layer) by normalizing AOD with haze layer height (i.e., AOD/HLH). The estimated PM2.5 mass concentrations by HSRL AOD/HLH are shown within 2 RMSE (Root Mean Square Error ∼9.6 μg/m3) with correlation ∼0.88 with the observed over BWC. Similar statistics are shown when applying HLH data from a single location over the distance of 100 km. In other words, a single lidar is feasible to cover the range of 100 km with expected uncertainties. The employment of MPLNET-AERONET (MicroPulse Lidar NETwork - AErosol RObotic NETwork) measurements at NASA GSFC produces similar statistics of PM2.5 estimates as those derived by HSRL. The synergy of active and passive remote sensing aerosol measurements provides the foundation for satellite application of air quality on a daily basis. For the optimal range of 10 km, the MODIS-estimated PM2.5 values are found satisfactory at 27 (out of 36) sunphotometer locations with mean RMSE of 1.6-3.3 μg/m3 relative to PM2.5 estimated by sunphotometers. The remaining 6 of 8 marginal sites are found in the coastal zone, for which associated large RMSE values ∼4.5-7.8 μg/m3 are most likely due to

  5. Role of NADPH oxidases and reactive oxygen species in regulation of bone turnover and the skeletal toxicity of alcohol

    USDA-ARS?s Scientific Manuscript database

    Recent studies with genetically modified mice and dietary antioxidants have suggested an important role for superoxide derived from NADPH oxidase (NOX) enzymes and other reactive oxygen species (ROS) such as hydrogen peroxide in regulation of normal bone turnover during development and also in the r...

  6. Influence of gamma irradiation on polymerization of pyrrole and glucose oxidase immobilization onto poly (pyrrole)/poly (vinyl alcohol) matrix

    NASA Astrophysics Data System (ADS)

    Idris, Sarada; A. Bakar, Ahmad Ashrif; Thevy Ratnam, Chantara; Kamaruddin, Nur Hasiba; Shaari, Sahbudin

    2017-04-01

    This paper describes the immobilization of glucose oxidase, GOx onto polymer matrix comprising of poly(pyrrole), PPy and poly(vinyl alcohol), PVA using gamma irradiation technique. Py/PVA-GOx film was prepared by spreading PVA:GOx, 1:1 solution onto dried pyrrole film and exposed to gamma irradiation from cobalt 60 source at doses ranging from 0 to 60 kGy. The films were subjected to structural and morphological analyses by using Fourier transform infrared spectroscopy (FTIR), X-ray photoelectron spectroscopy (XPS), Scanning electron microscope (SEM), Field emission scanning electron microscope (FESEM) and Atomic-force microscopy (AFM) techniques. Similar studies were also made on pristine pyrrole film which served as control. The SEM and FTIR spectra of Py/PVA-GOx film revealed that pyrrole has been successfully polymerized through irradiation-induced reactions. The results on the morphological properties of the samples characterize using FESEM, SEM and AFM further confirmed the occurrence of radiation-induced modification of Py/PVA-GOx film. The FTIR spectra showed the existence of intermolecular interaction between polymer matrix and GOx indicating that GOx had been successfully immobilized onto Ppy/PVA matrix by radiation-induced reactions. Results revealed that radiation induced reactions such as polymerization of pyrrole, crosslinking of PVA, grafting between the adjacent PVA and pyrrole molecules as well as immobilization of GOx onto Ppy/PVA matrix occurred simultaneously upon gamma irradiation. The optimum dose for GOx immobilization in the polymer matrix found to be 40 kGy. Therefore it is clear that this irradiation technique offered a simple single process to produce Py/PVA-GOx film without additional crosslinking and polymerization agents.

  7. Correlation Between Monoamine Oxidase Inhibitors and Anticonvulsants

    PubMed Central

    Dwivedi, Chandradhar; Misra, Radhey S.; Chaudhari, Anshumali; Parmar, Surendra S.

    1980-01-01

    Monoamine oxidase inhibitory and anticonvulsant properties of 2-substituted styryl-6-bromo-3-(4-ethylbenzoate/4 benzhydrazide)-4-quinazoles are studied. All styryl quinazolone esters except compound number 9 exhibited monoamine oxidase inhibitory properties during oxidative deamination of kynuramine. Corresponding hydrazides were found to have relatively higher activity. All these quinazolones were able to protect against pentylenetetrazol induced seizures. These observations in general do not prove that monoamine oxidase inhibitory properties represent the biochemical basis for the anticonvulsant activity of these compounds. PMID:7420438

  8. Regulation of tyramine oxidase synthesis in Klebsiella aerogenes.

    PubMed Central

    Okamura, H; Murooka, Y; Harada, T

    1976-01-01

    Tyramine oxidase in Klebsiella aerogenes is highly specific for tyramine, dopamine, octopamine, and norepinephrine, and its synthesis is induced specifically by these compounds. The enzyme is present in a membrane-bound form. The Km value for tyramine is 9 X 10(-4) M. Tyramine oxidase synthesis was subjected to catabolite repression by glucose in the presence of ammonium salts. Addition of cyclic adenosine 3',5'-monophosphate (cAMP) overcame the catabolite repression. A mutant strain, K711, which can produce a high level of beta-galactosidase in the presence of glucose and ammonium chloride, can also synthesize tyramine oxidase and histidase in the presence of inducer in glucose ammonium medium. Catabolite repression of tyramine oxidase synthesis was relieved when the cells were grown under conditions of nitrogen limitation, whereas beta-galactosidase was strongly repressed under these conditions. A cAMP-requiring mutant, MK54, synthesized tyramine oxidase rapidly when tyramine was used as the sole source of nitrogen in the absence of cAMP. However, a glutamine synthetase-constitutive mutant, MK94, failed to synthesize tyramine oxidase in the presence of glucose and ammonium chloride, although it synthesized histidase rapidly under these conditions. These results suggest that catabolite repression of tyramine oxidase synthesis in K. aerogenes is regulated by the intracellular level of cAMP and an unknown cytoplasmic factor that acts independently of cAMP and is formed under conditions of nitrogen limitation. PMID:179974

  9. Identifying Optimal Temporal Scale for the Correlation of AOD and Ground Measurements of PM2.5 to Improve the Model Performance in a Real-time Air Quality Estimation System

    NASA Technical Reports Server (NTRS)

    Li, Hui; Faruque, Fazlay; Williams, Worth; Al-Hamdan, Mohammad; Luvall, Jeffrey C.; Crosson, William; Rickman, Douglas; Limaye, Ashutosh

    2009-01-01

    Aerosol optical depth (AOD), an indirect estimate of particle matter using satellite observations, has shown great promise in improving estimates of PM 2.5 air quality surface. Currently, few studies have been conducted to explore the optimal way to apply AOD data to improve the model accuracy of PM 2.5 surface estimation in a real-time air quality system. We believe that two major aspects may be worthy of consideration in that area: 1) the approach to integrate satellite measurements with ground measurements in the pollution estimation, and 2) identification of an optimal temporal scale to calculate the correlation of AOD and ground measurements. This paper is focused on the second aspect on the identifying the optimal temporal scale to correlate AOD with PM2.5. Five following different temporal scales were chosen to evaluate their impact on the model performance: 1) within the last 3 days, 2) within the last 10 days, 3) within the last 30 days, 4) within the last 90 days, and 5) the time period with the highest correlation in a year. The model performance is evaluated for its accuracy, bias, and errors based on the following selected statistics: the Mean Bias, the Normalized Mean Bias, the Root Mean Square Error, Normalized Mean Error, and the Index of Agreement. This research shows that the model with the temporal scale of within the last 30 days displays the best model performance in this study area using 2004 and 2005 data sets.

  10. Monoamine Oxidase A Promoter Variable Number of Tandem Repeats (MAOA-uVNTR) in Alcoholics According to Lesch Typology

    PubMed Central

    Samochowiec, Agnieszka; Chęć, Magdalena; Kopaczewska, Edyta; Samochowiec, Jerzy; Lesch, Otto; Grochans, Elżbieta; Jasiewicz, Andrzej; Bienkowski, Przemyslaw; Łukasz, Kołodziej; Grzywacz, Anna

    2015-01-01

    Background: The aim of this study was to examine the association between the MAOA-uVNTR gene polymorphism in a homogeneous subgroups of patients with alcohol dependence categorized according to Lesch’s typology. Methods: DNA was provided from alcohol dependent (AD) patients (n = 370) and healthy control subjects (n = 168) all of Polish descent. The history of alcoholism was obtained using the Polish version of the Semi-Structured Assessment for the Genetics of Alcoholism (SSAGA). Samples were genotyped using PCR methods. Results: We found no association between alcohol dependence and MAOA gene polymorphism. Conclusions: Lesch typology is a clinical consequence of the disease and its phenotypic description is too complex for a simple genetic analysis. PMID:25809512

  11. Cell phone-based ecological momentary assessment of substance use context for Latino youth in outpatient treatment: Who, what, when and where.

    PubMed

    Comulada, W Scott; Swendeman, Dallas; Wu, Nancy

    2016-10-01

    Relationships between alcohol, marijuana and other drug (AOD) use and contextual factors have mostly been established through retrospective self-report. Given the embeddedness of cell phones in adolescents' daily activities, cell phone-based ecological momentary assessment (CEMA) provides an opportunity to better understand AOD use in youth and how cell phones can be used to self-monitor and deliver interventions. We use CEMA to examine AOD use in Latino youth who have been especially understudied. Twenty-eight mostly Latino youth (ages 13-18) in outpatient substance abuse treatment recorded AOD use, contextual factors, cravings, and affect through once-daily CEMA over one month periods. Random-effects logistic regression was used to compare contextual factors between periods of AOD use and non-use. The most frequent contextual factors reported during AOD use were being with close friends and "hanging out" as the primary activity. During AOD use compared to non-use, youth were more likely to be with close friends (OR=4.76; p<0.01), around users (OR=17.69; p<0.01), and at a friend's house (OR=5.97; p<0.01). Alcohol use was more frequently reported at night (63% vs 34%) and on weekends relative to other substances (64% vs 49%). Strong cravings were more frequently reported on AOD-use days (OR=7.34; p<0.01). Types of positive and negative affect were reported with similar frequencies, regardless of AOD use. Reporting on social context, location, day and time of day, and cravings all show promise in developing cell phone-based interventions triggered by contextual data. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

  12. NADPH oxidases of the brain: distribution, regulation, and function.

    PubMed

    Infanger, David W; Sharma, Ram V; Davisson, Robin L

    2006-01-01

    The NADPH oxidase is a multi-subunit enzyme that catalyzes the reduction of molecular oxygen to form superoxide (O(2)(-)). While classically linked to the respiratory burst in neutrophils, recent evidence now shows that O(2)(-) (and associated reactive oxygen species, ROS) generated by NADPH oxidase in nonphagocytic cells serves myriad functions in health and disease. An entire new family of NADPH Oxidase (Nox) homologues has emerged, which vary widely in cell and tissue distribution, as well as in function and regulation. A major concept in redox signaling is that while NADPH oxidase-derived ROS are necessary for normal cellular function, excessive oxidative stress can contribute to pathological disease. This certainly is true in the central nervous system (CNS), where normal NADPH oxidase function appears to be required for processes such as neuronal signaling, memory, and central cardiovascular homeostasis, but overproduction of ROS contributes to neurotoxicity, neurodegeneration, and cardiovascular diseases. Despite implications of NADPH oxidase in normal and pathological CNS processes, still relatively little is known about the mechanisms involved. This paper summarizes the evidence for NADPH oxidase distribution, regulation, and function in the CNS, emphasizing the diversity of Nox isoforms and their new and emerging role in neuro-cardiovascular function. In addition, perspectives for future research and novel therapeutic targets are offered.

  13. Gravity Responsive NADH Oxidase of the Plasma Membrane

    NASA Technical Reports Server (NTRS)

    Morre, D. James (Inventor)

    2002-01-01

    A method and apparatus for sensing gravity using an NADH oxidase of the plasma membrane which has been found to respond to unit gravity and low centrifugal g forces. The oxidation rate of NADH supplied to the NADH oxidase is measured and translated to represent the relative gravitational force exerted on the protein. The NADH oxidase of the plasma membrane may be obtained from plant or animal sources or may be produced recombinantly.

  14. NADPH Oxidase Plays a Role on Ethanol-Induced Hypertension and Reactive Oxygen Species Generation in the Vasculature.

    PubMed

    Marchi, Katia Colombo; Ceron, Carla Speroni; Muniz, Jaqueline J; De Martinis, Bruno S; Tanus-Santos, José E; Tirapelli, Carlos Renato

    2016-09-01

    Investigate the role of NADPH oxidase on ethanol-induced hypertension and vascular oxidative stress. Male Wistar rats were treated with ethanol (20% v/v). Apocynin (10 mg/kg/day, i.p.) prevented ethanol-induced hypertension. The increased contractility of endothelium-intact and endothelium-denuded aortic rings from ethanol-treated rats to phenylephrine was prevented by apocynin. Ethanol consumption increased superoxide anion (O2 (-)) generation and lipid peroxidation and apocynin prevented these responses. The decrease on plasma and vascular nitrate/nitrite (NOx) levels induced by ethanol was not prevented by apocynin. Treatment with ethanol did not affect aortic levels of hydrogen peroxide (H2O2) or reduced glutathione (GSH). Ethanol did not alter the activities of xanthine oxidase (XO), superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx). Ethanol increased the expression of Nox1, PKCδ, nNOS, SAPK/JNK and SOD2 in the rat aorta and apocynin prevented these responses. No difference on aortic expression of Nox2, Nox4, p47phox, Nox organizer 1 (Noxo1), eNOS and iNOS was detected after treatment with ethanol. Ethanol treatment did not alter the phosphorylation of SAPK/JNK, p38MAPK, c-Src, Rac1 or PKCδ. The major new finding of our study is that the increased vascular generation of reactive oxygen species (ROS) induced by ethanol is related to increased vascular Nox1/NADPH oxidase expression. This mechanism is involved in vascular dysfunction and hypertension induced by ethanol. Additionally, we conclude that ethanol consumption induces the expression of different proteins that regulate vascular contraction and growth and that NADPH oxidase-derived ROS play a role in such response. The key findings of our study are that ethanol-induced hypertension is mediated by NADPH oxidase. Moreover, increased vascular Nox1 expression is related to the generation of reactive oxygen species (ROS) by ethanol. Finally, ROS induced by ethanol increase the

  15. Strategies to facilitate integrated care for people with alcohol and other drug problems: a systematic review.

    PubMed

    Savic, Michael; Best, David; Manning, Victoria; Lubman, Dan I

    2017-04-07

    There is a growing body of research highlighting the potential benefits of integrated care as a way of addressing the needs of people with alcohol and other drug (AOD) problems, given the broad range of other issues clients often experience. However, there has been little academic attention on the strategies that treatment systems, agencies and clinicians could implement to facilitate integrated care. We synthesised the existing evidence on strategies to improve integrated care in an AOD treatment context by conducting a systematic review of the literature. We searched major academic databases for peer-reviewed articles that evaluated strategies that contribute to integrated care in an AOD context between 1990 and 2014. Over 2600 articles were identified, of which 14 met the study inclusion criteria of reporting on an empirical study to evaluate the implementation of integrated care strategies. The types of strategies utilised in included articles were then synthesised. We identified a number of interconnected strategies at the funding, organisational, service delivery and clinical levels. Ensuring that integrated care is included within service specifications of commissioning bodies and is adequately funded was found to be critical in effective integration. Cultivating positive inter-agency relationships underpinned and enabled the implementation of most strategies identified. Staff training in identifying and responding to needs beyond clinicians' primary area of expertise was considered important at a service level. However, some studies highlight the need to move beyond discrete training events and towards longer term coaching-type activities focussed on implementation and capacity building. Sharing of client information (subject to informed consent) was critical for most integrated care strategies. Case-management was found to be a particularly good approach to responding to the needs of clients with multiple and complex needs. At the clinical level, screening

  16. Effects of MAOA-Genotype, Alcohol Consumption, and Aging on Violent Behavior

    PubMed Central

    Tikkanen, Roope; Sjöberg, Rickard L.; Ducci, Francesca; Goldman, David; Holi, Matti; Tiihonen, Jari; Virkkunen, Matti

    2009-01-01

    Background Environmental factors appear to interact with a functional polymorphism (MAOA-LPR) in the promoter region of the monoamine oxidase A gene (MAOA) in determining some forms of antisocial behavior. However, how MAOA-LPR modulates the effects of other factors such as alcohol consumption related to antisocial behavior is not completely understood. Methods This study examines the conjunct effect of MAOA-LPR, alcohol consumption, and aging on the risk for violent behavior. Recidivism in severe impulsive violent behavior was assessed after 7 to 15 years in a sample of 174 Finnish alcoholic offenders, the majority of whom exhibited antisocial or borderline personality disorder or both, and featured impulsive temperament traits. Results The risk for committing new acts of violence increased by 2.3% for each kilogram of increase in yearly mean alcohol consumption (p = 0.004) and decreased by 7.3% for every year among offenders carrying the high activity MAOA genotype. In contrast, alcohol consumption and aging failed to affect violent behavior in the low activity MAOA genotyped offenders. MAOA-LPR showed no main effect on the risk for recidivistic violence. Conclusions Violent offenders carrying the high activity MAOA genotype differ in several ways from carriers with the low activity MAOA risk allele previously associated with antisocial behavior. Finnish high activity MAOA genotyped risk alcoholics exhibiting antisocial behavior, high alcohol consumption, and abnormal alcohol-related impulsive and uncontrolled violence might represent an etiologically distinct alcohol dependence subtype. PMID:19120058

  17. Effects of MAOA-genotype, alcohol consumption, and aging on violent behavior.

    PubMed

    Tikkanen, Roope; Sjöberg, Rickard L; Ducci, Francesca; Goldman, David; Holi, Matti; Tiihonen, Jari; Virkkunen, Matti

    2009-03-01

    Environmental factors appear to interact with a functional polymorphism (MAOA-LPR) in the promoter region of the monoamine oxidase A gene (MAOA) in determining some forms of antisocial behavior. However, how MAOA-LPR modulates the effects of other factors such as alcohol consumption related to antisocial behavior is not completely understood. This study examines the conjunct effect of MAOA-LPR, alcohol consumption, and aging on the risk for violent behavior. Recidivism in severe impulsive violent behavior was assessed after 7 to 15 years in a sample of 174 Finnish alcoholic offenders, the majority of whom exhibited antisocial or borderline personality disorder or both, and featured impulsive temperament traits. The risk for committing new acts of violence increased by 2.3% for each kilogram of increase in yearly mean alcohol consumption (p = 0.004) and decreased by 7.3% for every year among offenders carrying the high activity MAOA genotype. In contrast, alcohol consumption and aging failed to affect violent behavior in the low activity MAOA genotyped offenders. MAOA-LPR showed no main effect on the risk for recidivistic violence. Violent offenders carrying the high activity MAOA genotype differ in several ways from carriers with the low activity MAOA risk allele previously associated with antisocial behavior. Finnish high activity MAOA genotyped risk alcoholics exhibiting antisocial behavior, high alcohol consumption, and abnormal alcohol-related impulsive and uncontrolled violence might represent an etiologically distinct alcohol dependence subtype.

  18. Serotonin transporter promoter polymorphism and monoamine oxidase type A VNTR allelic variants together influence alcohol binge drinking risk in young women.

    PubMed

    Herman, Aryeh I; Kaiss, Kristi M; Ma, Rui; Philbeck, John W; Hasan, Asfar; Dasti, Humza; DePetrillo, Paolo B

    2005-02-05

    The short allelic variant of the serotonin transporter protein promoter polymorphism (5HTTLPR) appears to influence binge drinking in college students. Both monoamine oxidase type A (MAOA) and the serotonin transporter protein are involved in the processing of serotonin, and allelic variants are both associated with differences in the efficiency of expression. We hypothesized that a significant gene x gene interaction would further stratify the risk of binge drinking in this population. Participants were college students (n = 412) who completed the College Alcohol Study, used to measure binge drinking behaviors. Genomic DNA was extracted from saliva for PCR based genotyping. The risk function for binge drinking was modeled using logistic regression, with final model fit P < 0.0005. This model was valid only for Caucasian females (n = 223), but the power to detect sex and ethnic effects was small. Young Caucasian women carrying higher expression MAOA VNTR alleles homozygous for the short allelic variant of the 5HTTLPR demonstrated the highest rate of binge drinking by self-report, odds ratio (genotype odds: population odds) and 95% confidence intervals, 3.11 (1.14-18.10). Individuals carrying higher expression MAOA VNTR alleles carrying at least one long 5HTTLPR allelic variant had the lowest risk of binge drinking 0.46 (0.28-0.71). These results support the hypothesis that binge drinking behavior in young adulthood may be influenced by neurobiological differences in serotonergic function conferred by functional polymorphisms in genes involved in serotonin processing. (c) 2004 Wiley-Liss, Inc.

  19. Calcium transport in vesicles energized by cytochrome oxidase

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

    Rosier, Randy N.

    1979-01-01

    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 interactionmore » 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.« less

  20. Possible interaction between MAOA and DRD2 genes associated with antisocial alcoholism among Han Chinese men in Taiwan.

    PubMed

    Wang, Tso-Jen; Huang, San-Yuan; Lin, Wei-Wen; Lo, Hsin-Yi; Wu, Pei-Lin; Wang, Yu-Shan; Wu, Yi-Syuan; Ko, Huei-Chen; Shih, Jean-Chen; Lu, Ru-Band

    2007-01-30

    Both monoamine oxidase A (MAOA) and dopamine D(2) receptor (DRD2) genes have been considered as candidate genes for antisocial personality disorder with alcoholism (Antisocial ALC) [Parsian, A., 1999. Sequence analysis of exon 8 of MAO-A gene in alcoholics with antisocial personality and normal controls. Genomics. 45, 290-295.; Samochowiec, J., Lesch, K.P., Rottmann, M., Smolka, M., Syagailo, Y.V., Okladnova, O., Rommelspacher, H., Winterer, G., Schmidt, L.G., Sander, T., 1999. Association of a regulatory polymorphism in the promoter region of the monoamine oxidase A gene with antisocial alcoholism. Psychiatry. Res. 86, 67-72.; Schmidt, L.vG., Sander, T., Kuhn, S., Smolka, M., Rommelspacher, H., Samochowiec, J., Lesch, K.P., 2000. Different allele distribution of a regulatory MAO-A gene promotor polymorphism in antisocial and anxious-depressive alcoholics. J. Neural .Transm. 107, 681-689.]. However, the association between alcoholism and MAOA or DRD2 gene has not been universally accepted [Lee, J.F., Lu, R.B., Ko, H.C., Chang, F.M., Yin, S.J., Pakstis, A.J., Kidd, K.K., 1999. No association between DRD(2) locus and alcoholism after controlling the ADH and ALDH genotypes in Chinese Han population. Alcohol. Clin. Exp. Res. 23, 592-599.; Lu, R.B., Lin, W.W., Lee, J.F., Ko, H.C., Shih, J.C., 2003. Neither antisocial personality disorder nor antisocial alcoholism association with MAOA gene among Han Chinese males in Taiwan. Alcohol. Clin. Exp. Res. 27, 889-893.]. Since dopamine is metabolized to 3,4-dihydroxyphenyl-acetaldehyde (DOPAL) via monoamine oxidase (MAO) [Westerink, B.H., de Vries, J.B., 1985. On the origin of dopamine and its metabolite in predominantly noradrenergic innervated brain areas. Brain. Res. 330, 164-166.], the interaction between MAOA and DRD2 genes might be related to Antisocial ALC. The present study aimed to determine whether Antisocial ALC might be associated with the possible interactions of DRD2 gene with MAOA gene. Of the 231 Han Chinese

  1. Biomimetic enzyme nanocomplexes and their use as antidotes and preventive measures for alcohol intoxication

    NASA Astrophysics Data System (ADS)

    Liu, Yang; Du, Juanjuan; Yan, Ming; Lau, Mo Yin; Hu, Jay; Han, Hui; Yang, Otto O.; Liang, Sheng; Wei, Wei; Wang, Hui; Li, Jianmin; Zhu, Xinyuan; Shi, Linqi; Chen, Wei; Ji, Cheng; Lu, Yunfeng

    2013-03-01

    Organisms have sophisticated subcellular compartments containing enzymes that function in tandem. These confined compartments ensure effective chemical transformation and transport of molecules, and the elimination of toxic metabolic wastes. Creating functional enzyme complexes that are confined in a similar way remains challenging. Here we show that two or more enzymes with complementary functions can be assembled and encapsulated within a thin polymer shell to form enzyme nanocomplexes. These nanocomplexes exhibit improved catalytic efficiency and enhanced stability when compared with free enzymes. Furthermore, the co-localized enzymes display complementary functions, whereby toxic intermediates generated by one enzyme can be promptly eliminated by another enzyme. We show that nanocomplexes containing alcohol oxidase and catalase could reduce blood alcohol levels in intoxicated mice, offering an alternative antidote and prophylactic for alcohol intoxication.

  2. Substance Abuse among Juvenile Delinquents and Gang Members. Prevention Research Update Number Six, Spring 1990.

    ERIC Educational Resources Information Center

    Pollard, John A.; Austin, Gregory A.

    There is a strong statistical correlation between delinquency activity level and the level of alcohol and other drug (AOD) use in adolescents. A strong association between drug use, drug trafficking, and youth gangs has also emerged. However, several important questions concerning the relationship of delinquency, gang membership, and AOD use…

  3. Pacific oyster polyamine oxidase: a protein missing link in invertebrate evolution.

    PubMed

    Cervelli, Manuela; Polticelli, Fabio; Angelucci, Emanuela; Di Muzio, Elena; Stano, Pasquale; Mariottini, Paolo

    2015-05-01

    Polyamine oxidases catalyse the oxidation of polyamines and acetylpolyamines and are responsible for the polyamine interconversion metabolism in animal cells. Polyamine oxidases from yeast can oxidize spermine, N(1)-acetylspermine, and N(1)-acetylspermidine, while in vertebrates two different enzymes, namely spermine oxidase and acetylpolyamine oxidase, specifically catalyse the oxidation of spermine, and N(1)-acetylspermine/N(1)-acetylspermidine, respectively. In this work we proved that the specialized vertebrate spermine and acetylpolyamine oxidases have arisen from an ancestor invertebrate polyamine oxidase with lower specificity for polyamine substrates, as demonstrated by the enzymatic activity of the mollusc polyamine oxidase characterized here. This is the first report of an invertebrate polyamine oxidase, the Pacific oyster Crassostrea gigas (CgiPAO), overexpressed as a recombinant protein. This enzyme was biochemically characterized and demonstrated to be able to oxidase both N(1)-acetylspermine and spermine, albeit with different efficiency. Circular dichroism analysis gave an estimation of the secondary structure content and modelling of the three-dimensional structure of this protein and docking studies highlighted active site features. The availability of this pluripotent enzyme can have applications in crystallographic studies and pharmaceutical biotechnologies, including anticancer therapy as a source of hydrogen peroxide able to induce cancer cell death.

  4. Evolution of cytochrome oxidase, an enzyme older than atmospheric oxygen.

    PubMed

    Castresana, J; Lübben, M; Saraste, M; Higgins, D G

    1994-06-01

    Cytochrome oxidase is a key enzyme in aerobic metabolism. All the recorded eubacterial (domain Bacteria) and archaebacterial (Archaea) sequences of subunits 1 and 2 of this protein complex have been used for a comprehensive evolutionary analysis. The phylogenetic trees reveal several processes of gene duplication. Some of these are ancient, having occurred in the common ancestor of Bacteria and Archaea, whereas others have occurred in specific lines of Bacteria. We show that eubacterial quinol oxidase was derived from cytochrome c oxidase in Gram-positive bacteria and that archaebacterial quinol oxidase has an independent origin. A considerable amount of evidence suggests that Proteobacteria (Purple bacteria) acquired quinol oxidase through a lateral gene transfer from Gram-positive bacteria. The prevalent hypothesis that aerobic metabolism arose several times in evolution after oxygenic photosynthesis, is not sustained by two aspects of the molecular data. First, cytochrome oxidase was present in the common ancestor of Archaea and Bacteria whereas oxygenic photosynthesis appeared in Bacteria. Second, an extant cytochrome oxidase in nitrogen-fixing bacteria shows that aerobic metabolism is possible in an environment with a very low level of oxygen, such as the root nodules of leguminous plants. Therefore, we propose that aerobic metabolism in organisms with cytochrome oxidase has a monophyletic and ancient origin, prior to the appearance of eubacterial oxygenic photosynthetic organisms.

  5. Inhibition of Human Vascular NADPH Oxidase by Apocynin Derived Oligophenols

    PubMed Central

    Mora-Pale, Mauricio; Weïwer, Michel; Yu, Jingjing; Linhardt, Robert J.; Dordick, Jonathan S.

    2009-01-01

    Enzymatic oxidation of apocynin, which may mimic in vivo metabolism, affords a large number of oligomers (apocynin oxidation products, AOP) that inhibit vascular NADPH oxidase. In vitro studies of NADPH oxidase activity were performed to identify active inhibitors, resulting in a trimer hydroxylated quinone (IIIHyQ) that inhibited NADPH oxidase with an IC50 = 31 nM. Apocynin itself possessed minimal inhibitory activity. NADPH oxidase is believed to be inhibited through prevention of the interaction between two NADPH oxidase subunits, p47phox and p22phox. To that end, while apocynin was unable to block the interaction of his-tagged p47phox with a surface immobilized biotinalyted p22phox peptide, the IIIHyQ product strongly interfered with this interaction (apparent IC50 = 1.6 μM). These results provide evidence that peroxidase-catalyzed AOP, which consist of oligomeric phenols and quinones, inhibit critical interactions that are involved in the assembly and activation of human vascular NADPH oxidase. PMID:19523836

  6. Current status of NADPH oxidase research in cardiovascular pharmacology.

    PubMed

    Rodiño-Janeiro, Bruno K; Paradela-Dobarro, Beatriz; Castiñeiras-Landeira, María Isabel; Raposeiras-Roubín, Sergio; González-Juanatey, José R; Alvarez, Ezequiel

    2013-01-01

    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 and oral bioavailability. However, other possibilities are not closed, with peptide inhibitors or monoclonal antibodies against NADPH oxidase isoforms continuing to be under investigation as well as the ongoing search for naturally occurring compounds. Likewise, some different approaches include inhibition of assembly of the NADPH oxidase complex, subcellular translocation, post-transductional modifications, calcium entry/release, electron transfer, and genetic expression. High-throughput screens for any of these activities could provide new

  7. Current status of NADPH oxidase research in cardiovascular pharmacology

    PubMed Central

    Rodiño-Janeiro, Bruno K; Paradela-Dobarro, Beatriz; Castiñeiras-Landeira, María Isabel; Raposeiras-Roubín, Sergio; González-Juanatey, José R; Álvarez, Ezequiel

    2013-01-01

    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 and oral bioavailability. However, other possibilities are not closed, with peptide inhibitors or monoclonal antibodies against NADPH oxidase isoforms continuing to be under investigation as well as the ongoing search for naturally occurring compounds. Likewise, some different approaches include inhibition of assembly of the NADPH oxidase complex, subcellular translocation, post-transductional modifications, calcium entry/release, electron transfer, and genetic expression. High-throughput screens for any of these activities could provide new

  8. Amine oxidase-based biosensors for spermine and spermidine determination.

    PubMed

    Boffi, Alberto; Favero, Gabriele; Federico, Rodolfo; Macone, Alberto; Antiochia, Riccarda; Tortolini, Cristina; Sanzó, Gabriella; Mazzei, Franco

    2015-02-01

    The present work describes the development and optimization of electrochemical biosensors for specific determination of the biogenic polyamine spermine (Spm) and spermidine (Spmd) whose assessment represents a novel important analytical tool in food analysis and human diagnostics. These biosensors have been prepared using novel engineered enzymes: polyamine oxidase (PAO) endowed with selectivity towards Spm and Spmd and spermine oxidase (SMO) characterized by strict specificity towards Spm. The current design entails biosensors in which the enzymes were entrapped in poly(vinyl alcohol) bearing styrylpyridinium groups (PVA-SbQ), a photocrosslinkable gel, onto an electrode surface. Screen-printed electrodes (SPEs) were used as electrochemical transducers for enzymatically produced hydrogen peroxide, operating at different potential vs Ag/AgCl according to the material of the working electrode (WE): +700 mV for graphite (GP) or -100 mV for Prussian blue (PB)-modified SPE, respectively. Biosensor performances were evaluated by means of flow injection amperometric (FIA) measurements. The modified electrodes showed good sensitivity, long-term stability and reproducibility. Under optimal conditions, the PAO biosensor showed a linear range 0.003-0.3 mM for Spm and 0.01-0.4 mM for Spmd, while with the SMO biosensor, a linear range of 0.004-0.5 mM for Spm has been obtained. The main kinetic parameters apparent Michaelis constant (K M), turnover number (K cat) and steady-state current (I max) were determined. The proposed device was then applied to the determination of biogenic amines in blood samples. The results obtained were in good agreement with those obtained with the GC-MS reference method.

  9. Oxygen activation in flavoprotein oxidases: the importance of being positive.

    PubMed

    Gadda, Giovanni

    2012-04-03

    The oxidation of flavin hydroquinones by O(2) in solution is slow, with second-order rate constants of ~250 M(-1) s(-1). This is due to the obligatory, single-electron transfer that initiates the reaction being thermodynamically unfavored and poorly catalyzed. Notwithstanding considerations of O(2) accessibility to the reaction site, its desolvation and geometry and other factors that can also contribute to further rate acceleration, flavoprotein oxidases must activate O(2) for reaction with flavin hydroquinones to be able to achieve the 100-1000-fold rate enhancements typically observed. Protein positive charges have been identified in glucose oxidase, monomeric sarcosine oxidase, N-methyltryptophan oxidase and fructosamine oxidase that electrostatically stabilize the transition state for the initial single electron transfer that generates the O(2)(-•)/flavin semiquinone radical pair. In choline oxidase despite the presence of three histidines in the active site, the trimethylammonium group of the reaction product provides such an electrostatic stabilization. A nonpolar site proximal to the flavin C(4a) atom in choline oxidase has also been identified, which contributes to the geometry and desolvation of the O(2) reaction site. The relevance of O(2) activation by product charges to other flavoprotein oxidases, such as for example those catalyzing amine oxidations, is discussed in this review. A nonpolar site close to the flavin C(4a) atom and a positive charge is identified through structural analysis in several flavoprotein oxidases. Mutagenesis has disclosed nonpolar sites in O(2)-reducing enzymes that utilize copper/TPQ or iron. It is predicted that classes of O(2)-reducing enzymes utilizing other cofactors also contain a similar catalytic motif.

  10. The First Mammalian Aldehyde Oxidase Crystal Structure

    PubMed Central

    Coelho, Catarina; Mahro, Martin; Trincão, José; Carvalho, Alexandra T. P.; Ramos, Maria João; Terao, Mineko; Garattini, Enrico; Leimkühler, Silke; Romão, Maria João

    2012-01-01

    Aldehyde oxidases (AOXs) are homodimeric proteins belonging to the xanthine oxidase family of molybdenum-containing enzymes. Each 150-kDa monomer contains a FAD redox cofactor, two spectroscopically distinct [2Fe-2S] clusters, and a molybdenum cofactor located within the protein active site. AOXs are characterized by broad range substrate specificity, oxidizing different aldehydes and aromatic N-heterocycles. Despite increasing recognition of its role in the metabolism of drugs and xenobiotics, the physiological function of the protein is still largely unknown. We have crystallized and solved the crystal structure of mouse liver aldehyde oxidase 3 to 2.9 Å. This is the first mammalian AOX whose structure has been solved. The structure provides important insights into the protein active center and further evidence on the catalytic differences characterizing AOX and xanthine oxidoreductase. The mouse liver aldehyde oxidase 3 three-dimensional structure combined with kinetic, mutagenesis data, molecular docking, and molecular dynamics studies make a decisive contribution to understand the molecular basis of its rather broad substrate specificity. PMID:23019336

  11. CotA, a Multicopper Oxidase from Bacillus pumilus WH4, Exhibits Manganese-Oxidase Activity

    PubMed Central

    Su, Jianmei; Bao, Peng; Bai, Tenglong; Deng, Lin; Wu, Hui; Liu, Fan; He, Jin

    2013-01-01

    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 a

  12. Generating disulfides with the quiescin sulfhydryl oxidases

    PubMed Central

    Heckler, Erin J.; Rancy, Pumtiwitt C.; Kodali, Vamsi K.; Thorpe, Colin

    2008-01-01

    The Quiescin-sulfhydryl oxidase (QSOX) family of flavoenzymes catalyzes the direct and facile insertion of disulfide bonds into unfolded reduced proteins with concomitant reduction of oxygen to hydrogen peroxide. This review discusses the chemical mechanism of these enzymes and the involvement of thioredoxin and flavin-binding domains in catalysis. The variability of CxxC motifs in the QSOX family is highlighted and attention is drawn to the steric factors that may promote efficient thiol/disulfide exchange during oxidative protein folding. The varied cellular location of these multi-domain sulfhydryl oxidases is reviewed and potential intracellular and extracellular roles are summarized. Finally, this review identifies important unresolved questions concerning this ancient family of sulfhydryl oxidases. PMID:17980160

  13. Aerosol optical depth (AOD) and Angstrom exponent of aerosols observed by the Chinese Sun Hazemeter Network from August 2004 to September 2005

    Treesearch

    Jinyuan Xin; Yuesi Wang; Zhanqing Li; Pucai Wang; Wei Min Hao; Bryce L. Nordgren; Shigong Wang; Guangren Lui; Lili Wang; Tianxue Wen; Yang Sun; Bo Hu

    2007-01-01

    To reduce uncertainties in the quantitative assessment of aerosol effects on regional climate and environmental changes, extensive measurements of aerosol optical properties were made with handheld Sun photometers in the Chinese Sun Hazemeter Network (CSHNET) starting in August 2004. Regional characteristics of the aerosol optical depth (AOD) at 500 nm and Angstrom...

  14. THE PREPARATION AND PROPERTIES OF HIGHLY PURIFIED ASCORBIC ACID OXIDASE

    PubMed Central

    Powers, Wendell H.; Lewis, Stanley; Dawson, Charles R.

    1944-01-01

    1. A method is described for the preparation of a highly purified ascorbic acid oxidase containing 0.24 per cent copper. 2. Using comparable activity measurements, this oxidase is about one and a half times as active on a dry weight basis as the hitherto most highly purified preparation described by Lovett-Janison and Nelson. The latter contained 0.15 per cent copper. 3. The oxidase activity is proportional to the copper content and the proportionality factor is the same as that reported by Lovett-Janison and Nelson. 4. When dialyzed free of salt, the blue concentrated oxidase solutions precipitate a dark green-blue protein which carries the activity. This may be prevented by keeping the concentrated solutions about 0.1 M in Na2HPO4. 5. When highly diluted for activity measurements the oxidase rapidly loses activity (irreversibly) previous to the measurement, unless the dilution is made with a dilute inert protein (gelatin) solution. Therefore activity values obtained using such gelatin-stabilized dilute solutions of the oxidase run considerably higher than values obtained by the Lovett-Janison and Nelson technique. 6. The effect of pH and substrate concentration on the activity of the purified oxidase in the presence and absence of inert protein was studied. PMID:19873382

  15. Amine oxidases as important agents of pathological processes of rhabdomyolysis in rats.

    PubMed

    Gudkova, O O; Latyshko, N V; Shandrenko, S G

    2016-01-01

    In this study we have tested an idea on the important role of amine oxidases (semicarbazide-sensitive amine oxidase, diamine oxidase, polyamine oxidase) as an additional source of oxidative/carbonyl stress under glycerol-induced rhabdomyolysis, since the enhanced formation of reactive oxygen species and reactive carbonyl species in a variety of tissues is linked to various diseases. In our experiments we used the sensitive fluorescent method devised for estimation of amine oxidases activity in the rat kidney and thymus as targeted organs under rhabdomyolysis. We have found in vivo the multiple rises in activity of semicarbazide-sensitive amine oxidase, diamine oxidase, polyamine oxidase (2-4.5 times) in the corresponding cell fractions, whole cells or their lysates at the 3-6th day after glycerol injection. Aberrant antioxidant activities depended on rhabdomyolysis stage and had organ specificity. Additional treatment of animals with metal chelator ‘Unithiol’ adjusted only the activity of antioxidant enzymes but not amine oxidases in both organs. Furthermore the in vitro experiment showed that Fenton reaction (hydrogen peroxide in the presence of iron) products alone had no effect on semicarbazide-sensitive amine oxidase activity in rat liver cell fraction whereas supplementation with methylglyoxal resulted in its significant 2.5-fold enhancement. Combined action of the both agents had additive effect on semicarbazide-sensitive amine oxidase activity. We can assume that biogenic amine and polyamine catabolism by amine oxidases is upregulated by oxidative and carbonyl stress factors directly under rhabdomyolysis progression, and the increase in catabolic products concentration contributes to tissue damage in glycerol-induced acute renal failure and apoptosis stimulation in thymus.

  16. Lignin and veratryl alcohol are not inducers of the ligninolytic system of Phanerochaete chrysosporium.

    PubMed Central

    Cancel, A M; Orth, A B; Tien, M

    1993-01-01

    Phanerochaete chrysosporium is a white rot fungus which secretes a family of lignin-degrading enzymes under nutrient limitation. In this work, we investigated the roles of veratryl alcohol and lignin in the ligninolytic system of P. chrysosporium BKM-F-1767 cultures grown under nitrogen-limited conditions. Cultures supplemented with 0.4 to 2 mM veratryl alcohol showed increased lignin peroxidase activity. Addition of veratryl alcohol had no effect on Mn-dependent peroxidase activity and inhibited glyoxal oxidase activity. Azure-casein analysis of acidic proteases in the extracellular fluid showed that protease activity decreased during the early stages of secondary metabolism while lignin peroxidase activity was at its peak, suggesting that proteolysis was not involved in the regulation of lignin peroxidase activity during early secondary metabolism. In cultures supplemented with lignin or veratryl alcohol, no induction of mRNA coding for lignin peroxidase H2 or H8 was observed. Veratryl alcohol protected lignin peroxidase isozymes H2 and H8 from inactivation by H2O2. We conclude that veratryl alcohol acts as a stabilizer of lignin peroxidase activity and not as an inducer of lignin peroxidase synthesis. Images PMID:8215363

  17. Dexamethasone but not indomethacin inhibits human phagocyte nicotinamide adenine dinucleotide phosphate oxidase activity by down-regulating expression of genes encoding oxidase components.

    PubMed

    Condino-Neto, A; Whitney, C; Newburger, P E

    1998-11-01

    We investigated the effects of dexamethasone or indomethacin on the NADPH oxidase activity, cytochrome b558 content, and expression of genes encoding the components gp91-phox and p47-phox of the NADPH oxidase system in the human monocytic THP-1 cell line, differentiated with IFN-gamma and TNF-alpha, alone or in combination, for up to 7 days. IFN-gamma and TNF-alpha, alone or in combination, caused a significant up-regulation of the NADPH oxidase system as reflected by an enhancement of the PMA-stimulated superoxide release, cytochrome b558 content, and expression of gp91-phox and p47-phox genes on both days 2 and 7 of cell culture. Noteworthy was the tremendous synergism between IFN-gamma and TNF-alpha for all studied parameters. Dexamethasone down-regulated the NADPH oxidase system of cytokine-differentiated THP-1 cells as assessed by an inhibition on the PMA-stimulated superoxide release, cytochrome b558 content, and expression of the gp91-phox and p47-phox genes. The nuclear run-on assays indicated that dexamethasone down-regulated the NADPH oxidase system at least in part by inhibiting the transcription of gp91-phox and p47-phox genes. Indomethacin inhibited only the PMA-stimulated superoxide release of THP-1 cells differentiated with IFN-gamma and TNF-alpha during 7 days. None of the other parameters was affected by indomethacin. We conclude that dexamethasone down-regulates the NADPH oxidase system at least in part by inhibiting the expression of genes encoding the gp91-phox and p47-phox components of the NADPH oxidase system.

  18. Heterologous expression and characterization of mouse spermine oxidase.

    PubMed

    Cervelli, Manuela; Polticelli, Fabio; Federico, Rodolfo; Mariottini, Paolo

    2003-02-14

    Polyamine oxidases are key enzymes responsible of the polyamine interconversion metabolism in animal cells. Recently, a novel enzyme belonging to this class of enzymes has been characterized for its capability to oxidize preferentially spermine and designated as spermine oxidase. This is a flavin adenine dinucleotide-containing enzyme, and it has been expressed both in vitro and in vivo systems. The primary structure of mouse spermine oxidase (mSMO) was deduced from a cDNA clone (Image Clone 264769) recovered by a data base search utilizing the human counterpart of polyamine oxidases, PAOh1. The open reading frame predicts a 555-amino acid protein with a calculated M(r) of 61,852.30, which shows a 95.1% identity with PAOh1. To understand the biochemical properties of mSMO and its structure/function relationship, the mSMO cDNA has been subcloned and expressed in secreted and secreted-tagged forms into Escherichia coli BL21 DE3 cells. The recombinant enzyme shows an optimal pH value of 8.0 and is able to oxidize rapidly spermine to spermidine and 3-aminopropanal and fails to act upon spermidine and N(1)-acetylpolyamines. The purified recombinant-tagged form enzyme (M(r) approximately 68,000) has K(m) and k(cat) values of 90 microm and 4.5 s(-1), respectively, using spermine as substrate at pH 8.0. Molecular modeling of mSMO protein based on maize polyamine oxidase three-dimensional structure suggests that the general features of maize polyamine oxidase active site are conserved in mSMO.

  19. Estimating daily PM2.5 and PM10 across the complex geo-climate region of Israel using MAIAC satellite-based AOD data.

    PubMed

    Kloog, Itai; Sorek-Hamer, Meytar; Lyapustin, Alexei; Coull, Brent; Wang, Yujie; Just, Allan C; Schwartz, Joel; Broday, David M

    2015-12-01

    Estimates of exposure to PM 2.5 are often derived from geographic characteristics based on land-use regression or from a limited number of fixed ground monitors. Remote sensing advances have integrated these approaches with satellite-based measures of aerosol optical depth (AOD), which is spatially and temporally resolved, allowing greater coverage for PM 2.5 estimations. Israel is situated in a complex geo-climatic region with contrasting geographic and weather patterns, including both dark and bright surfaces within a relatively small area. Our goal was to examine the use of MODIS-based MAIAC data in Israel, and to explore the reliability of predicted PM 2.5 and PM 10 at a high spatiotemporal resolution. We applied a three stage process, including a daily calibration method based on a mixed effects model, to predict ground PM 2.5 and PM 10 over Israel. We later constructed daily predictions across Israel for 2003-2013 using spatial and temporal smoothing, to estimate AOD when satellite data were missing. Good model performance was achieved, with out-of-sample cross validation R 2 values of 0.79 and 0.72 for PM 10 and PM 2.5 , respectively. Model predictions had little bias, with cross-validated slopes (predicted vs. observed) of 0.99 for both the PM 2.5 and PM 10 models. To our knowledge, this is the first study that utilizes high resolution 1km MAIAC AOD retrievals for PM prediction while accounting for geo-climate complexities, such as experienced in Israel. This novel model allowed the reconstruction of long- and short-term spatially resolved exposure to PM 2.5 and PM 10 in Israel, which could be used in the future for epidemiological studies.

  20. Estimating daily PM2.5 and PM10 across the complex geo-climate region of Israel using MAIAC satellite-based AOD data

    PubMed Central

    Kloog, Itai; Sorek-Hamer, Meytar; Lyapustin, Alexei; Coull, Brent; Wang, Yujie; Just, Allan C.; Schwartz, Joel; Broday, David M.

    2017-01-01

    Estimates of exposure to PM2.5 are often derived from geographic characteristics based on land-use regression or from a limited number of fixed ground monitors. Remote sensing advances have integrated these approaches with satellite-based measures of aerosol optical depth (AOD), which is spatially and temporally resolved, allowing greater coverage for PM2.5 estimations. Israel is situated in a complex geo-climatic region with contrasting geographic and weather patterns, including both dark and bright surfaces within a relatively small area. Our goal was to examine the use of MODIS-based MAIAC data in Israel, and to explore the reliability of predicted PM2.5 and PM10 at a high spatiotemporal resolution. We applied a three stage process, including a daily calibration method based on a mixed effects model, to predict ground PM2.5 and PM10 over Israel. We later constructed daily predictions across Israel for 2003–2013 using spatial and temporal smoothing, to estimate AOD when satellite data were missing. Good model performance was achieved, with out-of-sample cross validation R2 values of 0.79 and 0.72 for PM10 and PM2.5, respectively. Model predictions had little bias, with cross-validated slopes (predicted vs. observed) of 0.99 for both the PM2.5 and PM10 models. To our knowledge, this is the first study that utilizes high resolution 1km MAIAC AOD retrievals for PM prediction while accounting for geo-climate complexities, such as experienced in Israel. This novel model allowed the reconstruction of long- and short-term spatially resolved exposure to PM2.5 and PM10 in Israel, which could be used in the future for epidemiological studies. PMID:28966551

  1. Estimating Ground-Level PM(sub 2.5) Concentrations in the Southeastern United States Using MAIAC AOD Retrievals and a Two-Stage Model

    NASA Technical Reports Server (NTRS)

    Hu, Xuefei; Waller, Lance A.; Lyapustin, Alexei; Wang, Yujie; Al-Hamdan, Mohammad Z.; Crosson, William L.; Estes, Maurice G., Jr.; Estes, Sue M.; Quattrochi, Dale A.; Puttaswamy, Sweta Jinnagara; hide

    2013-01-01

    Previous studies showed that fine particulate matter (PM(sub 2.5), particles smaller than 2.5 micrometers in aerodynamic diameter) is associated with various health outcomes. Ground in situ measurements of PM(sub 2.5) concentrations are considered to be the gold standard, but are time-consuming and costly. Satellite-retrieved aerosol optical depth (AOD) products have the potential to supplement the ground monitoring networks to provide spatiotemporally-resolved PM(sub 2.5) exposure estimates. However, the coarse resolutions (e.g., 10 km) of the satellite AOD products used in previous studies make it very difficult to estimate urban-scale PM(sub 2.5) characteristics that are crucial to population-based PM(sub 2.5) health effects research. In this paper, a new aerosol product with 1 km spatial resolution derived by the Multi-Angle Implementation of Atmospheric Correction (MAIAC) algorithm was examined using a two-stage spatial statistical model with meteorological fields (e.g., wind speed) and land use parameters (e.g., forest cover, road length, elevation, and point emissions) as ancillary variables to estimate daily mean PM(sub 2.5) concentrations. The study area is the southeastern U.S., and data for 2003 were collected from various sources. A cross validation approach was implemented for model validation. We obtained R(sup 2) of 0.83, mean prediction error (MPE) of 1.89 micrograms/cu m, and square root of the mean squared prediction errors (RMSPE) of 2.73 micrograms/cu m in model fitting, and R(sup 2) of 0.67, MPE of 2.54 micrograms/cu m, and RMSPE of 3.88 micrograms/cu m in cross validation. Both model fitting and cross validation indicate a good fit between the dependent variable and predictor variables. The results showed that 1 km spatial resolution MAIAC AOD can be used to estimate PM(sub 2.5) concentrations.

  2. Putting together a plasma membrane NADH oxidase: a tale of three laboratories.

    PubMed

    Löw, Hans; Crane, Frederick L; Morré, D James

    2012-11-01

    The observation that high cellular concentrations of NADH were associated with low adenylate cyclase activity led to a search for the mechanism of the effect. Since cyclase is in the plasma membrane, we considered the membrane might have a site for NADH action, and that NADH might be oxidized at that site. A test for NADH oxidase showed very low activity, which could be increased by adding growth factors. The plasma membrane oxidase was not inhibited by inhibitors of mitochondrial NADH oxidase such as cyanide, rotenone or antimycin. Stimulation of the plasma membrane oxidase by iso-proterenol or triiodothyronine was different from lack of stimulation in endoplasmic reticulum. After 25 years of research, three components of a trans membrane NADH oxidase have been discovered. Flavoprotein NADH coenzyme Q reductases (NADH cytochrome b reductase) on the inside, coenzyme Q in the middle, and a coenzyme Q oxidase on the outside as a terminal oxidase. The external oxidase segment is a copper protein with unique properties in timekeeping, protein disulfide isomerase and endogenous NADH oxidase activity, which affords a mechanism for control of cell growth by the overall NADH oxidase and the remarkable inhibition of oxidase activity and growth of cancer cells by a wide range of anti-tumor drugs. A second trans plasma membrane electron transport system has been found in voltage dependent anion channel (VDAC), which has NADH ferricyanide reductase activity. This activity must be considered in relation to ferricyanide stimulation of growth and increased VDAC antibodies in patients with autism. Copyright © 2012 Elsevier Ltd. All rights reserved.

  3. In Situ Enzymatically Generated Photoswitchable Oxidase Mimetics and Their Application for Colorimetric Detection of Glucose Oxidase.

    PubMed

    Cao, Gen-Xia; Wu, Xiu-Ming; Dong, Yu-Ming; Li, Zai-Jun; Wang, Guang-Li

    2016-07-09

    In this study, a simple and amplified colorimetric assay is developed for the detection of the enzymatic activity of glucose oxidase (GOx) based on in situ formation of a photoswitchable oxidase mimetic of PO₄(3-)-capped CdS quantum dots (QDs). GOx catalyzes the oxidation of 1-thio-β-d-glucose to give 1-thio-β-d-gluconic acid which spontaneously hydrolyzes to β-d-gluconic acid and H₂S; the generated H₂S instantly reacts with Cd(2+) in the presence of Na₃PO₄ to give PO₄(3-)-stabilized CdS QDs in situ. Under visible-light (λ ≥ 400 nm) stimulation, the PO₄(3-)-capped CdS QDs are a new style of oxidase mimic derived by producing some active species, such as h⁺, (•)OH, O₂(•-) and a little H₂O₂, which can oxidize the typical substrate (3,3,5,5-tetramethylbenzydine (TMB)) with a color change. Based on the GOx-triggered growth of the oxidase mimetics of PO₄(3-)-capped CdS QDs in situ, we developed a simple and amplified colorimetric assay to probe the enzymatic activity of GOx. The proposed method allowed the detection of the enzymatic activity of GOx over the range from 25 μg/L to 50 mg/L with a low detection limit of 6.6 μg/L. We believe the PO₄(3-)-capped CdS QDs generated in situ with photo-stimulated enzyme-mimicking activity may find wide potential applications in biosensors.

  4. Genetic and environmental influences on the development of alcoholism: resilience vs. risk.

    PubMed

    Enoch, Mary-Anne

    2006-12-01

    The physiological changes of adolescence may promote risk-taking behaviors, including binge drinking. Approximately 40% of alcoholics were already drinking heavily in late adolescence. Most cases of alcoholism are established by the age of 30 years with the peak prevalence at 18-23 years of age. Therefore the key time frame for the development, and prevention, of alcoholism lies in adolescence and young adulthood. Severe childhood stressors have been associated with increased vulnerability to addiction, however, not all stress-exposed children go on to develop alcoholism. Origins of resilience can be both genetic (variation in alcohol-metabolizing genes, increased susceptibility to alcohol's sedative effects) and environmental (lack of alcohol availability, positive peer and parental support). Genetic vulnerability is likely to be conferred by multiple genes of small to modest effects, possibly only apparent in gene-environment interactions. For example, it has been shown that childhood maltreatment interacts with a monoamine oxidase A (MAOA) gene variant to predict antisocial behavior that is often associated with alcoholism, and an interaction between early life stress and a serotonin transporter promoter variant predicts alcohol abuse in nonhuman primates and depression in humans. In addition, a common Met158 variant in the catechol-O-methyltransferase (COMT) gene can confer both risk and resilience to alcoholism in different drinking environments. It is likely that a complex mix of gene(s)-environment(s) interactions underlie addiction vulnerability and development. Risk-resilience factors can best be determined in longitudinal studies, preferably starting during pregnancy. This kind of research is important for planning future measures to prevent harmful drinking in adolescence.

  5. Assessment of the cerebellar neurotoxic effects of nicotine in prenatal alcohol exposure in rats.

    PubMed

    Bhattacharya, Dwipayan; Majrashi, Mohammed; Ramesh, Sindhu; Govindarajulu, Manoj; Bloemer, Jenna; Fujihashi, Ayaka; Crump, Bailee-Ryan; Hightower, Harrison; Bhattacharya, Subhrajit; Moore, Timothy; Suppiramaniam, Vishnu; Dhanasekaran, Muralikrishnan

    2018-02-01

    The adverse effects of prenatal nicotine and alcohol exposure on human reproductive outcomes are a major scientific and public health concern. In the United States, substantial percentage of women (20-25%) of childbearing age currently smoke cigarettes and consume alcohol, and only a small percentage of these individuals quit after learning of their pregnancy. However, there are very few scientific reports on the effect of nicotine in prenatal alcohol exposure on the cerebellum of the offspring. Therefore, this study was conducted to investigate the cerebellar neurotoxic effects of nicotine in a rodent model of Fetal Alcohol Spectrum Disorder (FASD). In this study, we evaluated the behavioral changes, biochemical markers of oxidative stress and apoptosis, mitochondrial functions and the molecular mechanisms associated with nicotine in prenatal alcohol exposure on the cerebellum. Prenatal nicotine and alcohol exposure induced oxidative stress, did not affect the mitochondrial functions, increased the monoamine oxidase activity, increased caspase expression and decreased ILK, PSD-95 and GLUR1 expression without affecting the GSK-3β. Thus, our current study of prenatal alcohol and nicotine exposure on cerebellar neurotoxicity may lead to new scientific perceptions and novel and suitable therapeutic actions in the future. Copyright © 2017. Published by Elsevier Inc.

  6. R1, a novel repressor of the human monoamine oxidase A.

    PubMed

    Chen, Kevin; Ou, Xiao-Ming; Chen, Gao; Choi, Si Ho; Shih, Jean C

    2005-03-25

    Monoamine oxidase catalyzes the oxidative deamination of a number of neurotransmitters. A deficiency in monoamine oxidase A results in aggressive behavior in both humans and mice. Studies on the regulation of monoamine oxidase A gene expression have shown that the Sp1 family is important for monoamine oxidase A expression. To search for novel transcription factors, the sequences of three Sp1 sites in the monoamine oxidase A core promoter were used in the yeast one-hybrid system to screen a human cDNA library. A novel repressor, R1 (RAM2), has been cloned. The R1 cDNA encodes a protein with 454 amino acids and an open reading frame at the 5'-end. The transfection of R1 in a human neuroblastoma cell line, SK-N-BE (2)-C, inhibited the monoamine oxidase A promoter and enzymatic activity. The degree of inhibition of monoamine oxidase A by R1 correlated with the level of R1 protein expression. R1 was also found to repress monoamine oxidase A promoter activity within a natural chromatin environment. A gel-shift assay indicated that the endogenous R1 protein in SK-N-BE (2)-C cells interacted with the R1 binding sequence. R1 also bound directly to the natural monoamine oxidase A promoter in vivo as shown by chromatin immunoprecipitation assay. Immunocytochemical analysis showed that R1 was expressed in both cytosol and nucleus, which suggested a role for R1 in transcriptional regulation. Northern blot analysis revealed the presence of endogenous R1 mRNA in human brain and peripheral tissues. Taken together, this study shows that R1 is a novel repressor that inhibits monoamine oxidase A gene expression.

  7. A novel proteolytic processing of prolysyl oxidase

    PubMed Central

    Atsawasuwan, Phimon; Mochida, Yoshiyuki; Katafuchi, Michitsuna; Tokutomi, Kentaro; Mocanu, Viorel; Parker, Carol E.; Yamauchi, Mitsuo

    2012-01-01

    Lysyl oxidase (LOX) is an amine oxidase that is critical for the stability of connective tissues. The secreted proLOX is enzymatically quiescent and is activated through proteolytic cleavage between residue Gly162 and Asp163 (residue numbers according to the mouse LOX) by bone morphogenetic protein (BMP)-1 gene products. Here we report a novel processing of proLOX identified in vitro and in vivo. Two forms of mature LOX were identified and characterized by their immunoreactivity to specific antibodies, amine oxidase activity and mass spectrometry. One form was identified as a well characterized BMP-1 processed LOX protein. Another was found to be a truncated form of LOX (tLOX) resulting from the cleavage at the carboxy terminus of Arg192. The tLOX still appeared to retain amine oxidase activity. The results from the proLOX gene deletion and mutation experiments indicated that the processing occurs independent of the cleavage of proLOX by BMP-1 gene products and likely requires the presence of LOX propeptide. These results indicate that proLOX could be processed by two different mechanisms producing two forms of active LOX. PMID:21591931

  8. A novel proteolytic processing of prolysyl oxidase.

    PubMed

    Atsawasuwan, Phimon; Mochida, Yoshiyuki; Katafuchi, Michitsuna; Tokutomi, Kentaro; Mocanu, Viorel; Parker, Carol E; Yamauchi, Mitsuo

    2011-01-01

    Lysyl oxidase (LOX) is an amine oxidase that is critical for the stability of connective tissues. The secreted proLOX is enzymatically quiescent and is activated through proteolytic cleavage between residues Gly(162) and Asp(163) (residue numbers according to the mouse LOX) by bone morphogenetic protein (BMP)-1 gene products. Here we report a novel processing of proLOX identified in vitro and in vivo. Two forms of mature LOX were identified and characterized by their immunoreactivity to specific antibodies, amine oxidase activity, and mass spectrometry. One form was identified as a well-characterized BMP-1 processed LOX protein. Another was found to be a truncated form of LOX resulting from the cleavage at the carboxy terminus of Arg(192). The truncated form of LOX still appeared to retain amine oxidase activity. The results from the proLOX gene deletion and mutation experiments indicated that the processing occurs independent of the cleavage of proLOX by BMP-1 gene products and likely requires the presence of LOX propeptide. These results indicate that proLOX could be processed by two different mechanisms producing two forms of active LOX.

  9. Exploiting algal NADPH oxidase for biophotovoltaic energy

    DOE PAGES

    Anderson, Alexander; Laohavisit, Anuphon; Blaby, Ian K.; ...

    2015-01-29

    Photosynthetic microbes exhibit light-dependent electron export across the cell membrane, which can generate electricity in biological photovoltaic (BPV) devices. How electrons are exported remains to be determined; the identification of mechanisms would help selection or generation of photosynthetic microbes capable of enhanced electrical output. We show that plasma membrane NADPH oxidase activity is a significant component of light-dependent generation of electricity by the unicellular green alga Chlamydomonas reinhardtii. NADPH oxidases export electrons across the plasma membrane to form superoxide anion from oxygen. The C. reinhardtii mutant lacking the NADPH oxidase encoded by RBO1 is impaired in both extracellular superoxide anionmore » production and current generation in a BPV device. Complementation with the wild-type gene restores both capacities, demonstrating the role of the enzyme in electron export. Monitoring light-dependent extracellular superoxide production with a colorimetric assay is shown to be an effective way of screening for electrogenic potential of candidate algal strains. Furthermore, the results show that algal NADPH oxidases are important for superoxide anion production and open avenues for optimizing the biological component of these devices.« less

  10. NADPH oxidases: novel therapeutic targets for neurodegenerative diseases.

    PubMed

    Gao, Hui-Ming; Zhou, Hui; Hong, Jau-Shyong

    2012-06-01

    Oxidative stress is a key pathologic factor in neurodegenerative diseases such as Alzheimer and Parkinson diseases (AD, PD). The failure of free-radical-scavenging antioxidants in clinical trials pinpoints an urgent need to identify and to block major sources of oxidative stress in neurodegenerative diseases. As a major superoxide-producing enzyme complex in activated phagocytes, phagocyte NADPH oxidase (PHOX) is essential for host defense. However, recent preclinical evidence has underscored a pivotal role of overactivated PHOX in chronic neuroinflammation and progressive neurodegeneration. Deficiency in PHOX subunits mitigates neuronal damage induced by diverse insults/stresses relevant to neurodegenerative diseases. More importantly, suppression of PHOX activity correlates with reduced neuronal impairment in models of neurodegenerative diseases. The discovery of PHOX and non-phagocyte NADPH oxidases in astroglia and neurons further reinforces the crucial role of NADPH oxidases in oxidative stress-mediated chronic neurodegeneration. Thus, proper modulation of NADPH oxidase activity might hold therapeutic potential for currently incurable neurodegenerative diseases. Published by Elsevier Ltd.

  11. Dietary Fisetin Supplementation Protects Against Alcohol-Induced Liver Injury in Mice.

    PubMed

    Sun, Qian; Zhang, Wenliang; Zhong, Wei; Sun, Xinguo; Zhou, Zhanxiang

    2016-10-01

    Overproduction of reactive oxygen species is associated with the development of alcoholic liver disease (ALD). Plant polyphenols have been used as dietary interventions for multiple diseases including ALD. The objective of this study was to determine whether dietary supplementation with fisetin, a novel flavonoid, exerts beneficial effect on alcohol-induced liver injury. C57BL/6J mice were pair-fed with the Lieber-DeCarli control or ethanol (EtOH) diet for 4 weeks with or without fisetin supplementation at 10 mg/kg/d. Alcohol feeding induced lipid accumulation in the liver and increased plasma alanine aminotransferase and aspartate aminotransferase activities, which were attenuated by fisetin supplementation. The EtOH concentrations in the plasma and liver were significantly elevated by alcohol exposure but were reduced by fisetin supplementation. Although fisetin did not affect the protein expression of alcohol metabolism enzymes, the aldehyde dehydrogenase activities were significantly increased by fisetin compared to the alcohol alone group. In addition, fisetin supplementation remarkably reduced hepatic NADPH oxidase 4 levels along with decreased plasma hydrogen peroxide and hepatic superoxide and 4-hydroxynonenal levels after alcohol exposure. Alcohol-induced apoptosis and up-regulation of Fas and cleaved caspase-3 in the liver were prevented by fisetin. Moreover, fisetin supplementation attenuated alcohol-induced hepatic steatosis through increasing plasma adiponectin levels and hepatic protein levels of p-AMPK, ACOX1, CYP4A, and MTTP. This study demonstrated that the protective effect of fisetin on ALD is achieved by accelerating EtOH clearance and inhibition of oxidative stress. The data suggest that fisetin has a therapeutical potential for treating ALD. Copyright © 2016 by the Research Society on Alcoholism.

  12. Pulse-radiolysis studies on the interaction of one-electron reduced species with blue oxidases. Reduction of type-2-copper-depleted ascorbate oxidase.

    PubMed

    O'Neill, P; Fielden, E M; Avigliano, L; Marcozzi, G; Ballini, A; Agrò, F

    1984-08-15

    The interaction of one-electron reduced metronidazole (ArNO2.-) with native and Type-2-copper-depleted ascorbate oxidase were studied in buffered aqueous solution at pH 6.0 and 7.4 by using the technique of pulse radiolysis. With ArNO2.-, reduction of Type 1 copper of the native enzyme and of the Type-2-copper-depleted ascorbate oxidase occurs via a bimolecular step and at the same rate. Whereas the native protein accepts, in the absence of O2, 6-7 reducing equivalents, Type-2-copper-depleted ascorbate oxidase accepts only 3 reducing equivalents with stoichiometric reduction of Type 1 copper. On reaction of O2.- with ascorbate oxidase under conditions of [O2.-] much greater than [ascorbate oxidase], removal of Type 2 copper results in reduction of all the Type 1 copper atoms, in contrast with reduction of the equivalent of only one Type 1 copper atom in the holoprotein. From observations at 610 nm, the rate of reduction of ascorbate oxidase by O2.- is not dependent on the presence of Type 2 copper. For the holoprotein, no significant optical-absorption changes were observed at 330 nm. It is proposed that electrons enter the protein via Type 1 copper in a rate-determining step followed by a fast intramolecular transfer of electrons within the protein. For the Type-2-copper-depleted protein, intramolecular transfer within the protein, however, is slow or does not occur. In the presence of O2, it is also suggested that re-oxidation of the partially reduced holoprotein occurs at steady state, as inferred from the observations at 330 nm and 610 nm. The role of Type 2 copper in ascorbate oxidase is discussed in terms of its involvement in redistribution of electrons within the protein or structural considerations.

  13. Supramolecular organization of cytochrome c oxidase- and alternative oxidase-dependent respiratory chains in the filamentous fungus Podospora anserina.

    PubMed

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

    2004-06-18

    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.

  14. In vitro study of 6-mercaptopurine oxidation catalysed by aldehyde oxidase and xanthine oxidase.

    PubMed

    Rashidi, Mohammad-Reza; Beedham, Christine; Smith, John S; Davaran, Soodabeh

    2007-08-01

    In spite of over 40 years of clinical use of 6-mercaptopurine, many aspects of complex pharmacology and metabolism of this drug remain unclear. It is thought that 6-mercaptopurine is oxidized to 6-thiouric acid through 6-thioxanthine or 8-oxo-6-mercaptopurine by one of two molybdenum hydroxylases, xanthine oxidase (XO), however, the role of other molybdenum hydroxylase, aldehyde oxidase (AO), in the oxidation of 6-mercaptopurine and possible interactions of AO substrates and inhibitors has not been investigated in more details. In the present study, the role of AO and XO in the oxidation of 6- mercaptopurine has been investigated. 6-mercaptopurine was incubated with bovine milk xanthine oxidase or partially purified guinea pig liver molybdenum hydroxylase fractions in the absence and presence of XO and AO inhibitor/substrates, and the reactions were monitored by spectrophotometric and HPLC methods. According to the results obtained from the inhibition studies, it is more likely that 6- mercaptopurine is oxidized to 6-thiouric acid via 6-thioxanthine rather than 8-oxo-6-mercaptopurine. The first step which is the rate limiting step is catalyzed solely by XO, whereas both XO and AO are involved in the oxidation of 6-thioxanthine to 6-thiouric acid.

  15. Analysis of CO, CH4 and AOD distributions over Eurasia and estimates of their long-term tendencies based on spectroscopic ground-based and satellite observations

    NASA Astrophysics Data System (ADS)

    Rakitin, Vadim; Elansky, Nikolai; Shtabkin, Yury; Dzhola, Anatoly; Pankratova, Natalia; Shilkin, Arseny

    2017-04-01

    Analysis of the CO and CH4 total column (TC) measurements and AOD data in urban and background regions of Eurasia for period from 1998 to 2016 years is presented. The trends estimates based on spectroscopic ground-based datasets of OIAP, SPSU, IAP CAC, NPO "Typhoon" and NDACC were compared with similar ones obtained with use of orbital data (MOPITT v6J and AIRS v6). Total decrease of CO TC in both urban (Moscow and Beijing) and background regions (ZSS, Peterhof, Obninsk, European NDACC sites) in 1998-2016 years changed to increase of CO in summer and autumn months in almost all background regions of Northern Eurasia after 2007. Negative trends of AOD were obtained for Europe, West Siberia and China for different seasons (including summer and autumn months) for time periods 2000-2016 and 2007-2016 with using both AERONET and MODIS Terra/Aqua datasets; AOD trends over East Siberia were positive that dui to influence of strong wild fires in 2010-2016 years in Siberia. Rate of CO TC decrease obtained with orbital data using are less than the same for ground based data with factor 1.5-2.0 for both urban and background regions. Rate of CH4 TC increased after 2007 in North-West Eurasian regions and didn't change in most of North-East regions. The negative AOD trends over Europe and West Siberia indirectly point to non-increase of wild-fires emissions over this region in latest years. Therefore the positive CO TC trends cannot be explained only by increase of wild-fires impact and anthropogenic emissions; possible reasons of such CO tendencies could be the changes in all atmospheric photochemistry system. This work was supported by the Russian Scientific Foundation under grant №14-47-00049 (in part of NDACC, AERONET and satellite trends estimates), under grant №16-17-10275 (in part of analysis of ground-based observations over Moscow and Obninsk) and partially by the Russian Foundation for Basic Research (grant № 16-05-00287 in part of provide of ground

  16. Stability of spermine oxidase to thermal and chemical denaturation: comparison with bovine serum amine oxidase.

    PubMed

    Cervelli, Manuela; Leonetti, Alessia; Cervoni, Laura; Ohkubo, Shinji; Xhani, Marla; Stano, Pasquale; Federico, Rodolfo; Polticelli, Fabio; Mariottini, Paolo; Agostinelli, Enzo

    2016-10-01

    Spermine oxidase (SMOX) is a flavin-containing enzyme that specifically oxidizes spermine to produce spermidine, 3-aminopropanaldehyde and hydrogen peroxide. While no crystal structure is available for any mammalian SMOX, X-ray crystallography showed that the yeast Fms1 polyamine oxidase has a dimeric structure. Based on this scenario, we have investigated the quaternary structure of the SMOX protein by native gel electrophoresis, which revealed a composite gel band pattern, suggesting the formation of protein complexes. All high-order protein complexes are sensitive to reducing conditions, showing that disulfide bonds were responsible for protein complexes formation. The major gel band other than the SMOX monomer is the covalent SMOX homodimer, which was disassembled by increasing the reducing conditions, while being resistant to other denaturing conditions. Homodimeric and monomeric SMOXs are catalytically active, as revealed after gel staining for enzymatic activity. An engineered SMOX mutant deprived of all but two cysteine residues was prepared and characterized experimentally, resulting in a monomeric species. High-sensitivity differential scanning calorimetry of SMOX was compared with that of bovine serum amine oxidase, to analyse their thermal stability. Furthermore, enzymatic activity assays and fluorescence spectroscopy were used to gain insight into the unfolding process.

  17. POLYAMINE OXIDASE 1 from rice (Oryza sativa) is a functional ortholog of Arabidopsis POLYAMINE OXIDASE 5.

    PubMed

    Liu, Taibo; Wook Kim, Dong; Niitsu, Masaru; Berberich, Thomas; Kusano, Tomonobu

    2014-01-01

    POLYAMINE OXIDASE 1 (OsPAO1), from rice (Oryza sativa), and POLYAMINE OXIDASE 5 (AtPAO5), from Arabidopsis (Arabidopsis thaliana), are enzymes sharing high identity at the amino acid level and with similar characteristics, such as polyamine specificity and pH preference; furthermore, both proteins localize to the cytosol. A loss-of-function Arabidopsis mutant, Atpao5-2, was hypersensitive to low doses of exogenous thermospermine but this phenotype could be rescued by introduction of the wild-type AtPAO5 gene. Introduction of OsPAO1, under the control of a constitutive promoter, into Atpao5-2 mutants also restored normal thermospermine sensitivity, allowing growth in the presence of low levels of thermospermine, along with a concomitant decrease in thermospermine content in plants. By contrast, introduction of OsPAO3, which encodes a peroxisome-localized polyamine oxidase, into Atpao5-2 plants could not rescue any of the mutant phenotypes in the presence of thermospermine. These results suggest that OsPAO1 is the functional ortholog of AtPAO5.

  18. Aurone synthase is a catechol oxidase with hydroxylase activity and provides insights into the mechanism of plant polyphenol oxidases

    PubMed Central

    Molitor, Christian; Mauracher, Stephan Gerhard

    2016-01-01

    Tyrosinases and catechol oxidases belong to the family of polyphenol oxidases (PPOs). Tyrosinases catalyze the o-hydroxylation and oxidation of phenolic compounds, whereas catechol oxidases were so far defined to lack the hydroxylation activity and catalyze solely the oxidation of o-diphenolic compounds. Aurone synthase from Coreopsis grandiflora (AUS1) is a specialized plant PPO involved in the anabolic pathway of aurones. We present, to our knowledge, the first crystal structures of a latent plant PPO, its mature active and inactive form, caused by a sulfation of a copper binding histidine. Analysis of the latent proenzyme’s interface between the shielding C-terminal domain and the main core provides insights into its activation mechanisms. As AUS1 did not accept common tyrosinase substrates (tyrosine and tyramine), the enzyme is classified as a catechol oxidase. However, AUS1 showed hydroxylase activity toward its natural substrate (isoliquiritigenin), revealing that the hydroxylase activity is not correlated with the acceptance of common tyrosinase substrates. Therefore, we propose that the hydroxylase reaction is a general functionality of PPOs. Molecular dynamics simulations of docked substrate–enzyme complexes were performed, and a key residue was identified that influences the plant PPO’s acceptance or rejection of tyramine. Based on the evidenced hydroxylase activity and the interactions of specific residues with the substrates during the molecular dynamics simulations, a novel catalytic reaction mechanism for plant PPOs is proposed. The presented results strongly suggest that the physiological role of plant catechol oxidases were previously underestimated, as they might hydroxylate their—so far unknown—natural substrates in vivo. PMID:26976571

  19. Impact of diurnal variability and meteorological factors on the PM2.5 - AOD relationship: Implications for PM2.5 remote sensing.

    PubMed

    Guo, Jianping; Xia, Feng; Zhang, Yong; Liu, Huan; Li, Jing; Lou, Mengyun; He, Jing; Yan, Yan; Wang, Fu; Min, Min; Zhai, Panmao

    2017-02-01

    PM 2.5 retrieval from space is still challenging due to the elusive relationship between PM 2.5 and aerosol optical depth (AOD), which is further complicated by meteorological factors. In this work, we investigated the diurnal cycle of PM 2.5 in China, using ground-based PM measurements obtained at 226 sites of China Atmosphere Watch Network during the period of January 2013 to December 2015. Results showed that nearly half of the sites witnessed a PM 2.5 maximum in the morning, in contrast to the least frequent occurrence (5%) in the afternoon when strong solar radiation received at the surface results in rapid vertical diffusion of aerosols and thus lower mass concentration. PM 2.5 tends to peak equally in the morning and evening in North China Plain (NCP) with an amplitude of nearly twice or three times that in the Pearl River Delta (PRD), whereas the morning PM 2.5 peak dominates in Yangtze River Delta (YRD) with a magnitude lying between those of NCP and PRD. The gridded correlation maps reveal varying correlations around each PM 2.5 site, depending on the locations and seasons. Concerning the impact of aerosol diurnal variation on the correlation, the averaging schemes of PM 2.5 using 3-h, 5-h, and 24-h time windows tend to have larger R biases, compared with the scheme of 1-h time window, indicating diurnal variation of aerosols plays a significant role in the establishment of explicit correlation between PM 2.5 and AOD. In addition, high cloud fraction and relative humidity tend to weaken the correlation, regardless of geographical location. Therefore, the impact of meteorology could be one of the most plausible alternatives in explaining the varying R values observed, due to its non-negligible effect on MODIS AOD retrievals. Our findings have implications for PM 2.5 remote sensing, as long as the aerosol diurnal cycle, along with meteorology, are explicitly considered in the future. Copyright © 2016 The Authors. Published by Elsevier Ltd.. All rights

  20. Three-dimensional organization of three-domain copper oxidases: A review

    NASA Astrophysics Data System (ADS)

    Zhukhlistova, N. E.; Zhukova, Yu. N.; Lyashenko, A. V.; Zaĭtsev, V. N.; Mikhaĭlov, A. M.

    2008-01-01

    “Blue” copper-containing proteins are multidomain proteins that utilize a unique redox property of copper ions. Among other blue multicopper oxidases, three-domain oxidases belong to the group of proteins that exhibit a wide variety of compositions in amino acid sequences, functions, and occurrences in organisms. This paper presents a review of the data obtained from X-ray diffraction investigations of the three-dimensional structures of three-domain multicopper oxidases, such as the ascorbate oxidase catalyzing oxidation of ascorbate to dehydroascorbate and its three derivatives; the multicopper oxidase CueO (the laccase homologue); the laccases isolated from the basidiomycetes Coprinus cinereus, Trametes versicolor, Coriolus zonatus, Cerrena maxima, and Rigidoporus lignosus and the ascomycete Melanocarpus albomyces; and the bacterial laccases CotA from the endospore coats of Bacillus subtilis. A comparison of the molecular structures of the laccases of different origins demonstrates that, structurally, these objects are highly conservative. This obviously indicates that the catalytic activity of the enzymes under consideration is characterized by similar mechanisms.

  1. NADPH OXIDASE: STRUCTURE AND ACTIVATION MECHANISMS (REVIEW). NOTE I.

    PubMed

    Filip-Ciubotaru, Florina; Manciuc, Carmen; Stoleriu, Gabriela; Foia, Liliana

    2016-01-01

    NADPH oxidase (nicotinamide adenine dinucleotide phosphate-oxidase), with its generically termed NOX isoforms, is the major source of ROS (reactive oxigen species) in biological systems. ROS are small oxygen-derived molecules with an important role in various biological processes (physiological or pathological). If under physiological conditions some processes are beneficial and necessary for life, under pathophysiological conditions they are noxious, harmful. NADPH oxidases are present in phagocytes and in a wide variety of nonphagocytic cells. The enzyme generates superoxide by transferring electrons from NADPH inside the cell across the membrane and coupling them to molecular oxygen to produce superoxide anion, a reactive free-radical. Structurally, NADPH oxidase is a multicomponent enzyme which includes two integral membrane proteins, glycoprotein gp9 1 Phox and adaptor protein p22(phox), which together form the heterodimeric flavocytochrome b558 that constitutes the core of the enzyme. During the resting state, the multidomain regulatory subunits p40P(phox), p47(phox), p67(Phox) are located in the cytosol organized as a complex. The activation of phagocytic NADPH oxidase occurs through a complex series of protein interactions.

  2. Dietary fisetin supplementation protects against alcohol-induced liver injury in mice

    PubMed Central

    Sun, Qian; Zhang, Wenliang; Zhong, Wei; Sun, Xinguo; Zhou, Zhanxiang

    2016-01-01

    Background Overproduction of reactive oxygen species (ROS) is associated with the development of alcoholic liver disease (ALD). Plant polyphenols have been used as dietary inverventions for multiple diseases including ALD. The objective of the present study was to determine whether dietary supplementation with fisetin, a novel flavonoid, exerts beneficial effect on alcohol-induced liver injury. Methods C57BL/6J mice were pair-fed with the Lieber-DeCarli control or ethanol diet for four weeks with or without fisetin supplementation at 10 mg/kg/d. Results Alcohol feeding induced lipid accumulation in the liver and increased plasma ALT and AST activities, which were attenuated by fisetin suplementation. The ethanol concentrations in the plasma and liver were significantly elevated by alcohol exposure but were reduced by fisetin suplementation. Although fisetin did not affect the protein expression of alcohol metabolism enzymes, the aldehyde dehydrogenase activities were significantly increased by fisetin compared to the alcohol alone group. In addition, fisetin suplementation remarkably reduced hepatic NADPH oxidase 4 (NOX4) levels along with decreased plasma hydrogen peroxide and hepatic superoxide and 4-hydroxynonenal (4HNE) levels after alcohol exposure. Alcohol-induced apoptosis and upregulation of Fas and cleaved caspase-3 in the liver were prevented by fisetin. Moreover, fisetin suplementation attenuated alcohol-induced hepatic streatosis through increasing plasma adiponectin levels and hepatic protein levels of p-AMPK, ACOX1, CYP4A, and MTTP. Conclusion The present study demonstrated that the protective effect of fisetin on ALD is achieved by accelerating ethanol clearance and inhibition of oxidative stress. The data suggest that fisetin has a therapeutical potential for treating ALD. PMID:27575873

  3. Expression and Chloroplast Targeting of Cholesterol Oxidase in Transgenic Tobacco Plants

    PubMed Central

    Corbin, David R.; Grebenok, Robert J.; Ohnmeiss, Thomas E.; Greenplate, John T.; Purcell, John P.

    2001-01-01

    Cholesterol oxidase represents a novel type of insecticidal protein with potent activity against the cotton boll weevil (Anthonomus grandis grandis Boheman). We transformed tobacco (Nicotiana tabacum) plants with the cholesterol oxidase choM gene and expressed cytosolic and chloroplast-targeted versions of the ChoM protein. Transgenic leaf tissues expressing cholesterol oxidase exerted insecticidal activity against boll weevil larvae. Our results indicate that cholesterol oxidase can metabolize phytosterols in vivo when produced cytosolically or when targeted to chloroplasts. The transgenic plants exhibiting cytosolic expression accumulated low levels of saturated sterols known as stanols, and displayed severe developmental aberrations. In contrast, the transgenic plants expressing chloroplast-targeted cholesterol oxidase maintained a greater accumulation of stanols, and appeared phenotypically and developmentally normal. These results are discussed within the context of plant sterol distribution and metabolism. PMID:11457962

  4. Substance abuse prevention in Cape Town's peri-urban settlements: local health trainers' perspectives

    PubMed Central

    Puljević, Cheneal; Learmonth, Despina

    2014-01-01

    South Africa currently experiences high levels of alcohol and other drug (AOD) abuse. As a result there is a need for the initiation of regional AOD abuse prevention programmes with a specific focus on youth prevention strategies. The Medical Knowledge Institute (MKI) is a non-profit organisation which develops and facilitates health information workshops to members of disadvantaged peri-urban communities in South Africa. This research investigated the views of eight local MKI health trainers on factors contributing to AOD abuse in their communities. Although the expected focus of the discussion was on prevention strategies and effective interventions, the trainers placed more emphasis on the individual and community factors influencing AOD abuse. The themes which emerged through the research included: status, government, (di)stress, gender, recreation, consequences and community. This research holds significance as it has the potential to assist further development of community-based AOD prevention workshops and to guide public health policy and service development for AOD abuse. PMID:25750776

  5. Comparative Activity-Based Flavin-Dependent Oxidase Profiling.

    PubMed

    Krysiak, Joanna; Breinbauer, Rolf

    2017-01-01

    Activity-based protein profiling (ABPP) has become a powerful chemoproteomic technology allowing for the dissection of complex ligand-protein interactions in their native cellular environment. One of the biggest challenges for ABPP is the extension of the proteome coverage. In this chapter a new ABPP strategy dedicated to monoamine oxidases (MAO) is presented. These enzymes are representative examples of flavin-dependent oxidases, playing a crucial role in the regulation of nervous system signaling.

  6. Multidomain flavin-dependent sulfhydryl oxidases.

    PubMed

    Coppock, Donald L; Thorpe, Colin

    2006-01-01

    Eukaryotic flavin-dependent sulfhydryl oxidases catalyze oxidative protein folding with the generation of disulfides and the reduction of oxygen to hydrogen peroxide. This review deals principally with the Quiescinsulfhydryl oxidases (QSOX) that are found in multiple forms in multicellular organisms and singly in a number of protozoan parasites. QSOX is an ancient fusion of thioredoxin domains and an FAD-binding module, ERV1/ALR. Interdomain disulfide exchanges transmit reducing equivalents from substrates to the flavin cofactor and thence to molecular oxygen. The in vitro substrate specificity of avian QSOX1 and the likely substrates of QSOXs in vivo are discussed. The location of QSOX immunoreactivity and mRNA expression levels in human cells and tissues is reviewed. Generally, there is a marked association of QSOX1 expression with cell types that have a high secretory load of disulfide-containing peptides and proteins. The abundance of sulfhydryl oxidases in the islets of Langerhans suggests that oxidative protein folding may directly contribute to the oxidative stress believed to be a factor in the progression to type II diabetes. Finally, the structure and mechanism of QSOX proteins is compared to their smaller stand-alone cousins: yeast ERV1p and ERV2p, the mammalian augmenter of liver regeneration (ALR), and the viral ALR homologs.

  7. Getting It Together: Promoting Drug-Free Communities. A Resource Guide for Developing Effective Youth Coalitions.

    ERIC Educational Resources Information Center

    Circle, Inc., McLean, VA.

    This guide is designed to serve organizations and individuals working with youth on issues of alcohol and other drug (AOD) prevention. Chapter 1 examines why coalitions are needed, characteristics of prevention, misconceptions about alcohol and other drug use, community systems and alcohol and other drug use, definitions of coalitions and…

  8. Effect of contraceptive steroids on monoamine oxidase activity

    PubMed Central

    Southgate, Jennifer; Collins, G. G. S.; Pryse-Davies, J.; Sandler, M.

    1969-01-01

    Cyclical variations in monoamine oxidase activity during the human menstrual cycle, specific to the endometrium and modified in women undergoing contraceptive steroid treatment, may reflect changes in hormonal environment. Treatment of rats with individual constituents of the contraceptive pill causes analogous changes: oestrogens inhibit and progestogens potentiate uterine monoamine oxidase activity. ImagesFig. 2Fig. 3

  9. Plasma diamine oxidase levels in pregnancy complicated by threatened abortion.

    PubMed Central

    Legge, M; Duff, G B

    1981-01-01

    Plasma diamine oxidase levels were assayed in 66 patients who presented with pregnancy complicated by threatened abortion. Levels within the normal range were associated with continuing pregnancies, whereas levels below the normal range were associated with subsequent abortion. Among those patients in whom gestation was greater than eight weeks, 66.6% of diamine oxidase levels correctly predicted the pregnancy outcome. Assay of the diamine oxidase levels at eight weeks of gestation or less gave little useful information. PMID:6785320

  10. Plasma diamine oxidase levels in pregnancy complicated by threatened abortion.

    PubMed

    Legge, M; Duff, G B

    1981-02-01

    Plasma diamine oxidase levels were assayed in 66 patients who presented with pregnancy complicated by threatened abortion. Levels within the normal range were associated with continuing pregnancies, whereas levels below the normal range were associated with subsequent abortion. Among those patients in whom gestation was greater than eight weeks, 66.6% of diamine oxidase levels correctly predicted the pregnancy outcome. Assay of the diamine oxidase levels at eight weeks of gestation or less gave little useful information.

  11. Expression of Ascorbic Acid Oxidase in Zucchini Squash (Cucurbita pepo L.).

    PubMed

    Lin, L S; Varner, J E

    1991-05-01

    The expression of ascorbic acid oxidase was studied in zucchini squash (Cucurbita pepo L.), one of the most abundant natural sources of the enzyme. In the developing fruit, specific activity of ascorbic acid oxidase was highest between 4 and 6 days after anthesis. Protein and mRNA levels followed the same trend as enzyme activity. Highest growth rate of the fruit occurred before 6 days after anthesis. Within a given fruit, ascorbic acid oxidase activity and mRNA level were highest in the epidermis, and lowest in the central placental region. In leaf tissue, ascorbic acid oxidase activity was higher in young leaves, and very low in old leaves. Within a given leaf, enzyme activity was highest in the fast-growing region (approximately the lower third of the blade), and lowest in the slow-growing region (near leaf apex). High expression of ascorbic acid oxidase at a stage when rapid growth is occurring (in both fruits and leaves), and localization of the enzyme in the fruit epidermis, where cells are under greatest tension during rapid growth in girth, suggest that ascorbic acid oxidase might be involved in reorganization of the cell wall to allow for expansion. Based on the known chemistry of dehydroascorbic acid, the end product of the ascorbic acid oxidase-catalyzed reaction, we have proposed several hypotheses to explain how dehydroascorbic acid might cause cell wall "loosening."

  12. Structure-function relationships in the evolutionary framework of spermine oxidase.

    PubMed

    Cervelli, Manuela; Salvi, Daniele; Polticelli, Fabio; Amendola, Roberto; Mariottini, Paolo

    2013-06-01

    Spermine oxidase is a FAD-dependent enzyme that specifically oxidizes spermine, and plays a central role in the highly regulated catabolism of polyamines in vertebrates. The spermine oxidase substrate is specifically spermine, a tetramine that plays mandatory roles in several cell functions, such as DNA synthesis, cellular proliferation, modulation of ion channels function, cellular signalling, nitric oxide synthesis and inhibition of immune responses. The oxidative products of spermine oxidase activity are spermidine, H2O2 and the aldehyde 3-aminopropanal that spontaneously turns into acrolein. In this study the reconstruction of the phylogenetic relationships among spermine oxidase proteins from different vertebrate taxa allowed to infer their molecular evolutionary history, and assisted in elucidating the conservation of structural and functional properties of this enzyme family. The amino acid residues, which have been hypothesized or demonstrated to play a pivotal role in the enzymatic activity, and substrate specificity are here analysed to obtain a comprehensive and updated view of the structure-function relationships in the evolution of spermine oxidase.

  13. Three-dimensional organization of three-domain copper oxidases: A review

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

    Zhukhlistova, N. E., E-mail: amm@ns.crys.ras.ru; Zhukova, Yu. N.; Lyashenko, A. V.

    2008-01-15

    'Blue' copper-containing proteins are multidomain proteins that utilize a unique redox property of copper ions. Among other blue multicopper oxidases, three-domain oxidases belong to the group of proteins that exhibit a wide variety of compositions in amino acid sequences, functions, and occurrences in organisms. This paper presents a review of the data obtained from X-ray diffraction investigations of the three-dimensional structures of three-domain multicopper oxidases, such as the ascorbate oxidase catalyzing oxidation of ascorbate to dehydroascorbate and its three derivatives; the multicopper oxidase CueO (the laccase homologue); the laccases isolated from the basidiomycetes Coprinus cinereus, Trametes versicolor, Coriolus zonatus, Cerrenamore » maxima, and Rigidoporus lignosus and the ascomycete Melanocarpus albomyces; and the bacterial laccases CotA from the endospore coats of Bacillus subtilis. A comparison of the molecular structures of the laccases of different origins demonstrates that, structurally, these objects are highly conservative. This obviously indicates that the catalytic activity of the enzymes under consideration is characterized by similar mechanisms.« less

  14. Biochemistry of microbial polyvinyl alcohol degradation.

    PubMed

    Kawai, Fusako; Hu, Xiaoping

    2009-08-01

    Effect of minor chemical structures such as 1,2-diol content, ethylene content, tacticity, a degree of polymerization, and a degree of saponification of the main chain on biodegradability of polyvinyl alcohol (PVA) is summarized. Most PVA-degraders are Gram-negative bacteria belonging to the Pseudomonads and Sphingomonads, but Gram-positive bacteria also have PVA-degrading abilities. Several examples show symbiotic degradation of PVA by different mechanisms. Penicillium sp. is the only reported eukaryotic degrader. A vinyl alcohol oligomer-utilizing fungus, Geotrichum fermentans WF9101, has also been reported. Lignolytic fungi have displayed non-specific degradation of PVA. Extensive published studies have established a two-step process for the biodegradation of PVA. Some bacteria excrete extracellular PVA oxidase to yield oxidized PVA, which is partly under spontaneous depolymerization and is further metabolized by the second step enzyme (hydrolase). On the other hand, PVA (whole and depolymerized to some extent) must be taken up into the periplasmic space of some Gram-negative bacteria, where PVA is oxidized by PVA dehydrogenase, coupled to a respiratory chain. The complete pva operon was identified in Sphingopyxis sp. 113P3. Anaerobic biodegradability of PVA has also been suggested.

  15. Polyenylphosphatidylcholine attenuates alcohol-induced fatty liver and hyperlipemia in rats.

    PubMed

    Navder, K P; Baraona, E; Lieber, C S

    1997-09-01

    Chronic administration of a soybean-derived polyenylphosphatidylcholine (PPC) extract prevents the development of cirrhosis in alcohol-fed baboons. To assess whether this phospholipid also affects earlier changes induced by alcohol consumption (such as fatty liver and hyperlipemia), 28 male rat littermates were pair-fed liquid diets containing 36% of energy either as ethanol or as additional carbohydrate for 21 d, and killed 90 min after intragastric administration of the corresponding diets. Half of the rats were given PPC (3 g/l), whereas the other half received the same amount of linoleate (as safflower oil) and choline (as bitartrate salt). PPC did not affect diet or alcohol consumption [15.4 +/- 0.5 G/(kg.d)], but the ethanol-induced hepatomegaly and the hepatic accumulation of lipids (principally triglycerides and cholesterol esters) and proteins were about half those in rats not given PPC. The ethanol-induced postprandial hyperlipemia was lower with PPC than without, despite an enhanced fat absorption and no difference in the level of plasma free fatty acids. The attenuation of fatty liver and hyperlipemia was associated with correction of the ethanol-induced inhibition of mitochondrial oxidation of palmitoyl-1-carnitine and the depression of cytochrome oxidase activity, as well as the increases in activity of serum glutamate dehydrogenase and aminotransferases. Thus, PPC attenuates early manifestations of alcohol toxicity, at least in part, by improving mitochondrial injury. These beneficial effects of PPC at the initial stages of alcoholic liver injury may prevent or delay the progression to more advanced forms of alcoholic liver disease.

  16. Catalase deficiency may complicate urate oxidase (rasburicase) therapy.

    PubMed

    Góth, László; Bigler, N William

    2007-09-01

    Patients with low (inherited and acquired) catalase activities who are treated with infusion of uric acid oxidase because they are at risk of tumour lysis syndrome may experience very high concentrations of hydrogen peroxide. They may suffer from methemoglobinaemia and haemolytic anaemia which may be attributed either to deficiency of glucose-6-phosphate dehydrogenase or to other unknown circumstances. Data have not been reported from catalase deficient patients who were treated with uric acid oxidase. It may be hypothesized that their decreased blood catalase could lead to the increased concentration of hydrogen peroxide which may cause haemolysis and formation of methemoglobin. Blood catalase activity should be measured for patients at risk of tumour lysis syndrome prior to uric acid oxidase treatment.

  17. Characterization of the fine fraction of the argon oxygen decarburization with lance (AOD-L) sludge generated by the stainless steelmaking industry.

    PubMed

    Majuste, Daniel; Mansur, Marcelo Borges

    2008-05-01

    The argon oxygen decarburization with lance (AOD-L) sludge generated by the stainless steelmaking industry is a hazardous waste due to the presence of chromium. While its coarse fraction is usually recycled into the own industrial process, the fine fraction is normally disposed in landfills. Techniques such as briquetting or magnetic separation were found to be inadequate to treat it for reuse purposes. So, in this work, the fine fraction of the AOD-L sludge was characterized aiming to find alternative methods to treat it. This sludge consists of a fine powder (mean diameter of 1 microm) containing 34 +/- 2% (w/w) of iron, 10.2 +/- 0.9% (w/w) of chromium and 1.4 +/- 0.1% (w/w) of nickel. The main crystalline phases identified in this study were chromite (FeCr(2)O(4)), magnetite (Fe(3)O(4)), hematite (Fe(2)O(3)) and calcite (CaCO(3)). In the digestion tests, the addition of HClO(4) has favored the dissolution of chromite which is a very stable oxide in aqueous media. Nickel was found in very fine particles, probably in the metallic form or associated with iron and oxygen. The sludge was classified as hazardous waste, so its disposal in landfills should be avoided.

  18. Aircraft- and ground-based assessment of the CCN-AOD relationship and implications on model analysis of ACI and underlying aerosol processes

    NASA Astrophysics Data System (ADS)

    Shinozuka, Y.; Clarke, A. D.; Nenes, A.; Lathem, T. L.; Redemann, J.; Jefferson, A.; Wood, R.

    2014-12-01

    Contrary to common assumptions in satellite-based modeling of aerosol-cloud interactions, ∂logCCN/∂logAOD is less than unity, i.e., the number concentration of cloud condensation nuclei (CCN) less than doubles as aerosol optical depth (AOD) doubles. This can be explained by omnipresent aerosol processes. Condensation, coagulation and cloud processing, for example, generally make particles scatter more light while hardly increasing their number. This paper reports on the relationship in local air masses between CCN concentration, aerosol size distribution and light extinction observed from aircraft and the ground at diverse locations. The CCN-to-local-extinction relationship, when averaged over ~1 km distance and sorted by the wavelength dependence of extinction, varies approximately by a factor of 2, reflecting the variability in aerosol intensive properties. This, together with retrieval uncertainties and the variability in aerosol spatio-temporal distribution and hygroscopic growth, challenges satellite-based CCN estimates. However, the large differences in estimated CCN may correspond to a considerably lower uncertainty in cloud drop number concentration (CDNC), given the sublinear response of CDNC to CCN. Overall, our findings from airborne and ground-based observations call for model-based reexamination of aerosol-cloud interactions and underlying aerosol processes.

  19. [Respiratory oxidases: the enzymes which use most of the oxygen which living things breathe].

    PubMed

    Toledo-Cuevas, E M

    1997-01-01

    The respiratory oxidases are the last enzymes of the aerobic respiratory chain. They catalize the reduction of molecular oxygen to water, with generation of an electrochemical gradient useful for the energy demanding cellular processes. Most of the oxidases belong to the heme-copper superfamily. They possess a heme-copper center, constituted of a high spin heme and a CuB center, where the reduction of oxygen takes place and probably where the link to proton pumping is located. The superfamily is divided in two classes: the quinol- and the cytochrome c-oxidases. The latter are divided in the aa3 and the cbb3-type cytochrome c oxidases. The main difference between quinol- and the aa3-type cytochrome c-oxidases is the CuA center, which is absent in the quinol oxidases. The cbb3-type cytochrome oxidases have the binuclear center, but lack the CuA center. They also does not have the classical subunits II and III. These differences seem not to affect the oxygen reduction or the proton pumping. Probably the oxidases have evolved from some denitrification enzymes and prior the photosynthetic process. Also is possible that the cbb3-type cytochrome oxidases or others very similar have been the first oxidases to appear.

  20. Identification of the alternative terminal oxidase of higher plant mitochondria

    PubMed Central

    Elthon, Thomas E.; McIntosh, Lee

    1987-01-01

    In addition to cytochrome oxidase, plant mitochondria have a second terminal oxidase called the alternative oxidase. The alternative oxidase is of great interest in that energy is not conserved when electrons flow through it. The potential energy of the system is thus lost as heat, and, in plants with high levels of the alternative oxidase, this results in thermogenesis. We have purified the alternative oxidase from mitochondria of the thermogenic spadix of Sauromatum guttatum and have identified its polypeptide constituents by using polyclonal antibodies. A 166-fold purification was achieved through a combination of cation-exchange (carboxymethyl-Sepharose) and hydrophobic-interaction (phenyl-Sepharose) chromatography. Polyclonal antibodies raised to the CM-Sepharose fractions readily immunoprecipitated alternative oxidase activity and immunoprecipitated four of the proteins that copurify with the activity. These proteins have apparent molecular masses of 37, 36, 35.5, and 35 kDa. Polyclonal antibodies raised individually to the 37-, 36-, and 35.5- plus 35-kDa proteins cross-reacted with all of these proteins, indicating the presence of common antigenic sites. The 37-kDa protein appears to be constitutive in Sauromatum, whereas expression of the 36- and 35-kDa proteins was correlated with presence of alternative pathway activity. The 35.5-kDa protein appears with loss of alternative pathway activity during senescence, indicating that this protein may be a degradation product of the 36-kDa protein. Binding of anti-36-kDa protein antibodies to total mitochondrial protein blots of five plant species indicated that similar proteins were always present when alternative pathway activity was observed. Images PMID:16593898

  1. Molecular and Biochemical Characterization of a Cytokinin Oxidase from Maize1

    PubMed Central

    Bilyeu, Kristin D.; Cole, Jean L.; Laskey, James G.; Riekhof, Wayne R.; Esparza, Thomas J.; Kramer, Michelle D.; Morris, Roy O.

    2001-01-01

    It is generally accepted that cytokinin oxidases, which oxidatively remove cytokinin side chains to produce adenine and the corresponding isopentenyl aldehyde, play a major role in regulating cytokinin levels in planta. Partially purified fractions of cytokinin oxidase from various species have been studied for many years, but have yet to clearly reveal the properties of the enzyme or to define its biological significance. Details of the genomic organization of the recently isolated maize (Zea mays) cytokinin oxidase gene (ckx1) and some of its Arabidopsis homologs are now presented. Expression of an intronless ckx1 in Pichia pastoris allowed production of large amounts of recombinant cytokinin oxidase and facilitated detailed kinetic and cofactor analysis and comparison with the native enzyme. The enzyme is a flavoprotein containing covalently bound flavin adenine dinucleotide, but no detectable heavy metals. Expression of the oxidase in maize tissues is described. PMID:11154345

  2. 21 CFR 866.2420 - Oxidase screening test for gonorrhea.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Oxidase screening test for gonorrhea. 866.2420 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...

  3. 21 CFR 866.2420 - Oxidase screening test for gonorrhea.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Oxidase screening test for gonorrhea. 866.2420 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...

  4. 21 CFR 866.2420 - Oxidase screening test for gonorrhea.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Oxidase screening test for gonorrhea. 866.2420 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...

  5. 21 CFR 866.2420 - Oxidase screening test for gonorrhea.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Oxidase screening test for gonorrhea. 866.2420 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...

  6. 21 CFR 866.2420 - Oxidase screening test for gonorrhea.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Oxidase screening test for gonorrhea. 866.2420 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...

  7. Producing alcohol and other drugs as a policy 'problem': A critical analysis of South Africa's 'National Drug Master Plan' (2013-2017).

    PubMed

    Pienaar, Kiran; Savic, Michael

    2016-04-01

    The strong symbolic value of illicit drug use makes it a contested issue, which attracts mixed public opinion, intense media attention and close political scrutiny. This means that the formulation of plausible, authoritative policies governing illicit drugs must navigate fraught political terrain. In a country like South Africa with its long unique history of institutionalised oppression of the black majority, the issues confronting drug policy are particularly complex and the need for carefully formulated policy responses especially urgent. Yet despite this, the area of drug policy development in South Africa has received little scholarly attention to date. This paper explores the complexities of policymaking in the South African context by drawing on feminist scholar Carol Bacchi's poststructuralist approach to policy analysis, which focuses on how policy helps to produce the problems it purports to solve. Taking as its empirical focus, South Africa's current drug policy, the third National Drug Master Plan (NDMP), 2013-2017, the paper analyses how the policy constitutes the 'problem of alcohol and other drugs' (AODs). We identify three central policy proposals through which specific problematisations emerge: (1) the proposal that drug use is a global issue requiring a coordinated policy response, (2) appeals to evidence-based policy proposals and (3) the proposal that AOD 'use' and 'abuse' be treated interchangeably. We suggest that these proposals reveal a tendency towards inflating the 'problem of AODs' and thus work to justify punitive policy measures. In an effort to explore the implications of particular problematisations for effecting social change, we clarify the ways in which the policy may work to undermine the interests of those it seeks to aid by reinforcing stigma and marginalisation. Copyright © 2015 Elsevier B.V. All rights reserved.

  8. Identification of a Third Mn(II) Oxidase Enzyme in Pseudomonas putida GB-1

    PubMed Central

    Smesrud, Logan; Tebo, Bradley M.

    2016-01-01

    ABSTRACT The oxidation of soluble Mn(II) to insoluble Mn(IV) is a widespread bacterial activity found in a diverse array of microbes. In the Mn(II)-oxidizing bacterium Pseudomonas putida GB-1, two Mn(II) oxidase genes, named mnxG and mcoA, were previously identified; each encodes a multicopper oxidase (MCO)-type enzyme. Expression of these two genes is positively regulated by the response regulator MnxR. Preliminary investigation into putative additional regulatory pathways suggested that the flagellar regulators FleN and FleQ also regulate Mn(II) oxidase activity; however, it also revealed the presence of a third, previously uncharacterized Mn(II) oxidase activity in P. putida GB-1. A strain from which both of the Mn(II) oxidase genes and fleQ were deleted exhibited low levels of Mn(II) oxidase activity. The enzyme responsible was genetically and biochemically identified as an animal heme peroxidase (AHP) with domain and sequence similarity to the previously identified Mn(II) oxidase MopA. In the ΔfleQ strain, P. putida GB-1 MopA is overexpressed and secreted from the cell, where it actively oxidizes Mn. Thus, deletion of fleQ unmasked a third Mn(II) oxidase activity in this strain. These results provide an example of an Mn(II)-oxidizing bacterium utilizing both MCO and AHP enzymes. IMPORTANCE The identity of the Mn(II) oxidase enzyme in Pseudomonas putida GB-1 has been a long-standing question in the field of bacterial Mn(II) oxidation. In the current work, we demonstrate that P. putida GB-1 employs both the multicopper oxidase- and animal heme peroxidase-mediated pathways for the oxidation of Mn(II), rendering this model organism relevant to the study of both types of Mn(II) oxidase enzymes. The presence of three oxidase enzymes in P. putida GB-1 deepens the mystery of why microorganisms oxidize Mn(II) while providing the field with the tools necessary to address this question. The initial identification of MopA as a Mn(II) oxidase in this strain required the

  9. The first mammalian aldehyde oxidase crystal structure: insights into substrate specificity.

    PubMed

    Coelho, Catarina; Mahro, Martin; Trincão, José; Carvalho, Alexandra T P; Ramos, Maria João; Terao, Mineko; Garattini, Enrico; Leimkühler, Silke; Romão, Maria João

    2012-11-23

    Aldehyde oxidases have pharmacological relevance, and AOX3 is the major drug-metabolizing enzyme in rodents. The crystal structure of mouse AOX3 with kinetics and molecular docking studies provides insights into its enzymatic characteristics. Differences in substrate and inhibitor specificities can be rationalized by comparing the AOX3 and xanthine oxidase structures. The first aldehyde oxidase structure represents a major advance for drug design and mechanistic studies. Aldehyde oxidases (AOXs) are homodimeric proteins belonging to the xanthine oxidase family of molybdenum-containing enzymes. Each 150-kDa monomer contains a FAD redox cofactor, two spectroscopically distinct [2Fe-2S] clusters, and a molybdenum cofactor located within the protein active site. AOXs are characterized by broad range substrate specificity, oxidizing different aldehydes and aromatic N-heterocycles. Despite increasing recognition of its role in the metabolism of drugs and xenobiotics, the physiological function of the protein is still largely unknown. We have crystallized and solved the crystal structure of mouse liver aldehyde oxidase 3 to 2.9 Å. This is the first mammalian AOX whose structure has been solved. The structure provides important insights into the protein active center and further evidence on the catalytic differences characterizing AOX and xanthine oxidoreductase. The mouse liver aldehyde oxidase 3 three-dimensional structure combined with kinetic, mutagenesis data, molecular docking, and molecular dynamics studies make a decisive contribution to understand the molecular basis of its rather broad substrate specificity.

  10. Group motivational interviewing for homeless young adults: Associations of change talk with substance use and sexual risk behavior.

    PubMed

    D'Amico, Elizabeth J; Houck, Jon M; Tucker, Joan S; Ewing, Brett A; Pedersen, Eric R

    2017-09-01

    Homeless young adults exhibit high rates of alcohol and other drug (AOD) use and sexual risk behaviors. This study is a secondary analysis of data collected in a randomized clinical trial of AWARE, a new 4 session group motivational interviewing intervention. AWARE mainly focused on alcohol use and sexual risk behavior given focus group feedback. We used sequential coding to analyze how the group process affected both AOD use and sexual risk behavior at 3-month follow up among homeless young adults by examining facilitator behavior and participant change talk (CT) and sustain talk (ST). We analyzed 57 group session digital recordings of 100 youth (69% male, 74% heterosexual, 28% non-Hispanic white, 23% African American, 26% Hispanic, 23% multiracial/other; mean age 21.75). Outcomes included importance and readiness to change AOD use and risky sexual behavior, AOD use and consequences, number of partners and unprotected sex, and condom self-efficacy. Sequential analysis indicated that facilitator open-ended questions and reflections of CT increased Group CT. Group CT was associated with a lower likelihood of being a heavy drinker 3 months later; Group ST was associated with decreased readiness and confidence to change alcohol use. There were no associations with CT or ST for drug use or risky sexual behavior. Facilitator speech and peer responses were related to CT and ST during the group sessions with this high risk population, which were then associated with individual changes for alcohol use. Further research is needed to explore associations with drug use and sexual risk behavior. (PsycINFO Database Record (c) 2017 APA, all rights reserved).

  11. [Oxygen and the superoxide anion. Modulation of NADPH oxidase?].

    PubMed

    Delbosc, S; Cristol, J P; Descomps, B; Chénard, J; Sirois, P

    2001-01-01

    Oxidative stress which results from an imbalance between oxidant production and antioxidant defense mechanisms can promote modifications of lipids, proteins and nucleic acids. This review focuses on the different pathways leading to Reactive Oxygen Species (ROS) production in particular on NADPH oxidase activation. This enzyme is localized in numerous cells including phagocytes and vascular cells and composed of membrane and cytosolic sub-units. The activation of the NADPH oxidase is largely involved in inflammation associated diseases such as asthma, Systemic Inflammatory Response Syndrome and aging associated diseases such as atherosclerosis and neurodeneratives diseases. The modulation of NADPH oxidase could be a way to limit or prevent the development of these diseases.

  12. Aiding and abetting roles of NOX oxidases in cellular transformation

    PubMed Central

    Block, Karen; Gorin, Yves

    2013-01-01

    NADPH oxidases of the NADPH oxidase (NOX) family are dedicated reactive oxygen species-generating enzymes that broadly and specifically regulate redox-sensitive signalling pathways that are involved in cancer development and progression. They act at specific cellular membranes and microdomains through the activation of oncogenes and the inactivation of tumour suppressor proteins. In this Review, we discuss primary targets and redox-linked signalling systems that are influenced by NOX-derived ROS, and the biological role of NOX oxidases in the aetiology of cancer. PMID:22918415

  13. Methods for transfer a saliva based alcohol content test to a dermal patch

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

    Silks, III, Louis A.

    Detection and quantitation of ethanol which is highly sensitive, specific, and efficient has been a commercial target for sometime. Clearly analytical methods are useful such as gas and liquid chromatography, mass spectrometry, and NMR spectroscopy. However, those methods are best used in the laboratory and a less useful for detection and quantitation of ethanol in the field. Enzymes have been employed for the detection and quantitation of EtOH. Enzymes are proteins that perform a particular task in a bio-catalytic way. Most of the chemistry that these enzymes do are frequently exquisitely specific in that only one alcohol reacts and onlymore » one product is produced. One enzyme molecule can catalyze the reaction of numerous substrate molecules which in itself is an amplification of the recognition signal. Alcohol dehydrogenase (ADH) and alcohol oxidase (AO) are two possible enzymatic targets for EtOH sensor development.1 The ADH oxidizes the alcohol using a co-factor nicotinamide adenine dinucleotide. This co-factor needs to be within close proximity of the ADH. AO also oxidizes the ethanol using molecular oxygen giving rise to the production of the aldehyde and hydrogen peroxide.« less

  14. NADPH Oxidase Activation Contributes to Heavy Ion Irradiation–Induced Cell Death

    PubMed Central

    Wang, Yupei; Liu, Qing; Zhao, Weiping; Zhou, Xin; Miao, Guoying; Sun, Chao

    2017-01-01

    Increased oxidative stress plays an important role in heavy ion radiation–induced cell death. The mechanism involved in the generation of elevated reactive oxygen species (ROS) is not fully illustrated. Here we show that NADPH oxidase activation is closely related to heavy ion radiation–induced cell death via excessive ROS generation. Cell death and cellular ROS can be greatly reduced in irradiated cancer cells with the preincubation of diphenyleneiodium, an inhibitor of NADPH oxidase. Most of the NADPH oxidase (NOX) family proteins (NOX1, NOX2, NOX3, NOX4, and NOX5) showed increased expression after heavy ion irradiation. Meanwhile, the cytoplasmic subunit p47phox was translocated to the cell membrane and localized with NOX2 to form reactive NADPH oxidase. Our data suggest for the first time that ROS generation, as mediated by NADPH oxidase activation, could be an important contributor to heavy ion irradiation–induced cell death. PMID:28473742

  15. Immobilization of xanthine oxidase on a polyaniline silicone support.

    PubMed

    Nadruz, W; Marques, E T; Azevedo, W M; Lima-Filho, J L; Carvalho, L B

    1996-03-01

    A polyaniline silicone support to immobilize xanthine oxidase is proposed as a reactor coil to monitor the action of xanthine oxidase on hypoxanthine, xanthine and 6-mercaptopurine. A purified xanthine oxidase immobilized on this support lost 80% of the initial activity after 12 min of use. Co-immobilization of superoxide dismutase and catalase increased the stability of immobilized xanthine oxidase so that the derivative maintained 79% of its initial activity after 4.6 h of continuous use in which 1.5 mumol purine bases were converted by the immobilized enzyme system. There is no evidence of either polyaniline or protein leaching from the coil during 3 h of continuous use. When solutions (10 ml) of hypoxanthine, xanthine and 6-mercaptopurine were circulated individually through the xanthine oxidase-superoxide dismutase-catalase-polyaniline coil (1 mm internal diameter and 3 m in length, 3 ml internal volume) activities of 8.12, 11.17 and 1.09 nmol min-1 coil-1, respectively, were obtained. The advantages of the reactor configuration and the redox properties of the polymer, particularly with respect to immobilized oxidoreductases, make this methodology attractive for similar enzyme systems. This immobilized enzyme system using polyaniline-silicone as support converted 6-mercaptopurine to 6-thiouric acid with equal efficiency as resins based on polyacrylamide and polyamide 11.

  16. Cytotoxicity of polyamines to Amoeba proteus: role of polyamine oxidase.

    PubMed

    Schenkel, E; Dubois, J G; Helson-Cambier, M; Hanocq, M

    1996-02-01

    It has been shown that oxidation of polyamines by polyamine oxidases can produce toxic compounds (H2O2, aldehydes, ammonia) and that the polyamine oxidase-polyamine system is implicated, in vitro, in the death of several parasites. Using Amoeba proteus as an in vitro model, we studied the cytotoxicity to these cells of spermine, spermidine, their acetyl derivatives, and their hypothetical precursors. Spermine and N1-acetylspermine were more toxic than emetine, an amoebicidal reference drug. Spermine presented a short-term toxicity, but a 48-h contact time was necessary for the high toxicity of spermidine. The uptake by Amoeba cells of the different polyamines tested was demonstrated. On the other hand, a high polyamine oxidase activity was identified in Amoeba proteus crude extract. Spermine (theoretical 100%) and N1-acetylspermine (64%) were the best substrates at pH 9.5, while spermidine, its acetyl derivatives, and putrescine were very poorly oxidized by this enzyme (3-20%). Spermine oxidase activity was inhibited by phenylhydrazine (nil) and isoniazid (approximately 50%). Mepacrine did not inhibit the enzyme activity at pH 8. Neither monoamine nor diamine oxidase activity (approximately 10%) was found. It must be emphasized that spermine, the best enzyme substrate, is the most toxic polyamine. This finding suggests that knowledge of polyamine oxidase specificity can be used to modulate the cytotoxicity of polyamine derivatives. Amoeba proteus was revealed as a simple model for investigation of the connection between cytotoxicity and enzyme activity.

  17. [The regulation of peroxisomal matrix enzymes (alcohol oxidase and catalase) formation by the product of the gene Mth1 in methylotrophic yeast Pichia methanolica].

    PubMed

    Leonovich, O A; Kurales, Iu A; Dutova, T A; Isakova, E P; Deriabina, Iu I; Rabinovich, Ia M

    2009-01-01

    Two independent mutant strains of methylotrophic yeast Pichia methanolica (mth1 arg1 and mth2 arg4) from the initial line 616 (ade1 ade5) were investigated. The mutant strains possessed defects in genes MTH1 and MTH2 which resulted in the inability to assimilate methanol as a sole carbon source and the increased activity of alcohol oxidase (AO). The function of the AUG2 gene encoding one of the subunits of AO and CTA1, a probable homolog of peroxisomal catalase of Saccharomyces cereviseae, was investigated by analyses of the molecular forms of isoenzymes. It was shown that optimal conditions for the expression of the AUG2 gene on a medium supplemented with 3% of methanol leads to an increasing synthesis of peroxisomal catalase. The mutant mth1 possessed a dominant formation of AO isoform with electrophoretic mobility which is typical for isogenic form 9, the product of the AUG2 gene, and a decreased level of peroxisomal catalase. The restoration of growth of four spontaneous revertants of the mutant mth1 (Rmth1) on the methanol containing medium was accompanied by an increase in activity of AO isogenic form 9 and peroxisomal catalase. The obtained results confirmed the functional continuity of the structural gene AUG2 in mutant mth1. The correlation of activity of peroxisomal catalase and AO isogenic form 1 in different conditions evidenced the existence of common regulatory elements for genes AUG2 and CTA1 in methilotrophic yeast Pichia methanolica.

  18. Simple, high-yield purification of xanthine oxidase from bovine milk.

    PubMed

    Ozer, N; Müftüoglu, M; Ataman, D; Ercan, A; Ogüs, I H

    1999-05-13

    Xanthine oxidase, a commercially important enzyme with a wide area of application, was extracted from fresh milk, without added preservatives, using toluene and heat. The short purification procedure, with high yield, consisted of extraction, ammonium sulfate fractionation, and DEAE-Sepharose (fast flow) column chromatography. Xanthine oxidase was eluted as a single activity peak from the column using a buffer gradient. The purification fold, specific activity and yield for the purified xanthine oxidase were 328, 10.161 U/mg and 69%, respectively. The enzyme was concentrated by ultrafiltration, although 31% of the activity was lost during concentration, no change in specific activity was observed. Activity and protein gave coincident staining bands on native polyacrylamide gels. The intensity and the number of bands were dependent on the oxidative state(s) of the enzyme; reduction by 2-mercaptoethanol decreased the intensity of the slow-moving bands and increased the intensity of the fastest-moving band. Following sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), two major bands (molecular masses of 152 and 131 kDa) were observed, accounting for > or = 95% of xanthine oxidase. Native- and SDS-PAGE showed that the purified xanthine oxidase becomes a heterodimer due to endogenous proteases.

  19. Urate Oxidase Purification by Salting-in Crystallization: Towards an Alternative to Chromatography

    PubMed Central

    Giffard, Marion; Ferté, Natalie; Ragot, François; El Hajji, Mohamed; Castro, Bertrand; Bonneté, Françoise

    2011-01-01

    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 changes to the

  20. Construction of Mutant Glucose Oxidases with Increased Dye-Mediated Dehydrogenase Activity

    PubMed Central

    Horaguchi, Yohei; Saito, Shoko; Kojima, Katsuhiro; Tsugawa, Wakako; Ferri, Stefano; Sode, Koji

    2012-01-01

    Mutagenesis studies on glucose oxidases (GOxs) were conducted to construct GOxs with reduced oxidase activity and increased dehydrogenase activity. We focused on two representative GOxs, of which crystal structures have already been reported—Penicillium amagasakiense GOx (PDB ID; 1gpe) and Aspergillus niger GOx (PDB ID; 1cf3). We constructed oxygen-interacting structural models for GOxs, and predicted the residues responsible for oxidative half reaction with oxygen on the basis of the crystal structure of cholesterol oxidase as well as on the fact that both enzymes are members of the glucose/methanol/choline (GMC) oxidoreductase family. Rational amino acid substitution resulted in the construction of an engineered GOx with drastically decreased oxidase activity and increased dehydrogenase activity, which was higher than that of the wild-type enzyme. As a result, the dehydrogenase/oxidase ratio of the engineered enzyme was more than 11-fold greater than that of the wild-type enzyme. These results indicate that alteration of the dehydrogenase/oxidase activity ratio of GOxs is possible by introducing a mutation into the putative functional residues responsible for oxidative half reaction with oxygen of these enzymes, resulting in a further increased dehydrogenase activity. This is the first study reporting the alteration of GOx electron acceptor preference from oxygen to an artificial electron acceptor. PMID:23203056

  1. Structure of caa(3) cytochrome c oxidase--a nature-made enzyme-substrate complex.

    PubMed

    Noor, Mohamed Radzi; Soulimane, Tewfik

    2013-05-01

    Aerobic respiration, the energetically most favorable metabolic reaction, depends on the action of terminal oxidases that include cytochrome c oxidases. The latter forms a part of the heme-copper oxidase superfamily and consists of three different families (A, B, and C types). The crystal structures of all families have now been determined, allowing a detailed structural comparison from evolutionary and functional perspectives. The A2-type oxidase, exemplified by the Thermus thermophilus caa(3) oxidase, contains the substrate cytochrome c covalently bound to the enzyme complex. In this article, we highlight the various features of caa(3) enzyme and provide a discussion of their importance, including the variations in the proton and electron transfer pathways.

  2. The Importance of NADPH Oxidases and Redox Signaling in Angiogenesis

    PubMed Central

    Prieto-Bermejo, Rodrigo; Hernández-Hernández, Angel

    2017-01-01

    Eukaryotic cells have to cope with the constant generation of reactive oxygen species (ROS). Although the excessive production of ROS might be deleterious for cell biology, there is a plethora of evidence showing that moderate levels of ROS are important for the control of cell signaling and gene expression. The family of the nicotinamide adenine dinucleotide phosphate oxidases (NADPH oxidases or Nox) has evolved to produce ROS in response to different signals; therefore, they fulfil a central role in the control of redox signaling. The role of NADPH oxidases in vascular physiology has been a field of intense study over the last two decades. In this review we will briefly analyze how ROS can regulate signaling and gene expression. We will address the implication of NADPH oxidases and redox signaling in angiogenesis, and finally, the therapeutic possibilities derived from this knowledge will be discussed. PMID:28505091

  3. Degradation of oxalate in rats implanted with immobilized oxalate oxidase.

    PubMed

    Raghavan, K G; Tarachand, U

    1986-01-20

    Accumulation of oxalate leads to hyperoxaluria and calcium oxalate nephrolithiasis in man. Since oxalate is a metabolic end product in mammals, the feasibility of its enzymic degradation has been tested in vivo in rats by administering exogenous oxalate oxidase. Oxalate oxidase, isolated from banana fruit peels, in its native form was found to be non-active at the physiological pH of the recipient animal. However, its functional viability in the recipient animal was ensured by its prior binding with ethylenemaleic anhydride, thus shifting its pH activity curve towards the alkaline range. Rats implanted with dialysis membrane capsules containing such immobilized oxalate oxidase in their peritoneal cavities effectively metabolized intraperitoneally injected [14C]oxalate as well as its precursor [14C]glyoxalate. The implantation of capsules containing coentrapped multienzyme preparations of oxalate oxidase, catalase and peroxidase led to a further degradation of administered [14C]oxalate in rats.

  4. Urate oxidase is imported into peroxisomes recognizing the C-terminal SKL motif of proteins.

    PubMed

    Miura, S; Oda, T; Funai, T; Ito, M; Okada, Y; Ichiyama, A

    1994-07-01

    Rat liver urate oxidase synthesized from cDNA through coupled transcription and translation was incubated at 26 degrees C for 60 min with purified peroxisomes from rat liver. Urate oxidase was efficiently imported into the peroxisomes, as determined by resistance to externally added proteinase K. The amount of imported urate oxidase increased with time and the import was temperature dependent. A synthetic peptide composed of the C-terminal 10 amino acid residues of acyl-CoA oxidase (the C-terminal tripeptide is Ser-Lys-Leu) inhibited the import of urate oxidase, whereas other peptides, in which the C-terminal Ser-Lys-Leu (SKL) sequence was deleted or mutated, were not effective. Two mutant urate oxidase proteins in which the C-terminal Ser-Arg-Leu (SRL) sequence was deleted or mutated to Ser-Glu-Leu (SEL) were not imported into peroxisomes. With substitution of a lysine residue for arginine in the SRL tripeptide at the C-terminus the import activity was retained. These results show that urate oxidase is important into peroxisomes via a common pathway with acyl-CoA oxidase, and that the C-terminal SRL sequence functions as a peroxisomal-targeting signal.

  5. Multilayered Polyelectrolyte Microcapsules: Interaction with the Enzyme Cytochrome C Oxidase

    PubMed Central

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

    2014-01-01

    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. PMID:25372607

  6. Identification in Marinomonas mediterranea of a novel quinoprotein with glycine oxidase activity.

    PubMed

    Campillo-Brocal, Jonatan Cristian; Lucas-Elio, Patricia; Sanchez-Amat, Antonio

    2013-08-01

    A novel enzyme with lysine-epsilon oxidase activity was previously described in the marine bacterium Marinomonas mediterranea. This enzyme differs from other l-amino acid oxidases in not being a flavoprotein but containing a quinone cofactor. It is encoded by an operon with two genes lodA and lodB. The first one codes for the oxidase, while the second one encodes a protein required for the expression of the former. Genome sequencing of M. mediterranea has revealed that it contains two additional operons encoding proteins with sequence similarity to LodA. In this study, it is shown that the product of one of such genes, Marme_1655, encodes a protein with glycine oxidase activity. This activity shows important differences in terms of substrate range and sensitivity to inhibitors to other glycine oxidases previously described which are flavoproteins synthesized by Bacillus. The results presented in this study indicate that the products of the genes with different degrees of similarity to lodA detected in bacterial genomes could constitute a reservoir of different oxidases. © 2013 The Authors. Microbiology Open published by John Wiley & Sons Ltd.

  7. Functional expression of amine oxidase from Aspergillus niger (AO-I) in Saccharomyces cerevisiae.

    PubMed

    Kolaríková, Katerina; Galuszka, Petr; Sedlárová, Iva; Sebela, Marek; Frébort, Ivo

    2009-01-01

    The aim of this work was to prepare recombinant amine oxidase from Aspergillus niger after overexpressing in yeast. The yeast expression vector pDR197 that includes a constitutive PMA1 promoter was used for the expression in Saccharomyces cerevisiae. Recombinant amine oxidase was extracted from the growth medium of the yeast, purified to homogeneity and identified by activity assay and MALDI-TOF peptide mass fingerprinting. Similarity search in the newly published A. niger genome identified six genes coding for copper amine oxidase, two of them corresponding to the previously described enzymes AO-I a methylamine oxidase and three other genes coding for FAD amine oxidases. Thus, A. niger possesses an enormous metabolic gear to grow on amine compounds and thus support its saprophytic lifestyle.

  8. Cyanide-insensitive quinol oxidase (CIO) from Gluconobacter oxydans is a unique terminal oxidase subfamily of cytochrome bd.

    PubMed

    Miura, Hiroshi; Mogi, Tatsushi; Ano, Yoshitaka; Migita, Catharina T; Matsutani, Minenosuke; Yakushi, Toshiharu; Kita, Kiyoshi; Matsushita, Kazunobu

    2013-06-01

    Cyanide-insensitive terminal quinol oxidase (CIO) is a subfamily of cytochrome bd present in bacterial respiratory chain. We purified CIO from the Gluconobacter oxydans membranes and characterized its properties. The air-oxidized CIO showed some or weak peaks of reduced haemes b and of oxygenated and ferric haeme d, differing from cytochrome bd. CO- and NO-binding difference spectra suggested that haeme d serves as the ligand-binding site of CIO. Notably, the purified CIO showed an extraordinary high ubiquinol-1 oxidase activity with the pH optimum of pH 5-6. The apparent Vmax value of CIO was 17-fold higher than that of G. oxydans cytochrome bo3. In addition, compared with Escherichia coli cytochrome bd, the quinol oxidase activity of CIO was much more resistant to cyanide, but sensitive to azide. The Km value for O2 of CIO was 7- to 10-fold larger than that of G. oxydans cytochrome bo3 or E. coli cytochrome bd. Our results suggest that CIO has unique features attributable to the structure and properties of the O2-binding site, and thus forms a new sub-group distinct from cytochrome bd. Furthermore, CIO of acetic acid bacteria may play some specific role for rapid oxidation of substrates under acidic growth conditions.

  9. Globular adiponectin inhibits ethanol-induced reactive oxygen species production through modulation of NADPH oxidase in macrophages: involvement of liver kinase B1/AMP-activated protein kinase pathway.

    PubMed

    Kim, Mi Jin; Nagy, Laura E; Park, Pil-Hoon

    2014-09-01

    Adiponectin, an adipokine predominantly secreted from adipocytes, has been shown to play protective roles against chronic alcohol consumption. Although excessive reactive oxygen species (ROS) production in macrophages is considered one of the critical events for ethanol-induced damage in various target tissues, the effect of adiponectin on ethanol-induced ROS production is not clearly understood. In the present study, we investigated the effect of globular adiponectin (gAcrp) on ethanol-induced ROS production and the potential mechanisms underlying these effects of gAcrp in macrophages. Here we demonstrated that gAcrp prevented ethanol-induced ROS production in both RAW 264.7 macrophages and primary murine peritoneal macrophages. Globular adiponectin also inhibited ethanol-induced activation of NADPH oxidase. In addition, gAcrp suppressed ethanol-induced increase in the expression of NADPH oxidase subunits, including Nox2 and p22(phox), via modulation of nuclear factor-κB pathway. Furthermore, pretreatment with compound C, a selective inhibitor of AMPK, or knockdown of AMPK by small interfering RNA restored suppression of ethanol-induced ROS production and Nox2 expression by gAcrp. Finally, we found that gAcrp treatment induced phosphorylation of liver kinase B1 (LKB1), an upstream signaling molecule mediating AMPK activation. Knockdown of LKB1 restored gAcrp-suppressed Nox2 expression, suggesting that LKB1/AMPK pathway plays a critical role in the suppression of ethanol-induced ROS production and activation of NADPH oxidase by gAcrp. Taken together, these results demonstrate that globular adiponectin prevents ethanol-induced ROS production, at least in part, via modulation of NADPH oxidase in macrophages. Further, LKB1/AMPK axis plays an important role in the suppression of ethanol-induced NADPH oxidase activation by gAcrp in macrophages. Copyright © 2014 by The American Society for Pharmacology and Experimental Therapeutics.

  10. Evidence for proton tunneling and a transient covalent flavin-substrate adduct in choline oxidase S101A.

    PubMed

    Uluisik, Rizvan; Romero, Elvira; Gadda, Giovanni

    2017-11-01

    The effect of temperature on the reaction of alcohol oxidation catalyzed by choline oxidase was investigated with the S101A variant of choline oxidase. Anaerobic enzyme reduction in a stopped-flow spectrophotometer was biphasic using either choline or 1,2-[ 2 H 4 ]-choline as a substrate. The limiting rate constants k lim1 and k lim2 at saturating substrate were well separated (k lim1 /k lim2 >9), and were >15-fold slower than for wild-type choline oxidase. Solvent deuterium kinetic isotope effects (KIEs) ~4 established that k lim1 probes the proton transfer from the substrate hydroxyl to a catalytic base. Primary substrate deuterium KIEs ≥7 demonstrated that k lim2 reports on hydride transfer from the choline alkoxide to the flavin. Between 15°C and 39°C the k lim1 and k lim2 values increased with increasing temperature, allowing for the analyses of H + and H - transfers using Eyring and Arrhenius formalisms. Temperature-independent KIE on the k lim1 value ( H2O k lim1 / D2O k lim1 ) suggests that proton transfer occurs within a highly reorganized tunneling-ready-state with a narrow distribution of donor-acceptor distances. Eyring analysis of the k lim2 value gave lines with the slope (choline) >slope (D-choline) , suggesting kinetic complexity. Spectral evidence for the transient occurrence of a covalent flavin-substrate adduct during the first phase of the anaerobic reaction of S101A CHO with choline is presented, supporting the notion that an important role of amino acid residues in the active site of flavin-dependent enzymes is to eliminate alternative reactions of the versatile enzyme-bound flavin for the reaction that needs to be catalyzed. Copyright © 2017 Elsevier B.V. All rights reserved.

  11. Determination of Monoamine Oxidase A and B Activity in Long-Term Treated Patients With Parkinson Disease.

    PubMed

    Müller, Thomas; Riederer, Peter; Grünblatt, Edna

    Biogenic amines and monoamine oxidase inhibitors influence peripheral monoamine oxidase enzyme activity in chronic levodopa/dopa decarboxylase inhibitor-treated patients with Parkinson disease. Rasagiline is an irreversible inhibitor of monoamine oxidase B. Safinamide blocks this isoenzyme in a reversible fashion. The aim of this study was to determine monoamine oxidase A (plasma) and B (platelets) enzyme activity in long-term levodopa-treated patients without and with additional oral intake of 50- or 100-mg safinamide or 1-mg rasagiline or first-time intake of rasagiline. Monoamine oxidase A enzyme activity did not differ between all groups. Patients on rasagiline or safinamide showed lower monoamine oxidase-B enzyme activity compared with patients without monoamine oxidase B inhibitor intake. No impact of the number of previous oral levodopa intakes was found. Rasagiline and safinamide did not essentially differ in terms of inhibition of monoamine oxidase B despite their different pharmacology regarding reversibility of monoamine oxidase B inhibition. In view of the observed, considerable heterogeneity of enzyme activities, we suggest to determine activities of monoamine oxidase A and B to reduce the risk for tyramine-induced hypertension and the serotonergic syndrome during chronic therapy with rasagiline or safinamide.

  12. Platinum Nanoparticles: Efficient and Stable Catechol Oxidase Mimetics.

    PubMed

    Liu, Yi; Wu, Haohao; Chong, Yu; Wamer, Wayne G; Xia, Qingsu; Cai, Lining; Nie, Zhihong; Fu, Peter P; Yin, Jun-Jie

    2015-09-09

    Although enzyme-like nanomaterials have been extensively investigated over the past decade, most research has focused on the peroxidase-like, catalase-like, or SOD-like activity of these nanomaterials. Identifying nanomaterials having oxidase-like activities has received less attention. In this study, we demonstrate that platinum nanoparticles (Pt NPs) exhibit catechol oxidase-like activity, oxidizing polyphenols into the corresponding o-quinones. Four unique approaches are employed to demonstrate the catechol oxidase-like activity exerted by Pt NPs. First, UV-vis spectroscopy is used to monitor the oxidation of polyphenols catalyzed by Pt NPs. Second, the oxidized products of polyphenols are identified by ultrahigh-performance liquid chromatography (UHPLC) separation followed by high-resolution mass spectrometry (HRMS) identification. Third, electron spin resonance (ESR) oximetry techniques are used to confirm the O2 consumption during the oxidation reaction. Fourth, the intermediate products of semiquinone radicals formed during the oxidation of polyphenols are determined by ESR using spin stabilization. These results indicate Pt NPs possess catechol oxidase-like activity. Because polyphenols and related bioactive substances have been explored as potent antioxidants that could be useful for the prevention of cancer and cardiovascular diseases, and Pt NPs have been widely used in the chemical industry and medical science, it is essential to understand the potential effects of Pt NPs for altering or influencing the antioxidant activity of polyphenols.

  13. PROLINE OXIDASES IN HANSENULA SUBPELLICULOSA

    PubMed Central

    Ling, Chung-Mei; Hedrick, L. R.

    1964-01-01

    Ling, Chung-Mei (Illinois Institute of Technology, Chicago), and L. R. Hedrick. Proline oxidases in Hansenula subpelliculosa. J. Bacteriol. 87:1462–1470. 1964—Cells of Hansenula subpelliculosa can use l-proline as a carbon and a nitrogen source after a 6- to 8-hr induction period. However, they cannot use l-glutamate as both nitrogen and carbon sources unless the induction period is of several days' duration. Two l-proline oxidases were demonstrated in the mitochondrial preparation of this yeast. One forms the product Δ′-pyrroline-2-carboxylic acid (P2C), which is in equilibrium with α-keto-δ-amino-valeric acid; the other forms the product Δ′-pyrroline-5-carboxylic acid (P5C), which is in equilibrium with glutamic-γ-semialdehyde. The first-mentioned enzyme is induced when l-proline is the carbon source; the second appears to be constitutive, and is probably associated with the use of l-proline as a nitrogen source. The P2C-forming enzyme is specific for the l isomer of proline, and is inactive against l-hydroxyproline. The enzyme activity is at its peak when the mitochondria are prepared from logarithmically grown cells, and is rapidly reduced after cells reach the stationary phase of growth. Kinetic studies with varying concentrations of substrate indicate a Michaelis-Menten constant of 2.45 × 10−2m. Paper chromatographic studies, chemical tests with H2O2, sensitivity to freezing, and spectral measurements indicate that proline oxidase from H. subpelliculosa mitochondria forms a product from l-proline which is like, if not identical to, P2C formed by the action of sheep kidney d-proline oxidase upon dl-proline. The soluble portion of the cell extract contains NAD+ enzymes which use either P2C (α-keto-δ-amino-valeric acid) or P5C (glutamic-γ-semialdehyde) as substrates. No glutamic dehydrogenase activity could be detected when l-glutamic acid and the nicotinamide adenine dinucleotide (NAD+) cofactor were added to the supernatant solution with the

  14. The increasing role of monoamine oxidase type B inhibitors in Parkinson's disease therapy.

    PubMed

    Elmer, Lawrence W; Bertoni, John M

    2008-11-01

    The role of monoamine oxidase type B inhibitors in the treatment of Parkinson's disease has expanded with the new monoamine oxidase B inhibitor rasagiline and a new formulation, selegiline oral disintegrating tablets. As primary therapy in early disease monoamine oxidase B inhibitors reduce motor disability and delay the need for levodopa. In more advanced disease requiring levodopa, adjunctive monoamine oxidase B inhibitors reduce 'off' time and may improve gait and freezing. Rasagiline and selegiline oral disintegrating tablets may reduce the safety risks associated with the amfetamine and methamfetamine metabolites of conventional oral selegiline while retaining or improving therapeutic efficacy. Articles were identified by searches of PubMed and searches on the Internet and reviewed. All articles and other referenced materials were retrieved using the keywords 'Parkinson's disease', 'treatment' and 'monoamine oxidase B inhibitor' and were published between 1960 and 2007, with older references selected for historical significance. Only papers published in English were reviewed. Accumulating data support the use of monoamine oxidase B inhibitors as monotherapy for early and mild Parkinson's disease and as adjunctive therapy for more advanced Parkinson's disease with levodopa-associated motor fluctuations. The recently released monoamine oxidase B inhibitor rasagiline and a new formulation, selegiline oral disintegrating tablets, have potential advantages over conventional oral selegiline.

  15. Photoaffinity labeling of protoporphyrinogen oxidase, the molecular target of diphenylether-type herbicides.

    PubMed

    Camadro, J M; Matringe, M; Thome, F; Brouillet, N; Mornet, R; Labbe, P

    1995-05-01

    Diphenylether-type herbicides are extremely potent inhibitors of protoporphyrinogen oxidase, a membrane-bound enzyme involved in the heme and chlorophyll biosynthesis pathways. Tritiated acifluorfen and a diazoketone derivative of tritiated acifluorfen were specifically bound to a single class of high-affinity binding sites on yeast mitochondrial membranes with apparent dissociation constants of 7 nM and 12.5 nM, respectively. The maximum density of specific binding sites, determined by Scatchard analysis, was 3 pmol.mg-1 protein. Protoporphyrinogen oxidase specific activity was estimated to be 2500 nmol protoporphyrinogen oxidized h-1.mol-1 enzyme. The diazoketone derivative of tritiated acifluorfen was used to specifically photolabel yeast protoporphyrinogen oxidase. The specifically labeled polypeptide in wild-type mitochondrial membranes had an apparent molecular mass of 55 kDa, identical to the molecular mass of the purified enzyme. This photolabeled polypeptide was not detected in a protoporphyrinogen-oxidase-deficient yeast strain, but the membranes contained an equivalent amount of inactive immunoreactive protoporphyrinogen oxidase protein.

  16. Characterization of a Highly Thermostable and Organic Solvent-Tolerant Copper-Containing Polyphenol Oxidase with Dye-Decolorizing Ability from Kurthia huakuii LAM0618T

    PubMed Central

    Guo, Xiang; Zhou, Shan; Wang, Yanwei; Song, Jinlong; Wang, Huimin; Kong, Delong; Zhu, Jie; Dong, Weiwei; He, Mingxiong; Hu, Guoquan; Ruan, Zhiyong

    2016-01-01

    Laccases are green biocatalysts that possess attractive advantages for the treatment of resistant environmental pollutants and dye effluents. A putative laccase-like gene, laclK, encoding a protein of 29.3 kDa and belonging to the Cu-oxidase_4 superfamily, was cloned and overexpressed in Escherichia coli. The purified recombinant protein LaclK (LaclK) was able to oxidize typical laccase substrates such as 2,6-dimethoxyphenol and l-dopamine. The characteristic adsorption maximums of typical laccases at 330 nm and 610 nm were not detected for LaclK. Cu2+ was essential for substrate oxidation, but the ratio of copper atoms/molecule of LaclK was determined to only be 1:1. Notably, the optimal temperature of LaclK was 85°C with 2,6-dimethoxyphenol as substrates, and the half-life approximately 3 days at 80°C. Furthermore, 10% (v/v) organic solvents (methanol, ethanol, isopropyl alcohol, butyl alcohol, Triton x-100 or dimethyl sulfoxide) could promote enzymatic activity. LaclK exhibited wide-spectrum decolorization ability towards triphenylmethane dyes, azo dyes and aromatic dyes, decolorizing 92% and 94% of Victoria Blue B (25 μM) and Ethyl Violet (25 μM), respectively, at a concentration of 60 U/L after 1 h of incubation at 60°C. Overall, we characterized a novel thermostable and organic solvent-tolerant copper-containing polyphenol oxidase possessing dye-decolorizing ability. These unusual properties make LaclK an alternative for industrial applications, particularly processes that require high-temperature conditions. PMID:27741324

  17. Amine oxidase from lentil seedlings: energetic domains and effect of temperature on activity.

    PubMed

    Moosavi-Nejad, S Z; Rezaei-Tavirani, M; Padiglia, A; Floris, G; Moosavi-Movahedi, A A

    2001-07-01

    Copper/TPQ amine oxidases from mammalian and plant sources have shown many differences in substrate specificity and molecular properties. In this work the activity of lentil seedling amine oxidase was followed at various temperatures in 100 mM potassium phosphate buffer, pH 7, using benzylamine as substrate. The discontinuous Arrhenius plot of lentil amine oxidase showed two distinct phases with a jump between them. Thermal denaturation of the enzyme, using differential scanning calorimetry under the same experimental conditions, showed a transition at the same temperature ranges in the absence of substrate, indicating the occurrence of conformational changes, with an enthalpy change of about 175.9 kJ/mole. The temperature-induced changes of the activity of lentil amine oxidase are compared with those of bovine serum amine oxidase (taken from the literature).

  18. Purification of the Alpha Glycerophosphate Oxidase from African Trypanosomes

    DTIC Science & Technology

    1987-02-02

    oxidase (GPO). This enzyme has not been purified or characterize in detail. Inhibition of this enzyme coupled with inhibition of the anaerobic...more manageable afterwards and remained in the procedure although it only slightly increased the yield. The stability of the solubilized enzyme was...whether the detergent was added during the assay or in the solubilization procedure. However, the successful assay for the enzyme was ubiquinol oxidase

  19. Averting the perfect storm: addressing youth substance use risk from social media use.

    PubMed

    Salimian, Parissa K; Chunara, Rumi; Weitzman, Elissa R

    2014-10-01

    Adolescents are developmentally sensitive to pathways that influence alcohol and other drug (AOD) use. In the absence of guidance, their routine engagement with social media may add a further layer of risk. There are several potential mechanisms for social media use to influence AOD risk, including exposure to peer portrayals of AOD use, socially amplified advertising, misinformation, and predatory marketing against a backdrop of lax regulatory systems and privacy controls. Here the authors summarize the influences of the social media world and suggest how pediatricians in everyday practice can alert youth and their parents to these risks to foster conversation, awareness, and harm reduction. Copyright 2014, SLACK Incorporated.

  20. NADPH oxidase mediates depressive behavior induced by chronic stress in mice.

    PubMed

    Seo, Ji-Seon; Park, Jin-Young; Choi, Juli; Kim, Tae-Kyung; Shin, Joo-Hyun; Lee, Ja-Kyeong; Han, Pyung-Lim

    2012-07-11

    Stress is a potent risk factor for depression, yet the underlying mechanism is not clearly understood. In the present study, we explored the mechanism of development and maintenance of depression in a stress-induced animal model. Mice restrained for 2 h daily for 14 d showed distinct depressive behavior, and the altered behavior persisted for >3 months in the absence of intervention. Acute restraint induced a surge of oxidative stress in the brain, and stress-induced oxidative stress progressively increased with repetition of stress. In vitro, the stress hormone glucocorticoid generated superoxide via upregulation of NADPH oxidase. Consistently, repeated restraints increased the expression of the key subunits of NADPH oxidase, p47phox and p67phox, in the brain. Moreover, stressed brains markedly upregulated the expression of p47phox to weak restress evoked in the poststress period, and this molecular response was reminiscent of amplified ROS surge to restress. Pharmacological inhibition of NADPH oxidase by the NADPH oxidase inhibitor apocynin during the stress or poststress period completely blocked depressive behavior. Consistently, heterozygous p47phox knock-out mice (p47phox(+/-)) or molecular inhibition of p47phox with Lenti shRNA-p47phox in the hippocampus suppressed depressive behavior. These results suggest that repeated stress promotes depressive behavior through the upregulation of NADPH oxidase and the resultant metabolic oxidative stress, and that the inhibition of NADPH oxidase provides beneficial antidepression effects.

  1. Direct Identification of a Bacterial Manganese(II) Oxidase, the Multicopper Oxidase MnxG, from Spores of Several Different Marine Bacillus Species▿ †

    PubMed Central

    Dick, Gregory J.; Torpey, Justin W.; Beveridge, Terry J.; Tebo, Bradley M.

    2008-01-01

    Microorganisms catalyze the formation of naturally occurring Mn oxides, but little is known about the biochemical mechanisms of this important biogeochemical process. We used tandem mass spectrometry to directly analyze the Mn(II)-oxidizing enzyme from marine Bacillus spores, identified as an Mn oxide band with an in-gel activity assay. Nine distinct peptides recovered from the Mn oxide band of two Bacillus species were unique to the multicopper oxidase MnxG, and one peptide was from the small hydrophobic protein MnxF. No other proteins were detected in the Mn oxide band, indicating that MnxG (or a MnxF/G complex) directly catalyzes biogenic Mn oxide formation. The Mn(II) oxidase was partially purified and found to be resistant to many proteases and active even at high concentrations of sodium dodecyl sulfate. Comparative analysis of the genes involved in Mn(II) oxidation from three diverse Bacillus species revealed a complement of conserved Cu-binding regions not present in well-characterized multicopper oxidases. Our results provide the first direct identification of a bacterial enzyme that catalyzes Mn(II) oxidation and suggest that MnxG catalyzes two sequential one-electron oxidations from Mn(II) to Mn(III) and from Mn(III) to Mn(IV), a novel type of reaction for a multicopper oxidase. PMID:18165363

  2. The voltage dependence of NADPH oxidase reveals why phagocytes need proton channels

    NASA Astrophysics Data System (ADS)

    DeCoursey, Thomas E.; Morgan, Deri; Cherny, Vladimir V.

    2003-04-01

    The enzyme NADPH oxidase in phagocytes is important in the body's defence against microbes: it produces superoxide anions (O2-, precursors to bactericidal reactive oxygen species). Electrons move from intracellular NADPH, across a chain comprising FAD (flavin adenine dinucleotide) and two haems, to reduce extracellular O2 to O2-. NADPH oxidase is electrogenic, generating electron current (Ie) that is measurable under voltage-clamp conditions. Here we report the complete current-voltage relationship of NADPH oxidase, the first such measurement of a plasma membrane electron transporter. We find that Ie is voltage-independent from -100mV to >0mV, but is steeply inhibited by further depolarization, and is abolished at about +190mV. It was proposed that H+ efflux mediated by voltage-gated proton channels compensates Ie, because Zn2+ and Cd2+ inhibit both H+ currents and O2- production. Here we show that COS-7 cells transfected with four NADPH oxidase components, but lacking H+ channels, produce O2- in the presence of Zn2+ concentrations that inhibit O2- production in neutrophils and eosinophils. Zn2+ does not inhibit NADPH oxidase directly, but through effects on H+ channels. H+ channels optimize NADPH oxidase function by preventing membrane depolarization to inhibitory voltages.

  3. Nucleic Acid Homologies Among Oxidase-Negative Moraxella Species

    PubMed Central

    Johnson, John L.; Anderson, Robert S.; Ordal, Erling J.

    1970-01-01

    The deoxyribonucleic acid (DNA) base composition and DNA homologies of more than 40 strains of oxidase-negative Moraxella species were determined. These bacteria have also been identified as belonging to the Mima-Herellea-Acinetobacter group and the Bacterium anitratum group, as well as to several other genera including Achromobacter and Alcaligenes. The DNA base content of these strains ranged from 40 to 46% guanine plus cytosine. DNA–DNA competition experiments distinguished five groups whose members were determined by showing 50% or more homology to one of the reference strains: B. anitratum type B5W, Achromobacter haemolyticus var. haemolyticus, Alcaligenes haemolysans, Achromobacter metalcaligenes, and Moraxella lwoffi. A sixth group comprised those strains showing less than 50% homology to any of the reference strains. Negligible homology was found between strains of oxidase-negative and oxidase-positive Moraxella species in DNA–DNA competition experiments. However, evidence of a distant relationship between the two groups was obtained in competition experiments by using ribosomal ribonucleic acid. PMID:5413826

  4. [Activation of the alternative oxidase of Yarrowia lipolytica by adenosine 5'-monophosphate].

    PubMed

    Medentsev, A G; Arinbasarova, A Iu; Smirnova, N M; Akimenko, V K

    2004-01-01

    The study of the effect of nucleoside phosphates on the activity of cyanide-resistant oxidase in the mitochondria and the submitochondrial particles of Yarrowia lipolytica showed that adenosine monophosphate (5'-AMP, AMP) did not stimulate the respiration of the intact mitochondria. The incubation of the mitochondria at room temperature (25 degrees C) for 3-5 h or their treatment with ultrasound, phospholipase A, and detergent Triton X-100 at a low temperature inactivated the cyanide-resistant alternative oxidase. The inactivated alternative oxidase could be reactivated by AMP. The reactivating effect of AMP was enhanced by azolectin. Some other nucleoside phosphates also showed reactivating ability in the following descending order. AMP = GMP > GDP > GTP > XMP > IMP. The apparent reaction rate constant Km for AMP upon the reactivation of the alternative oxidase of mitochondria treated with Triton X-100 or incubated at 25 degrees C was 12.5 and 20 microM, respectively. The Km for AMP upon the reactivation of the alternative oxidase of submitochondrial particles was 15 microM. During the incubation of yeast cells under conditions promoting the development of alternative oxidase, the content of adenine nucleotides (AMP, ADP, and ATP) in the cells and their respiration tended to decrease. The subsequent addition of cyanide to the cells activated their respiration, diminished the intracellular content of ATP three times, and augmented the content of AMP five times. These data suggest that the stimulation of cell respiration by cyanide may be due to the activation of alternative oxidase by AMP.

  5. A randomized trial of assertive continuing care and contingency management for adolescents with substance use disorders.

    PubMed

    Godley, Mark D; Godley, Susan H; Dennis, Michael L; Funk, Rodney R; Passetti, Lora L; Petry, Nancy M

    2014-02-01

    Most adolescents relapse within 90 days of discharge from residential substance use treatment. We hypothesized that contingency management (CM), assertive continuing care (ACC), and their combination (CM + ACC) would each be more effective than usual continuing care (UCC). Following residential treatment, 337 adolescents were randomized to 4 continuing care conditions: UCC alone, CM, ACC, or CM + ACC. UCC was available across all conditions. Outcome measures over 12 months included percentage of days abstinent from alcohol, heavy alcohol, marijuana, and any alcohol or other drugs (AOD) using self-reports and toxicology testing and remission status at 12 months. CM had significantly higher rates of abstinence than UCC for heavy alcohol use, t(297) = 2.50, p < .01, d = 0.34; any alcohol use, t(297) = 2.58, p < .01, d = 0.36; or any AOD use, t(297) = 2.12, p = .01, d = 0.41; and had a higher rate in remission, odds ratio (OR) = 2.45, 90% confidence interval (CI) [1.18, 5.08], p = .02. ACC had significantly higher rates of abstinence than UCC from heavy alcohol use, t(297) = 2.66, p < .01, d = 0.31; any alcohol use, t(297) = 2.63, p < .01, d = 0.30; any marijuana use, t(297) = 1.95, p = .02, d = 0.28; or any AOD use, t(297) = 1.88, p = .02, d = 0.30; and had higher rates in remission, OR = 2.31, 90% CI [1.10, 4.85], p = .03. The ACC + CM condition was not significantly different from UCC on any outcomes. CM and ACC are promising continuing care approaches after residential treatment. Future research should seek to further improve their effectiveness. PsycINFO Database Record (c) 2014 APA, all rights reserved.

  6. Comparative study of different alcohol sensors based on Screen-Printed Carbon Electrodes.

    PubMed

    Costa Rama, Estefanía; Biscay, Julien; González García, María Begoña; Julio Reviejo, A; Pingarrón Carrazón, José Manuel; Costa García, Agustín

    2012-05-30

    Different very simple single-use alcohol enzyme sensors were developed using alcohol oxidase (AOX) from three different yeast, Hansenula sp., Pichia pastoris and Candida boidinii, and employing three different commercial mediator-based Screen-Printed Carbon Electrodes as transducers. The mediators tested, Prussian Blue, Ferrocyanide and Co-phthalocyanine were included into the ink of the working electrode. The procedure to obtain these sensors consists of the immobilization of the enzyme on the electrode surface by adsorption. For the immobilization, an AOX solution is deposited on the working electrode and left until dried (1h) at room temperature. The best results were obtained with the biosensor using Screen-Printed Co-phthalocyanine/Carbon Electrode and AOX from Hansenula sp. The reduced cobalt-phthalocyanine form is amperometrically detected at +0.4V (vs. Ag pseudo reference electrode). This sensor shows good sensitivity (1211 nA mM(-1)), high precision (2.1% RSD value for the slope value of the calibration plot) and wide linear response (0.05-1.00 mM) for ethanol determination. The sensor provides also accurate results for ethanol quantification in alcoholic drinks. Copyright © 2012 Elsevier B.V. All rights reserved.

  7. Systemic Manifestations in Pyridox(am)ine 5'-Phosphate Oxidase Deficiency.

    PubMed

    Guerriero, Réjean M; Patel, Archana A; Walsh, Brian; Baumer, Fiona M; Shah, Ankoor S; Peters, Jurriaan M; Rodan, Lance H; Agrawal, Pankaj B; Pearl, Phillip L; Takeoka, Masanori

    2017-11-01

    Pyridoxine is converted to its biologically active form pyridoxal-5-phosphate (P5P) by the enzyme pyridox(am)ine 5'-phosphate oxidase and serves as a cofactor in nearly 200 reactions in the central nervous system. Pyridox(am)ine 5'-phosphate oxidase deficiency leads to P5P dependent epilepsy, typically a neonatal- or infantile-onset epileptic encephalopathy treatable with P5P or in some cases, pyridoxine. Following identification of retinopathy in a patient with pyridox(am)ine 5'-phosphate oxidase deficiency that was reversible with P5P therapy, we describe the systemic manifestations of pyridox(am)ine 5'-phosphate oxidase deficiency. A series of six patients with homozygous mutations of PNPO, the gene coding pyridox(am)ine 5'-phosphate oxidase, were evaluated in our center over the course of two years for phenotyping of neurological and systemic manifestations. Five of six were born prematurely, three had anemia and failure to thrive, and two had elevated alkaline phosphatase. A movement disorder was observed in two children, and a reversible retinopathy was observed in the most severely affected infant. All patients had neonatal-onset epilepsy and were on a continuum of developmental delay to profound encephalopathy. Electroencephalographic features included background slowing and disorganization, absent sleep features, and multifocal and generalized epileptiform discharges. All the affected probands carried a homozygous PNPO mutation (c.674 G>T, c.686 G>A and c.352G>A). In addition to the well-described epileptic encephalopathy, pyridox(am)ine 5'-phosphate oxidase deficiency causes a range of neurological and systemic manifestations. A movement disorder, developmental delay, and encephalopathy, as well as retinopathy, anemia, and failure to thrive add to the broadening clinical spectrum of P5P dependent epilepsy. Copyright © 2017 Elsevier Inc. All rights reserved.

  8. Xanthine Oxidase Induces Foam Cell Formation through LOX-1 and NLRP3 Activation.

    PubMed

    Dai, Yao; Cao, Yongxiang; Zhang, Zhigao; Vallurupalli, Srikanth; Mehta, Jawahar L

    2017-02-01

    Xanthine oxidase catalyzes the oxidation of xanthine to uric acid. This process generates excessive reactive oxygen species (ROS) that play an important role in atherogenesis. Recent studies show that LRR and PYD domains-containing protein 3 (NLRP3), a component of the inflammasome, may be involved in the formation of foam cells, a hallmark of atherosclerosis. This study was designed to study the role of various scavenger receptors and NLRP3 inflammasome in xanthine oxidase and uric acid-induced foam cell formation. Human vascular smooth muscle cells (VSMCs) and THP-1 macrophages were treated with xanthine oxidase or uric acid. Xanthine oxidase treatment (of both VSMCs and THP-1 cells) resulted in foam cell formation in concert with generation of ROS and expression of cluster of differentiation 36 (CD36) and oxidized low density lipoprotein (lectin-like) receptor 1 (LOX-1), but not of scavenger receptor A (SRA). Uric acid treatment resulted in foam cell formation, ROS generation and expression of CD36, but not of LOX-1 or SRA. Further, treatment of cells with xanthine oxidase, but not uric acid, activated NLRP3 and its downstream pro-inflammatory signals- caspase-1, interleukin (IL)-1β and IL-18. Blockade of LOX-1 or NLRP3 inflammasome with specific siRNAs reduced xanthine oxidase-induced foam cell formation, ROS generation and activation of NLRP3 and downstream signals. Xanthine oxidase induces foam cell formation in large part through activation of LOX-1 - NLRP3 pathway in both VSMCs and THP-1 cells, but uric acid-induced foam cell formation is exclusively through CD36 pathway. Further, LOX-1 activation is upstream of NLRP3 activation. Graphical Abstract Steps in the formation of foam cells in response to xanthine oxidase and uric acid. Xanthine oxidase stimulates LOX-1 expression on the cell membrane of macrophages and vascular smooth muscle cells (VSMCs) and increases generation of ROS, which activate NLRP3 inflammasome and downstream pro

  9. Parkin regulates mitophagy and mitochondrial function to protect against alcohol-induced liver injury and steatosis in mice

    PubMed Central

    Williams, Jessica A.; Ni, Hong-Min; Ding, Yifeng

    2015-01-01

    Alcoholic liver disease claims two million lives per year. We previously reported that autophagy protected against alcohol-induced liver injury and steatosis by removing damaged mitochondria. However, the mechanisms for removal of these mitochondria are unknown. Parkin is an evolutionarily conserved E3 ligase that is recruited to damaged mitochondria to initiate ubiquitination of mitochondrial outer membrane proteins and subsequent mitochondrial degradation by mitophagy. In addition to its role in mitophagy, Parkin has been shown to have other roles in maintaining mitochondrial function. We investigated whether Parkin protected against alcohol-induced liver injury and steatosis using wild-type (WT) and Parkin knockout (KO) mice treated with alcohol by the acute-binge and Gao-binge (chronic plus acute-binge) models. We found that Parkin protected against liver injury in both alcohol models, likely because of Parkin's role in maintaining a population of healthy mitochondria. Alcohol caused greater mitochondrial damage and oxidative stress in Parkin KO livers compared with WT livers. After alcohol treatment, Parkin KO mice had severely swollen and damaged mitochondria that lacked cristae, which were not seen in WT mice. Furthermore, Parkin KO mice had decreased mitophagy, β-oxidation, mitochondrial respiration, and cytochrome c oxidase activity after acute alcohol treatment compared with WT mice. Interestingly, liver mitochondria seemed able to adapt to alcohol treatment, but Parkin KO mouse liver mitochondria had less capacity to adapt to Gao-binge treatment compared with WT mouse liver mitochondria. Overall, our findings indicate that Parkin is an important mediator of protection against alcohol-induced mitochondrial damage, steatosis, and liver injury. PMID:26159696

  10. Protein structural development of threadfin bream ( Nemipterus spp.) surimi gels induced by glucose oxidase.

    PubMed

    Wang, Lei; Fan, Daming; Fu, Lulu; Jiao, Xidong; Huang, Jianlian; Zhao, Jianxin; Yan, Bowen; Zhou, Wenguo; Zhang, Wenhai; Ye, Weijian; Zhang, Hao

    2018-01-01

    This study investigated the effect of glucose oxidase on the gel properties of threadfin bream surimi. The gel strength of surimi increased with the addition of 0.5‰ glucose oxidase after two-step heating. Based on the results of the chemical interactions, the hydrophobic interaction and disulfide bond of glucose oxidase-treated surimi samples increased compared with the control samples at the gelation temperature and gel modori temperature. The surface hydrophobicity of samples with glucose oxidase and glucose increased significantly ( p < 0.05) and total sulfhydryl groups decreased significantly ( p < 0.05). The analysis of Raman spectroscopy shows that the addition of glucose oxidase induced more α-helixes to turn into a more elongated random and flocculent structure. Glucose oxidase changes the secondary structure of the surimi protein, making more proteins depolarize and stretch and causing actomyosin to accumulate to each other, resulting in the formation of surimi gel.

  11. Altered brain functional connectivity and behaviour in a mouse model of maternal alcohol binge-drinking.

    PubMed

    Cantacorps, Lídia; González-Pardo, Héctor; Arias, Jorge L; Valverde, Olga; Conejo, Nélida M

    2018-06-08

    Prenatal and perinatal alcohol exposure caused by maternal alcohol intake during gestation and lactation periods can have long-lasting detrimental effects on the brain development and behaviour of offspring. Children diagnosed with Foetal Alcohol Spectrum Disorders (FASD) display a wide range of cognitive, emotional and motor deficits, together with characteristic morphological abnormalities. Maternal alcohol binge drinking is particularly harmful for foetal and early postnatal brain development, as it involves exposure to high levels of alcohol over short periods of time. However, little is known about the long-term effects of maternal alcohol binge drinking on brain function and behaviour. To address this issue, we used pregnant C57BL/6 female mice with time-limited access to a 20% v/v alcohol solution as a procedure to model alcohol binge drinking during gestation and lactational periods. Male offspring were behaviourally tested during adolescence (30 days) and adulthood (60 days), and baseline neural metabolic capacity of brain regions sensitive to alcohol effects were also evaluated in adult animals from both groups. Our results show that prenatal and postnatal alcohol exposure caused age-dependent changes in spontaneous locomotor activity, increased anxiety-like behaviour and attenuated alcohol-induced conditioned place preference in adults. Also, significant changes in neural metabolic capacity using cytochrome c oxidase (CCO) quantitative histochemistry were found in the hippocampal dentate gyrus, the mammillary bodies, the ventral tegmental area, the lateral habenula and the central lobules of the cerebellum in adult mice with prenatal and postnatal alcohol exposure. In addition, the analysis of interregional CCO activity correlations in alcohol-exposed adult mice showed disrupted functional brain connectivity involving the limbic, brainstem, and cerebellar regions. Finally, increased neurogenesis was found in the dentate gyrus of the hippocampus of

  12. NADPH Oxidase as a Therapeutic Target for Oxalate Induced Injury in Kidneys

    PubMed Central

    Peck, Ammon B.; Khan, Saeed R.

    2013-01-01

    A major role of the nicotinamide adenine dinucleotide phosphate (NADPH) oxidase family of enzymes is to catalyze the production of superoxides and other reactive oxygen species (ROS). These ROS, in turn, play a key role as messengers in cell signal transduction and cell cycling, but when they are produced in excess they can lead to oxidative stress (OS). Oxidative stress in the kidneys is now considered a major cause of renal injury and inflammation, giving rise to a variety of pathological disorders. In this review, we discuss the putative role of oxalate in producing oxidative stress via the production of reactive oxygen species by isoforms of NADPH oxidases expressed in different cellular locations of the kidneys. Most renal cells produce ROS, and recent data indicate a direct correlation between upregulated gene expressions of NADPH oxidase, ROS, and inflammation. Renal tissue expression of multiple NADPH oxidase isoforms most likely will impact the future use of different antioxidants and NADPH oxidase inhibitors to minimize OS and renal tissue injury in hyperoxaluria-induced kidney stone disease. PMID:23840917

  13. [Experimental rationale for the parameters of a rapid method for oxidase activity determination].

    PubMed

    Butorina, N N

    2010-01-01

    Experimental rationale is provided for the parameters of a rapid (1-2-min) test to concurrently determine the oxidase activity of all bacteria grown on the membrane filter after water filtration. Oxidase reagents that are the aqueous solutions of tetramethyl-p-phenylenediamine dihydrochloride and demethyl-p-phenylenediamine dihydrochloride have been first ascertained to exert no effect on the viability and enzymatic activity of bacteria after one-hour contact. An algorithm has been improved for the rapid oxidase activity test: the allowable time for bacteria to contact oxidase reagents and procedures for minimizing the effect on bacterial biochemical activity following the contact. An accelerated method based on lactose medium with tergitol 7 and Endo agar has been devised to determine coliform bacteria, by applying the rapid oxidase test: the time of a final response is 18-24 hours. The method has been included into GOST 52426-2005.

  14. Heterologous production and characterization of two glyoxal oxidases from Pycnoporus cinnabarinus

    Treesearch

    Marianne Daou; François Piumi; Daniel Cullen; Eric Record; Craig B. Faulds

    2016-01-01

    The genome of the white rot fungus Pycnoporus cinnabarinus includes a large number of genes encoding enzymes implicated in lignin degradation. Among these, three genes are predicted to encode glyoxal oxidase, an enzyme previously isolated from Phanerochaete chrysosporium. The glyoxal oxidase of P. chrysosporium...

  15. Phospholipid alterations in cardiac sarcoplasmic reticulum induced by xanthine oxidase: contamination of commercial preparations of xanthine oxidase by phospholipase A/sub 2/

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

    Gamache, D.A.; Kornberg, L.J.; Bartolf, M.

    1986-05-01

    Incubation of cardiac sarcoplasmic reticulum with xanthine oxidase alone at pH 7.0 resulted in a loss of lipid phosphorus that was potentiated by the addition of xanthine. Using autoclaved E.coli with 1-/sup 14/C-oleate in the 2-acyl position of membrane phospholipids, the authors demonstrate that many, but not all, commercial preparations of xanthine oxidase contain significant phospholipase A/sub 2/ (PLA/sub 2/) activity (64.3-545.6 nmols/min/mg). The PLA/sub 2/ was maximally active in the neutral-alkaline pH range, was Ca/sup 2 +/-dependent, and was unaffected by the addition of xanthine. PLA/sub 2/ activity was totally inhibited by 1mM EDTA whereas radical production by optimalmore » concentrations of xanthine/xanthine oxidase (X/XO) was unaffected by EDTA. Chromatographically purified xanthine oxidase (Sigma Grade III) contained high levels of PLA/sub 2/ activity (64.3 nmols/min/mg) compared to endogenous levels of neutral-active, Ca/sup 2 +/-dependent PLA/sub 2/ measured in various tissue homogenates (less than or equal to 0.5 nmols/ min/mg). Because X/XO mixtures are used extensively to study oxygen free radical-induced cell injury and membrane phospholipid alterations, the presence of a potent extracellular PLA/sub 2/ may have influenced previously published reports, and such studies should be interpreted cautiously.« less

  16. A Biochemical Approach to Study the Role of the Terminal Oxidases in Aerobic Respiration in Shewanella oneidensis MR-1

    PubMed Central

    Le Laz, Sébastien; Kpebe, Arlette; Bauzan, Marielle; Lignon, Sabrina; Rousset, Marc; Brugna, Myriam

    2014-01-01

    The genome of the facultative anaerobic γ-proteobacterium Shewanella oneidensis MR-1 encodes for three terminal oxidases: a bd-type quinol oxidase and two heme-copper oxidases, a A-type cytochrome c oxidase and a cbb 3-type oxidase. In this study, we used a biochemical approach and directly measured oxidase activities coupled to mass-spectrometry analysis to investigate the physiological role of the three terminal oxidases under aerobic and microaerobic conditions. Our data revealed that the cbb 3-type oxidase is the major terminal oxidase under aerobic conditions while both cbb 3-type and bd-type oxidases are involved in respiration at low-O2 tensions. On the contrary, the low O2-affinity A-type cytochrome c oxidase was not detected in our experimental conditions even under aerobic conditions and would therefore not be required for aerobic respiration in S. oneidensis MR-1. In addition, the deduced amino acid sequence suggests that the A-type cytochrome c oxidase is a ccaa 3-type oxidase since an uncommon extra-C terminal domain contains two c-type heme binding motifs. The particularity of the aerobic respiratory pathway and the physiological implication of the presence of a ccaa 3-type oxidase in S. oneidensis MR-1 are discussed. PMID:24466040

  17. Phenol oxidase activity in secondary transformed peat-moorsh soils

    NASA Astrophysics Data System (ADS)

    Styła, K.; Szajdak, L.

    2009-04-01

    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

  18. Design, synthesis and molecular modeling of aloe-emodin derivatives as potent xanthine oxidase inhibitors.

    PubMed

    Shi, Da-Hua; Huang, Wei; Li, Chao; Liu, Yu-Wei; Wang, Shi-Fan

    2014-03-21

    A series of aloe-emodin derivatives were synthesized and evaluated as xanthine oxidase inhibitors. Among them, four aloe-emodin derivatives showed significant inhibitory activities against xanthine oxidase. The compound 4,5-dihydroxy-9,10-dioxo-9,10-dihydroanthracene-2-carbaldehyde (A1) possessed the best xanthine oxidase inhibitory activity with IC50 of 2.79 μM. Lineweaver-Burk plot analysis revealed that A1 acted as a mixed-type inhibitor for xanthine oxidase. The docking study revealed that the molecule A1 had strong interactions with the active site of xanthine oxidase and this result was in agreement with kinetic study. Consequently, compound A1 is a new-type candidate for further development for the treatment of gout. Copyright © 2014 Elsevier Masson SAS. All rights reserved.

  19. Youth and Drugs: Society's Mixed Messages. OSAP Prevention Monograph-6.

    ERIC Educational Resources Information Center

    Resnik, Hank; And Others

    This document examines societal influences that promote the continued use of drugs, considering how "mixed messages" are sent to U.S. youth regarding the acceptable use of alcohol and other drugs (AOD). It analyzes public and private sector policies, practices, and regulatory initiatives aimed at reducing and eliminating alcohol and other drug…

  20. Supplementary biochemical tests useful for the differentiation of oxidase positive staphylococci.

    PubMed

    Stepanović, Srdjan; Dakić, Ivana; Hauschild, Tomasz; Vuković, Dragana; Morrison, Donald; Jezek, Petr; Cirković, Ivana; Petrás, Petr

    2007-06-01

    Differentiation of the oxidase positive staphylococci, Staphylococcus sciuri, Staphylococcus lentus, Staphylococcus vitulinus and Staphylococcus fleurettii, based on tributyrin, urease, caseinase, gelatinase and DNase activity is described. These tests may be used for preliminary identification of oxidase positive isolates of staphylococci resulting in more accurate identification of these species.

  1. NADPH oxidases: new kids on the block.

    PubMed

    Geiszt, Miklós

    2006-07-15

    Reactive oxygen species (ROS) play a pivotal role in many physiological processes including host defense, hormone biosynthesis, fertilization and cellular signaling. Altered production of ROS has been implicated in the development of immunodeficiency, hypothyroidism and cardiovascular pathologies. In the last few years, several enzymes were identified at the molecular level, which are now thought to be responsible for ROS production observed in diverse tissues. These enzymes show a high degree of homology to the phagocytic NADPH oxidase and are now designated the Nox family of NADPH oxidases. This review updates our knowledge on six new members of the Nox family: Nox1, Nox3, Nox4, Nox5, Duox1 and Duox2.

  2. Monocyte and macrophage-targeted NADPH oxidase mediates antifungal host defense and regulation of acute inflammation in mice

    PubMed Central

    Grimm, Melissa J.; Vethanayagam, R. Robert; Almyroudis, Nikolaos G.; Dennis, Carly G.; Khan, A. Nazmul H.; D’Auria, Anthony; Singel, Kelly L.; Davidson, Bruce A.; Knight, Paul R.; Blackwell, Timothy S.; Hohl, Tobias M.; Mansour, Michael K.; Vyas, Jatin M.; Röhm, Marc; Urban, Constantin F.; Kelkka, Tiina; Holmdahl, Rikard; Segal, Brahm H.

    2013-01-01

    Chronic granulomatous disease, an inherited disorder of the NADPH oxidase in which phagocytes are defective in the generation of superoxide anion and downstream reactive oxidant species, is characterized by severe bacterial and fungal infections and excessive inflammation. Although NADPH oxidase isoforms exist in several lineages, reactive oxidant generation is greatest in neutrophils, where NADPH oxidase has been deemed vital for pathogen killing. In contrast, the function and importance of NADPH oxidase in macrophages are less clear. Therefore, we evaluated susceptibility to pulmonary aspergillosis in globally NADPH oxidase-deficient mice versus transgenic mice with monocyte/macrophage-targeted NADPH oxidase activity. We found that the lethal inoculum was more than 100-fold greater in transgenic versus globally NADPH oxidase-deficient mice. Consistent with these in vivo results, NADPH oxidase in mouse alveolar macrophages limited germination of phagocytosed Aspergillus fumigatus spores. Finally, globally NADPH oxidase-deficient mice developed exuberant neutrophilic lung inflammation and pro-inflammatory cytokine responses to zymosan, a fungal cell wall-derived product composed principally of particulate beta-glucans, whereas inflammation in transgenic and wildtype mice was mild and transient. Together, our studies identify a central role for monocyte/macrophage NADPH oxidase in controlling fungal infection and in limiting acute lung inflammation. PMID:23509361

  3. Campus and Community Coalitions in AOD Prevention. Prevention Updates

    ERIC Educational Resources Information Center

    Zimmerman, Robert

    2004-01-01

    Changing the environment that affects decisions about use of alcohol and other drugs has emerged as a key to reducing substance use problems among college and university students. The student environment, however, is not limited to the campus. Education, counseling, and campus-based prevention policies and activities cannot succeed if a campus is…

  4. Effect of mitoguazone on polyamine oxidase activity in rat liver.

    PubMed

    Ferioli, Maria Elena; Berselli, Debora; Caimi, Samuela

    2004-12-01

    Mitoguazone is a known inhibitor of polyamine biosynthesis through competitive inhibition of S-adenosylmethionine decarboxylase. A recent renewed interest in mitoguazone as an antineoplastic agent prompted us to investigate the effect of the drug on polyamine catabolism in rat liver, since the organ plays an important role in detoxification mechanisms. Thus, the purpose of this work was to evaluate the effect of in vivo mitoguazone administration on polyamine catabolic enzymes. In particular, our interest was directed to the changes in polyamine oxidase activity, since this enzyme has been recently confirmed to exert important functions that until now were underestimated. Mitoguazone administration induced hepatic polyamine oxidase activity starting at 4 h after administration, and the enzyme returned to basal levels 96 h after treatment. The changes in enzyme activity were accompanied by changes in putrescine concentrations, which increased starting at 4 h until 72 h after treatment. We also evaluated the activity of the newly identified spermine oxidase, which was not significantly changed by mitoguazone treatment. Therefore, we hypothesized that the enzyme involved in mitoguazone response of the liver is the polyamine oxidase, which acts on acetylated polyamines as substrate.

  5. Oxidase-functionalized Fe(3)O(4) nanoparticles for fluorescence sensing of specific substrate.

    PubMed

    Liu, Cheng-Hao; Tseng, Wei-Lung

    2011-10-03

    This study reports the development of a reusable, single-step system for the detection of specific substrates using oxidase-functionalized Fe(3)O(4) nanoparticles (NPs) as a bienzyme system and using amplex ultrared (AU) as a fluorogenic substrate. In the presence of H(2)O(2), the reaction pH between Fe(3)O(4) NPs and AU was similar to the reaction of oxidase and the substrate. The catalytic activity of Fe(3)O(4) NPs with AU was nearly unchanged following modification with poly(diallyldimethylammonium chloride) (PDDA). Based on these features, we prepared a composite of PDDA-modified Fe(3)O(4) NPs and oxidase for the quantification of specific substrates through the H(2)O(2)-mediated oxidation of AU. By monitoring fluorescence intensity at 587 nm of oxidized AU, the minimum detectable concentrations of glucose, galactose, and choline were found to be 3, 2, and 20 μM using glucose oxidase-Fe(3)O(4), galactose oxidase-Fe(3)O(4), and choline oxidase-Fe(3)O(4) composites, respectively. The identification of glucose in blood was selected as the model to validate the applicability of this proposed method. Copyright © 2011 Elsevier B.V. All rights reserved.

  6. A Mycobacterium tuberculosis Cytochrome bd Oxidase Mutant Is Hypersensitive to Bedaquiline

    PubMed Central

    Hartman, Travis E.

    2014-01-01

    ABSTRACT The new medicinal compound bedaquiline (BDQ) kills Mycobacterium tuberculosis by inhibiting F1Fo-ATP synthase. BDQ is bacteriostatic for 4 to 7 days and kills relatively slowly compared to other frontline tuberculosis (TB) drugs. Here we show that killing with BDQ can be improved significantly by inhibiting cytochrome bd oxidase, a non-proton-pumping terminal oxidase. BDQ was instantly bactericidal against a cytochrome bd oxidase null mutant of M. tuberculosis, and the rate of killing was increased by more than 50%. We propose that this exclusively bacterial enzyme should be a high-priority target for new drug discovery. PMID:25028424

  7. Plasma amine oxidase activities in Norrie disease patients with an X-chromosomal deletion affecting monoamine oxidase.

    PubMed

    Murphy, D L; Sims, K B; Karoum, F; Garrick, N A; de la Chapelle, A; Sankila, E M; Norio, R; Breakefield, X O

    1991-01-01

    Two individuals with an X-chromosomal deletion were recently found to lack the genes encoding monoamine oxidase type A (MAO-A) and MAO-B. This abnormality was associated with almost total (90%) reductions in the oxidatively deaminated urinary metabolites of the MAO-A substrate, norepinephrine, and with marked (100-fold) increases in an MAO-B substrate, phenylethylamine, confirming systemic functional consequences of the genetic enzyme deficiency. However, urinary concentrations of the deaminated metabolites of dopamine and serotonin (5-HT) were essentially normal. To investigate other deaminating systems besides MAO-A and MAO-B that might produce these metabolites of dopamine and 5-HT, we examined plasma amine oxidase (AO) activity in these two patients and two additional patients with the same X-chromosomal deletion. Normal plasma AO activity was found in all four Norrie disease-deletion patients, in four patients with classic Norrie disease without a chromosomal deletion, and in family members of patients from both groups. Marked plasma amine metabolite abnormalities and essentially absent platelet MAO-B activity were found in all four Norrie disease-deletion patients, but in none of the other subjects in the two comparison groups. These results indicate that plasma AO is encoded by gene(s) independent of those for MAO-A and MAO-B, and raise the possibility that plasma AO, and perhaps the closely related tissue AO, benzylamine oxidase, as well as other atypical AOs or MAOs encoded independently from MAO-A and MAO-B may contribute to the oxidative deamination of dopamine and 5-HT in humans.

  8. NADPH oxidase activation in neutrophils: Role of the Phosphorylation of its subunits.

    PubMed

    Belambri, Sahra A; Rolas, Loïc; Raad, Houssam; Hurtado-Nedelec, Margarita; Dang, Pham My-Chan; El-Benna, Jamel

    2018-05-14

    Neutrophils are key cells of innate immunity and during inflammation. Upon activation, they produce large amounts of superoxide anion (O 2 -. ) and ensuing reactive oxygen species (ROS) to kill phagocytized microbes. The enzyme responsible for O 2 -. production is called the phagocyte NADPH oxidase. This is a multicomponent enzyme system that becomes active after assembly of four cytosolic proteins (p47 phox , p67 phox , p40 phox and Rac2) with the transmembrane proteins (p22 phox and gp91 phox , which form the cytochrome b 558 ). gp91 phox represents the catalytic subunit of the NADPH oxidase and is also called NOX2. NADPH oxidase-derived ROS are essential for microbial killing and innate immunity; however, excessive ROS production induces tissue injury and prolonged inflammatory reactions that contribute to inflammatory diseases. Thus, NADPH oxidase activation must be tightly regulated in time and space in order to limit ROS production. NADPH oxidase activation is regulated by several processes such as phosphorylation of its components, exchange of GDP/GTP on Rac2 and binding of p47 phox and p40 phox to phospholipids. This review aims to provide new insights into the role of the phosphorylation of the NADPH oxidase components, i.e., gp91 phox , p22 phox , p47 phox , p67 phox and p40 phox , in the activation of this enzyme. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

  9. Substitutions of S101 decrease proton and hydride transfers in the oxidation of betaine aldehyde by choline oxidase.

    PubMed

    Gadda, Giovanni; Yuan, Hongling

    2017-11-15

    Choline oxidase oxidizes choline to glycine betaine, with two flavin-mediated reactions to convert the alcohol substrate to the carbon acid product. Proton abstraction from choline or hydrated betaine aldehyde in the wild-type enzyme occurs in the mixing time of the stopped-flow spectrophotometer, thereby precluding a mechanistic investigation. Mutagenesis of S101 rendered the proton transfer reaction amenable to study. Here, we have investigated the aldehyde oxidation reaction catalyzed by the mutant enzymes using steady-state and rapid kinetics with betaine aldehyde. Stopped-flow traces for the reductive half-reaction of the S101T/V/C variants were biphasic, corresponding to the reactions of proton abstraction and hydride transfer. In contrast, the S101A enzyme yielded monophasic traces like wild-type choline oxidase. The rate constants for proton transfer in the S101T/C/V variants decreased logarithmically with increasing hydrophobicity of residue 101, indicating a behavior different from that seen previously with choline for which no correlation was determined. The rate constants for hydride transfer also showed a logarithmic decrease with increasing hydrophobicity at position 101, which was similar to previous results with choline as a substrate for the enzyme. Thus, the hydrophilic character of S101 is necessary not only for efficient hydride transfer but also for the proton abstraction reaction. Copyright © 2017. Published by Elsevier Inc.

  10. Safety assessment of bacterial choline oxidase protein introduced in transgenic crops for tolerance against abiotic stress.

    PubMed

    Singh, Abinav K; Singh, Bhanu P; Prasad, G B K S; Gaur, Shailendra N; Arora, Naveen

    2008-12-24

    Genetically modified crops have resistance to abiotic stress by introduction of choline oxidase protein. In the present study, the safety of choline oxidase protein derived from Arthrobacter globiformis was assessed for toxicity and allergenicity. The protein was stable at 90 degrees C for 1 h. Toxicity studies of choline oxidase in mice showed no significant difference (p > 0.05) from control in terms of growth, body weight, food consumption, and blood biochemical indices. Histology of gut tissue of mice fed protein showed normal gastric mucosal lining and villi in jejunum and ileum sections. Specific IgE in serum and IL-4 release in splenic culture supernatant were low in choline oxidase treated mice, comparable to control. Intravenous challenge with choline oxidase did not induce any adverse reaction, unlike ovalbumin group mice. Histology of lung tissues from choline oxidase sensitized mice showed normal airways, whereas ovalbumin-sensitized mice showed inflamed airways with eosinophilic infiltration and bronchoconstriction. ELISA carried out with food allergic patients' sera revealed no significant IgE affinity with choline oxidase. Also, choline oxidase did not show any symptoms of toxicity and allergenicity in mice.

  11. Inactivation of 1-aminocyclopropane-1-carboxylate oxidase involves oxidative modifications.

    PubMed

    Barlow, J N; Zhang, Z; John, P; Baldwin, J E; Schofield, C J

    1997-03-25

    1-Aminocyclopropane-1-carboxylate (ACC) oxidase catalyzes the final step in the biosynthesis of the plant signaling molecule ethylene. It is a member of the ferrous iron dependent family of oxidases and dioxygenases and is unusual in that it displays a very short half-life under catalytic conditions, typically less than 20 min, and a requirement for CO2 as an activator. The rates of inactivation of purified, recombinant ACC oxidase from tomato under various combinations of substrates and cofactors were measured. Inactivation was relatively slow in the presence of buffer alone (t1/2 > 1 h), but fast in the presence of ferrous iron and ascorbate (t1/2 approximately 10 min). The rate of iron/ascorbate-mediated inactivation was increased by the addition of ACC, unaffected by the addition of CO2 at saturation (supplied as bicarbonate) but decreased by the addition of catalase or ACC + CO2 at saturation (supplied as bicarbonate). Iron/ascorbate-mediated inactivation was accompanied by partial proteolysis as observed by SDS-PAGE analysis. The fragmentation pattern was altered when ACC was also included, suggesting that ACC can bind to ACC oxidase in the absence of bicarbonate. N-terminal sequencing of fragments resulted in identification of an internal cleavage site which we propose is proximate to active-site bound iron. Thus, ACC oxidase inactivates via relatively slow partial unfolding of the catalytically active conformation, oxidative damage mediated via hydrogen peroxide which is catalase protectable and oxidative damage to the active site which results in partial proteolysis and is not catalase protectable.

  12. NADPH oxidases as novel pharmacologic targets against influenza A virus infection.

    PubMed

    Vlahos, Ross; Selemidis, Stavros

    2014-12-01

    Influenza A viruses represent a major global health care challenge, with imminent pandemics, emerging antiviral resistance, and long lag times for vaccine development, raising a pressing need for novel pharmacologic strategies that ideally target the pathology irrespective of the infecting strain. Reactive oxygen species (ROS) pervade all facets of cell biology with both detrimental and protective properties. Indeed, there is compelling evidence that activation of the NADPH oxidase 2 (NOX2) isoform of the NADPH oxidase family of ROS-producing enzymes promotes lung oxidative stress, inflammation, injury, and dysfunction resulting from influenza A viruses of low to high pathogenicity, as well as impeding virus clearance. By contrast, the dual oxidase isoforms produce ROS that provide vital protective antiviral effects for the host. In this review, we propose that inhibitors of NOX2 are better alternatives than broad-spectrum antioxidant approaches for treatment of influenza pathologies, for which clinical efficacy may have been limited owing to poor bioavailability and inadvertent removal of beneficial ROS. Finally, we briefly describe the current suite of NADPH oxidase inhibitors and the molecular features of the NADPH oxidase enzymes that could be exploited by drug discovery for development of more specific and novel inhibitors to prevent or treat disease caused by influenza. Copyright © 2014 by The American Society for Pharmacology and Experimental Therapeutics.

  13. Parkin regulates mitophagy and mitochondrial function to protect against alcohol-induced liver injury and steatosis in mice.

    PubMed

    Williams, Jessica A; Ni, Hong-Min; Ding, Yifeng; Ding, Wen-Xing

    2015-09-01

    Alcoholic liver disease claims two million lives per year. We previously reported that autophagy protected against alcohol-induced liver injury and steatosis by removing damaged mitochondria. However, the mechanisms for removal of these mitochondria are unknown. Parkin is an evolutionarily conserved E3 ligase that is recruited to damaged mitochondria to initiate ubiquitination of mitochondrial outer membrane proteins and subsequent mitochondrial degradation by mitophagy. In addition to its role in mitophagy, Parkin has been shown to have other roles in maintaining mitochondrial function. We investigated whether Parkin protected against alcohol-induced liver injury and steatosis using wild-type (WT) and Parkin knockout (KO) mice treated with alcohol by the acute-binge and Gao-binge (chronic plus acute-binge) models. We found that Parkin protected against liver injury in both alcohol models, likely because of Parkin's role in maintaining a population of healthy mitochondria. Alcohol caused greater mitochondrial damage and oxidative stress in Parkin KO livers compared with WT livers. After alcohol treatment, Parkin KO mice had severely swollen and damaged mitochondria that lacked cristae, which were not seen in WT mice. Furthermore, Parkin KO mice had decreased mitophagy, β-oxidation, mitochondrial respiration, and cytochrome c oxidase activity after acute alcohol treatment compared with WT mice. Interestingly, liver mitochondria seemed able to adapt to alcohol treatment, but Parkin KO mouse liver mitochondria had less capacity to adapt to Gao-binge treatment compared with WT mouse liver mitochondria. Overall, our findings indicate that Parkin is an important mediator of protection against alcohol-induced mitochondrial damage, steatosis, and liver injury. Copyright © 2015 the American Physiological Society.

  14. To Listen and To Learn: Exploring Opportunities To Encourage Family Communication.

    ERIC Educational Resources Information Center

    Bankston, Karen, Ed.; Franklin, Diane, Ed.

    This first publication in the "Who's Raising the Family" series covers the critical issue of family communications and how parents and adolescents can talk and listen to each other about alcohol and other drugs (AOD). Communication between parents and children is crucial in preventing alcohol and other drug use. While peer relationships take on…

  15. Activation of monoamine oxidase isotypes by prolonged intake of aluminum in rat brain.

    PubMed

    Huh, Jae-Wan; Choi, Myung-Min; Lee, Jang Han; Yang, Seung-Ju; Kim, Mi Jung; Choi, Jene; Lee, Kwan Ho; Lee, Jong Eun; Cho, Sung-Woo

    2005-10-01

    Rats were fed 100 microM aluminum maltolate for one year in their drinking water. Brain aluminum contents have increased 4.2-fold in the aluminum-treated group, whereas no significant changes in the body weight, brain weight, and brain protein content were observed. Long-term aluminum feeding induced apoptosis as assessed by the terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling method and showed activatory effects on the catalytic efficiency (kcat/KM) of monoamine oxidase-A and monoamine oxidase-B up to 1.9- and 3.8-fold, respectively. The expression level of monoamine oxidase isotypes on the Western blot remained unchanged between the two groups, suggesting a change in post-translational regulation of the activities of monoamine oxidase isotypes by long-term aluminum feeding.

  16. A survey of 100 community colleges on student substance use, programming, and collaborations.

    PubMed

    Chiauzzi, Emil; Donovan, Elizabeth; Black, Ryan; Cooney, Elizabeth; Buechner, Allison; Wood, Mollie

    2011-01-01

    The objective was to survey community college personnel about student substance use, and infrastructure (staff and funding), programs, and collaborations dedicated to substance use prevention. The sample included 100 administrators, faculty, and health services staff at 100 community colleges. Participants completed a Web-based survey. Participants reported a number of alcohol and other drug (AOD) related concerns. Despite limited staff and funding dedicated to AOD, institutions are implementing a number of programs, although many are not implementing some of the programs popular at traditional 4-year colleges. They are also collaborating with a number of on- and off-campus groups. The availability of staff and funding dedicated to AOD, and the presence of residence halls, is associated with health programming and substance abuse collaborations. Results suggest that there is a need for increased research to understand the most effective AOD prevention strategies for community colleges.

  17. Purification, characterization and decolorization of bilirubin oxidase from Myrothecium verrucaria 3.2190

    USDA-ARS?s Scientific Manuscript database

    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...

  18. Towards rational therapy with monoamine oxidase inhibitors.

    PubMed

    Tyrer, P

    1976-04-01

    A rational approach to the use of monoamine oxidase inhibitors (MAOIs) is outlined. Patients suitable for treatment cannot be classified adequately using conventional diagnostic labels. They include those with primary symptoms of hypochondriasis, agoraphobia and social phobias, irritability, somatic anxiety and anergia; those with primary depressed mood, guilt, ideas of reference and personality disorders seldom respond. There is great variation in the interval between the first administration of these drugs and clinical response, and this may account for the inconsistencies in published trials. The type of drug and its dose may affect rate of response, as may biochemical factors, including acetylator and monoamine oxidase status. To obtain maximum benefit, a course of therapy with MAOIs should last for several months.

  19. In vivo low-level light therapy increases cytochrome oxidase in skeletal muscle.

    PubMed

    Hayworth, Christopher R; Rojas, Julio C; Padilla, Eimeira; Holmes, Genevieve M; Sheridan, Eva C; Gonzalez-Lima, F

    2010-01-01

    Low-level light therapy (LLLT) increases survival of cultured cells, improves behavioral recovery from neurodegeneration and speeds wound healing. These beneficial effects are thought to be mediated by upregulation of mitochondrial proteins, especially the respiratory enzyme cytochrome oxidase. However, the effects of in vivo LLLT on cytochrome oxidase in intact skeletal muscle have not been previously investigated. We used a sensitive method for enzyme histochemistry of cytochrome oxidase to examine the rat temporalis muscle 24 h after in vivo LLLT. The findings showed for the first time that in vivo LLLT induced a dose- and fiber type-dependent increase in cytochrome oxidase in muscle fibers. LLLT was particularly effective at enhancing the aerobic capacity of intermediate and red fibers. The findings suggest that LLLT may enhance the oxidative energy metabolic capacity of different types of muscle fibers, and that LLLT may be used to enhance the aerobic potential of skeletal muscle.

  20. Mammalian monoamine-oxidizing enzymes, with special reference to benzylamine oxidase in human tissues.

    PubMed

    Lewinsohn, R

    1984-01-01

    A review is presented of the monoamine-oxidizing enzymes with special reference to the activity of benzylamine oxidase (BzAO) in human tissues. Methods of study of amine oxidases, properties (chiefly of BzAO) and some problems concerning substrate and inhibitor specificity and multiple forms of monoamine oxidase (MAO) are surveyed. The substrate specificity of human plasma BzAO is compared with that of amine-oxidizing enzymes in plasma or serum of other species. Correlations of plasma BzAO and platelet MAO activity with clinical findings are discussed. The distribution of amine oxidase activities in solid human tissues is reviewed, in particular BzAO in blood vessels and richly-vascularized tissues, as well as kinetic constants and altered patterns of activity of BzAO in human atherosclerosis. Activities of the amine oxidases in non-vascular smooth muscle, in cultured cells, and in various tissues related to human gestation, are discussed. The present knowledge of BzAO is discussed in terms of its possible clinical relevance to several human disease states, and the importance of the enzyme in the human body.

  1. Herbivore-plant interactions: mixed-function oxidases and secondary plant substances.

    PubMed

    Brattsten, L B; Wilkinson, C F; Eisner, T

    1977-06-17

    The mixed-function oxidases of a polyphagous insect larva (the southern armyworm, Spodoptera eridania) were found to be induced by a diversity of secondary plant substances. The induction proceeds rapidly and in response to a small quantity of secondary substance. Following induction, the larva is less susceptible to dietary poisoning. It is argued that mixed-function oxidases play a major role in protecting herbivores against chemical stress from secondary plant substances.

  2. Snake Venom L-Amino Acid Oxidases: Trends in Pharmacology and Biochemistry

    PubMed Central

    Izidoro, Luiz Fernando M.; Sobrinho, Juliana C.; Mendes, Mirian M.; Costa, Tássia R.; Grabner, Amy N.; Rodrigues, Veridiana M.; da Silva, Saulo L.; Zanchi, Fernando B.; Zuliani, Juliana P.; Fernandes, Carla F. C.; Calderon, Leonardo A.; Stábeli, Rodrigo G.; Soares, Andreimar M.

    2014-01-01

    L-amino acid oxidases are enzymes found in several organisms, including venoms of snakes, where they contribute to the toxicity of ophidian envenomation. Their toxicity is primarily due to enzymatic activity, but other mechanisms have been proposed recently which require further investigation. L-amino acid oxidases exert biological and pharmacological effects, including actions on platelet aggregation and the induction of apoptosis, hemorrhage, and cytotoxicity. These proteins present a high biotechnological potential for the development of antimicrobial, antitumor, and antiprotozoan agents. This review provides an overview of the biochemical properties and pharmacological effects of snake venom L-amino acid oxidases, their structure/activity relationship, and supposed mechanisms of action described so far. PMID:24738050

  3. NADPH Oxidase-Driven Phagocyte Recruitment Controls Candida albicans Filamentous Growth and Prevents Mortality

    PubMed Central

    Brothers, Kimberly M.; Gratacap, Remi L.; Barker, Sarah E.; Newman, Zachary R.; Norum, Ashley; Wheeler, Robert T.

    2013-01-01

    Candida albicans is a human commensal and clinically important fungal pathogen that grows as both yeast and hyphal forms during human, mouse and zebrafish infection. Reactive oxygen species (ROS) produced by NADPH oxidases play diverse roles in immunity, including their long-appreciated function as microbicidal oxidants. Here we demonstrate a non-traditional mechanistic role of NADPH oxidase in promoting phagocyte chemotaxis and intracellular containment of fungi to limit filamentous growth. We exploit the transparent zebrafish model to show that failed NADPH oxidase-dependent phagocyte recruitment to C. albicans in the first four hours post-infection permits fungi to germinate extracellularly and kill the host. We combine chemical and genetic tools with high-resolution time-lapse microscopy to implicate both phagocyte oxidase and dual-specific oxidase in recruitment, suggesting that both myeloid and non-myeloid cells promote chemotaxis. We show that early non-invasive imaging provides a robust tool for prognosis, strongly connecting effective early immune response with survival. Finally, we demonstrate a new role of a key regulator of the yeast-to-hyphal switching program in phagocyte-mediated containment, suggesting that there are species-specific methods for modulation of NADPH oxidase-independent immune responses. These novel links between ROS-driven chemotaxis and fungal dimorphism expand our view of a key host defense mechanism and have important implications for pathogenesis. PMID:24098114

  4. A Novel Colletotrichum graminicola Raffinose Oxidase in the AA5 Family

    PubMed Central

    Mollerup, Filip; Parikka, Kirsti; Koutaniemi, Sanna; Boer, Harry; Juvonen, Minna; Master, Emma; Tenkanen, Maija; Kruus, Kristiina

    2017-01-01

    ABSTRACT We describe here the identification and characterization of a copper radical oxidase from auxiliary activities family 5 (AA5_2) that was distinguished by showing preferential activity toward raffinose. Despite the biotechnological potential of carbohydrate oxidases from family AA5, very few members have been characterized. The gene encoding raffinose oxidase from Colletotrichum graminicola (CgRaOx; EC 1.1.3.−) was identified utilizing a bioinformatics approach based on the known modular structure of a characterized AA5_2 galactose oxidase. CgRaOx was expressed in Pichia pastoris, and the purified enzyme displayed the highest activity on the trisaccharide raffinose, whereas the activity on the disaccharide melibiose was three times lower and more than ten times lower activity was detected on d-galactose at a 300 mM substrate concentration. Thus, the substrate preference of CgRaOx was distinguished clearly from the substrate preferences of the known galactose oxidases. The site of oxidation for raffinose was studied by 1H nuclear magnetic resonance and mass spectrometry, and we confirmed that the hydroxyl group at the C-6 position was oxidized to an aldehyde and that in addition uronic acid was produced as a side product. A new electrospray ionization mass spectrometry method for the identification of C-6 oxidized products was developed, and the formation mechanism of the uronic acid was studied. CgRaOx presented a novel activity pattern in the AA5 family. IMPORTANCE Currently, there are only a few characterized members of the CAZy AA5 protein family. These enzymes are interesting from an application point of view because of their ability to utilize the cheap and abundant oxidant O2 without the requirement of complex cofactors such as FAD or NAD(P). Here, we present the identification and characterization of a novel AA5 member from Colletotrichum graminicola. As discussed in the present study, the bioinformatics approach using the modular structure of

  5. Treating alcoholism as a chronic disease: approaches to long-term continuing care.

    PubMed

    McKay, James R; Hiller-Sturmhofel, Susanne

    2011-01-01

    For many patients, alcohol and other drug (AOD) use disorders are chronic, recurring conditions involving multiple cycles of treatment, abstinence, and relapse. To disrupt this cycle, treatment can include continuing care to reduce the risk of relapse. The most commonly used treatment approach is initial intensive inpatient or outpatient care based on 12-step principles, followed by continuing care involving self-help groups, 12-step group counseling, or individual therapy. Although these programs can be effective, many patients drop out of initial treatment or do not complete continuing care. Thus, researchers and clinicians have begun to develop alternative approaches to enhance treatment retention in both initial and continuing care. One focus of these efforts has been the design of extended treatment models. These approaches increasingly blur the distinction between initial and continuing care and aim to prolong treatment participation by providing a continuum of care. Other researchers have focused on developing alternative treatment strategies (e.g., telephone-based interventions) that go beyond traditional settings and adaptive treatment algorithms that may improve outcomes for clients who do not respond well to traditional approaches.

  6. Proposed structural basis of interaction of piperine and related compounds with monoamine oxidases.

    PubMed

    Rahman, Taufiq; Rahmatullah, Mohammed

    2010-01-15

    Several studies have revealed piperine and a few related compounds as potent inhibitors of monoamine oxidases without delineating the underlying mechanism. Using in silico modelling, we propose a structural basis of such activity by showing that these compounds can successfully dock into the inhibitor binding pockets of human monoamine oxidase isoforms with predicted affinities comparable to some known inhibitors. The results therefore suggest that piperine can be a promising lead for developing novel monoamine oxidase inhibitors. Copyright 2009 Elsevier Ltd. All rights reserved.

  7. Targeting NADPH oxidase decreases oxidative stress in the transgenic sickle cell mouse penis.

    PubMed

    Musicki, Biljana; Liu, Tongyun; Sezen, Sena F; Burnett, Arthur L

    2012-08-01

    Sickle cell disease (SCD) is a state of chronic vasculopathy characterized by endothelial dysfunction and increased oxidative stress, but the sources and mechanisms responsible for reactive oxygen species (ROS) production in the penis are unknown. We evaluated whether SCD activates NADPH oxidase, induces endothelial nitric oxide synthase (eNOS) uncoupling, and decreases antioxidants in the SCD mouse penis. We further tested the hypothesis that targeting NADPH oxidase decreases oxidative stress in the SCD mouse penis. SCD transgenic (sickle) mice were used as an animal model of SCD. Hemizygous (hemi) mice served as controls. Mice received an NADPH oxidase inhibitor apocynin (10 mM in drinking water) or vehicle. Penes were excised at baseline for molecular studies. Markers of oxidative stress (4-hydroxy-2-nonenal [HNE]), sources of ROS (eNOS uncoupling and NADPH oxidase subunits p67(phox) , p47(phox) , and gp91(phox) ), and enzymatic antioxidants (superoxide dismutase [SOD]1, SOD2, catalase, and glutathione peroxidase-1 [GPx1]) were measured by Western blot in penes. Sources of ROS, oxidative stress, and enzymatic antioxidants in the SCD penis. Relative to hemi mice, SCD increased (P<0.05) protein expression of NADPH oxidase subunits p67(phox) , p47(phox) , and gp91(phox) , 4-HNE-modified proteins, induced eNOS uncoupling, and reduced Gpx1 expression in the penis. Apocynin treatment of sickle mice reversed (P<0.05) the abnormalities in protein expressions of p47(phox) , gp91(phox) (but not p67(phox) ) and 4-HNE, but only slightly (P>0.05) prevented eNOS uncoupling in the penis. Apocynin treatment of hemi mice did not affect any of these parameters. NADPH oxidase and eNOS uncoupling are sources of oxidative stress in the SCD penis; decreased GPx1 further contributes to oxidative stress. Inhibition of NADPH oxidase upregulation decreases oxidative stress, implying a major role for NADPH oxidase as a ROS source and a potential target for improving vascular function in

  8. Targeting NADPH Oxidase Decreases Oxidative Stress in the Transgenic Sickle Cell Mouse Penis

    PubMed Central

    Musicki, Biljana; Liu, Tongyun; Sezen, Sena F.; Burnett, Arthur L.

    2012-01-01

    Introduction Sickle cell disease (SCD) is a state of chronic vasculopathy characterized by endothelial dysfunction and increased oxidative stress, but the sources and mechanisms responsible for reactive oxygen species (ROS) production in the penis are unknown. Aims We evaluated whether SCD activates NADPH oxidase, induces endothelial nitric oxide synthase (eNOS) uncoupling, and decreases antioxidants in the SCD mouse penis. We further tested the hypothesis that targeting NADPH oxidase decreases oxidative stress in the SCD mouse penis. Methods SCD transgenic (sickle) mice were used as an animal model of SCD. Hemizygous (hemi) mice served as controls. Mice received an NADPH oxidase inhibitor apocynin (10 mM in drinking water) or vehicle. Penes were excised at baseline for molecular studies. Markers of oxidative stress (4-hydroxy-2-nonenal [HNE]), sources of ROS (eNOS uncoupling and NADPH oxidase subunits p67phox, p47phox, and gp91phox), and enzymatic antioxidants (superoxide dismutase [SOD]1, SOD2, catalase, and glutathione peroxidase-1 [GPx1]) were measured by Western blot in penes. Main Outcome Measures Sources of ROS, oxidative stress, and enzymatic antioxidants in the SCD penis. Results Relative to hemi mice, SCD increased (P < 0.05) protein expression of NADPH oxidase subunits p67phox, p47phox, and gp91phox, 4-HNE-modified proteins, induced eNOS uncoupling, and reduced Gpx1 expression in the penis. Apocynin treatment of sickle mice reversed (P < 0.05) the abnormalities in protein expressions of p47phox, gp91phox (but not p67phox) and 4-HNE, but only slightly (P > 0.05) prevented eNOS uncoupling in the penis. Apocynin treatment of hemi mice did not affect any of these parameters. Conclusion NADPH oxidase and eNOS uncoupling are sources of oxidative stress in the SCD penis; decreased GPx1 further contributes to oxidative stress. Inhibition of NADPH oxidase upregulation decreases oxidative stress, implying a major role for NADPH oxidase as a ROS source and a

  9. Multi-Copper Oxidases and Human Iron Metabolism

    PubMed Central

    Vashchenko, Ganna; MacGillivray, Ross T. A.

    2013-01-01

    Multi-copper oxidases (MCOs) are a small group of enzymes that oxidize their substrate with the concomitant reduction of dioxygen to two water molecules. Generally, multi-copper oxidases are promiscuous with regards to their reducing substrates and are capable of performing various functions in different species. To date, three multi-copper oxidases have been detected in humans—ceruloplasmin, hephaestin and zyklopen. Each of these enzymes has a high specificity towards iron with the resulting ferroxidase activity being associated with ferroportin, the only known iron exporter protein in humans. Ferroportin exports iron as Fe2+, but transferrin, the major iron transporter protein of blood, can bind only Fe3+ effectively. Iron oxidation in enterocytes is mediated mainly by hephaestin thus allowing dietary iron to enter the bloodstream. Zyklopen is involved in iron efflux from placental trophoblasts during iron transfer from mother to fetus. Release of iron from the liver relies on ferroportin and the ferroxidase activity of ceruloplasmin which is found in blood in a soluble form. Ceruloplasmin, hephaestin and zyklopen show distinctive expression patterns and have unique mechanisms for regulating their expression. These features of human multi-copper ferroxidases can serve as a basis for the precise control of iron efflux in different tissues. In this manuscript, we review the biochemical and biological properties of the three human MCOs and discuss their potential roles in human iron homeostasis. PMID:23807651

  10. Function of the Pyruvate Oxidase-Lactate Oxidase Cascade in Interspecies Competition between Streptococcus oligofermentans and Streptococcus mutans

    PubMed Central

    Liu, Lei

    2012-01-01

    Complex interspecies interactions occur constantly between oral commensals and the opportunistic pathogen Streptococcus mutans in dental plaque. Previously, we showed that oral commensal Streptococcus oligofermentans possesses multiple enzymes for H2O2 production, especially lactate oxidase (Lox), allowing it to out-compete S. mutans. In this study, through extensive biochemical and genetic studies, we identified a pyruvate oxidase (pox) gene in S. oligofermentans. A pox deletion mutant completely lost Pox activity, while ectopically expressed pox restored activity. Pox was determined to produce most of the H2O2 in the earlier growth phase and log phase, while Lox mainly contributed to H2O2 production in stationary phase. Both pox and lox were expressed throughout the growth phase, while expression of the lox gene increased by about 2.5-fold when cells entered stationary phase. Since lactate accumulation occurred to a large degree in stationary phase, the differential Pox- and Lox-generated H2O2 can be attributed to differential gene expression and substrate availability. Interestingly, inactivation of pox causes a dramatic reduction in H2O2 production from lactate, suggesting a synergistic action of the two oxidases in converting lactate into H2O2. In an in vitro two-species biofilm experiment, the pox mutant of S. oligofermentans failed to inhibit S. mutans even though lox was active. In summary, S. oligofermentans develops a Pox-Lox synergy strategy to maximize its H2O2 formation so as to win the interspecies competition. PMID:22287002

  11. Alcoholism and Alcohol Abuse

    MedlinePlus

    ... their drinking causes distress and harm. It includes alcoholism and alcohol abuse. Alcoholism, or alcohol dependence, is a disease that causes ... the liver, brain, and other organs. Drinking during pregnancy can harm your baby. Alcohol also increases the ...

  12. Direct comparison of gluco-oligosaccharide oxidase variants and glucose oxidase: substrate range and H2O2 stability.

    PubMed

    Vuong, Thu V; Foumani, Maryam; MacCormick, Benjamin; Kwan, Rachel; Master, Emma R

    2016-11-21

    Glucose oxidase (GO) activity is generally restricted to glucose and is susceptible to inactivation by H 2 O 2 . By comparison, the Y300A variant of gluco-oligosaccharide oxidase (GOOX) from Sarocladium strictum showed broader substrate range and higher H 2 O 2 stability. Specifically, Y300A exhibited up to 40 times higher activity on all tested sugars except glucose, compared to GO. Moreover, fusion of the Y300A variant to a family 22 carbohydrate binding module from Clostridium thermocellum (CtCBM22A) nearly doubled its catalytic efficiency on glucose, while retaining significant activity on oligosaccharides. In the presence of 200 mM of H 2 O 2 , the recombinant CtCBM22A_Y300A retained 80% of activity on glucose and 100% of activity on cellobiose, the preferred substrate for this enzyme. By contrast, a commercial glucose oxidase reported to contain ≤0.1 units catalase/ mg protein, retained 60% activity on glucose under the same conditions. GOOX variants appear to undergo a different mechanism of inactivation, as a loss of histidine instead of methionine was observed after H 2 O 2 incubation. The addition of CtCBM22A also promoted functional binding of the fusion enzyme to xylan, facilitating its simultaneous purification and immobilization using edible oat spelt xylan, which might benefit the usage of this enzyme preparation in food and baking applications.

  13. Size-selective QD@MOF core-shell nanocomposites for the highly sensitive monitoring of oxidase activities.

    PubMed

    Wang, Ke; Li, Nan; Zhang, Jing; Zhang, Zhiqi; Dang, Fuquan

    2017-01-15

    In this work, we proposed a novel and facile method to monitor oxidase activities based on size-selective fluorescent quantum dot (QD)@metal-organic framework (MOF) core-shell nanocomposites (CSNCPs). The CSNCPs were synthesized from ZIF-8 and CdTe QDs in aqueous solution in 40min at room temperature with stirring. The prepared CdTe@ZIF-8 CSNCPs , which have excellent water dispersibility and stability, displays distinct fluorescence responses to hole scavengers of different molecular sizes (e.g., H 2 O 2 , substrate, and oxidase) due to the aperture limitation of the ZIF-8 shell. H 2 O 2 can efficiently quench the fluorescence of CdTe@ZIF-8 CSNCPs over a linearity range of 1-100nM with a detection limit of 0.29nM, whereas large molecules such as substrate and oxidase have very little effect on its fluorescence. Therefore, the highly sensitive detection of oxidase activities was achieved by monitoring the fluorescence quenching of CdTe@ZIF-8 CSNCPs by H 2 O 2 produced in the presence of substrate and oxidase, which is proportional to the oxidase activities. The linearity ranges of the uricase and glucose oxidase activity are 0.1-50U/L and 1-100U/L, respectively, and their detection limits are 0.024U/L and 0.26U/L, respectively. Therefore, the current QD@MOF CSNCPs based sensing system is a promising, widely applicable means of monitoring oxidase activities in biochemical research. Copyright © 2016 Elsevier B.V. All rights reserved.

  14. Steady-state kinetics of substrate binding and iron release in tomato ACC oxidase.

    PubMed

    Thrower, J S; Blalock, R; Klinman, J P

    2001-08-14

    1-Aminocyclopropane-1-carboxylate oxidase (ACC oxidase) catalyzes the last step in the biosynthetic pathway of the plant hormone, ethylene. This unusual reaction results in the oxidative ring cleavage of 1-aminocyclopropane carboxylate (ACC) into ethylene, cyanide, and CO2 and requires ferrous ion, ascorbate, and molecular oxygen for catalysis. A new purification procedure and assay method have been developed for tomato ACC oxidase that result in greatly increased enzymatic activity. This method allowed us to determine the rate of iron release from the enzyme and the effect of the activator, CO2, on this rate. Initial velocity studies support an ordered kinetic mechanism where ACC binds first followed by O2; ascorbate can bind after O2 or possibly before ACC. This kinetic mechanism differs from one recently proposed for the ACC oxidase from avocado.

  15. Neovascularization in an arterio-venous loop-containing tissue engineering chamber: role of NADPH oxidase

    PubMed Central

    Jiang, F; Zhang, G; Hashimoto, I; Kumar, B S; Bortolotto, S; Morrison, W A; Dusting, G J

    2008-01-01

    Using an in vivo arterio-venous loop-containing tissue-engineering chamber, we have created a variety of vascularized tissue blocks, including functional myocardium. The viability of the transplanted cells is limited by the rate of neovascularization in the chamber. A Nox2-containing nicotinamide adenine dinucleotide phosphate (NADPH) oxidase is thought to have a critical role in ischaemic angiogenesis. In this study we investigated whether NADPH oxidase is involved in the neovascularization process in the tissue-engineering chamber. New blood vessels originating from the venous and the arterial ends of the loop could be identified after 3 days, and the vessel density (by lectin staining) peaked after 7 days and was maintained for at least 14 days. This was accompanied by granulation tissue formation and concomitant increase in the mRNA level of Nox4 NADPH oxidase. Although the total level of Nox2 mRNA in the chamber tissue decreased from day 3 to day 7, immunohistochemistry identified a strong expression of Nox2 in the endothelial cells of the new vessels. In human microvascular endothelial cells, the NADPH oxidase inhibitor apocynin reduced NADPH oxidase activity and inhibited the angiogenic responses in vitro. Local treatment with the NADPH oxidase inhibitors apocynin or gp91ds-tat peptide significantly suppressed the vessel growth in the chamber. In conclusion, NADPH oxidase-dependent redox signalling is important for neovascularization in this novel tissue-engineering chamber in vivo, and boosting this signalling might be a new approach to extending vascularization and tissue growth. PMID:19012731

  16. Recent advances in Parkinson's disease therapy: use of monoamine oxidase inhibitors.

    PubMed

    Henchcliffe, Claire; Schumacher, H Christian; Burgut, F Tuna

    2005-11-01

    Monoamine oxidase inhibitors inhibit dopamine metabolism and are therefore effective in treating Parkinson's disease, a condition associated with progressive striatal dopamine deficiency secondary to degeneration of dopaminergic neurons in the substantia nigra. Selegiline is currently the most widely used monoamine oxidase-B inhibitor for Parkinson's disease, but has a low and variable bioavailability, and is metabolized to L-methamphetamine and L-amphetamine that carry a risk for potential neurotoxicity. There are two new approaches that circumvent these potential disadvantages. First, selegiline orally disintegrating tablets provide a novel delivery form of selegiline, avoiding first pass metabolism by rapid absorption through the oral mucosa, thus leading to significantly lower plasma concentrations of L-metamphetamine and L-amphetamine. Selegiline orally disintegrating tablets prove to be clinically effective and safe in patients with moderately advanced Parkinson's disease. Second, rasagiline is a new monoamine oxidase inhibitor, without known neurotoxic metabolites. In large clinical trials, rasagiline proves effective as monotherapy in early Parkinson's disease, as well as adjunctive therapy to levodopa in advanced disease. Clinical data suggest, in addition, a disease-modifying effect of rasagiline that may correlate with neuroprotective activity of monoamine oxidase-B inhibitors in animal models of Parkinson's disease.

  17. Insights into proton translocation in cbb3 oxidase from MD simulations.

    PubMed

    Carvalheda, Catarina A; Pisliakov, Andrei V

    2017-05-01

    Heme-copper oxidases are membrane protein complexes that catalyse the final step of the aerobic respiration, namely the reduction of oxygen to water. The energy released during catalysis is coupled to the active translocation of protons across the membrane, which contributes to the establishment of an electrochemical gradient that is used for ATP synthesis. The distinctive C-type (or cbb 3 ) cytochrome c oxidases, which are mostly present in proteobacteria, exhibit a number of unique structural and functional features, including high catalytic activity at low oxygen concentrations. At the moment, the functioning mechanism of C-type oxidases, in particular the proton transfer/pumping mechanism presumably via a single proton channel, is still poorly understood. In this work we used all-atom molecular dynamics simulations and continuum electrostatics calculations to obtain atomic-level insights into the hydration and dynamics of a cbb 3 oxidase. We provide the details of the water dynamics and proton transfer pathways for both the "chemical" and "pumped" protons, and show that formation of protonic connections is strongly affected by the protonation state of key residues, namely H243, E323 and H337. Copyright © 2017 Elsevier B.V. All rights reserved.

  18. Expression Studies of Gibberellin Oxidases in Developing Pumpkin Seeds1

    PubMed Central

    Frisse, Andrea; Pimenta, Maria João; Lange, Theo

    2003-01-01

    Two cDNA clones, 3-ox and 2-ox, have been isolated from developing pumpkin (Cucurbita maxima) embryos that show significant amino acid homology to gibberellin (GA) 3-oxidases and 2-oxidases, respectively. Recombinant fusion protein of clone 3-ox converted GA12-aldehyde, GA12, GA15, GA24, GA25, and GA9 to GA14-aldehyde, GA14, GA37, GA36, GA13, and GA4, respectively. Recombinant 2-ox protein oxidized GA9, GA4, and GA1 to GA51, GA34, and GA8, respectively. Previously cloned GA 7-oxidase revealed additional 3β-hydroxylation activity of GA12. Transcripts of this gene were identified in endosperm and embryo of the developing seed by quantitative reverse transcriptase-polymerase chain reaction and localized in protoderm, root apical meristem, and quiescent center by in situ hybridization. mRNA of the previously cloned GA 20-oxidase from pumpkin seeds was localized in endosperm and in tissues of protoderm, ground meristem, and cotyledons of the embryo. However, transcripts of the recently cloned GA 20-oxidase from pumpkin seedlings were found all over the embryo, and in tissues of the inner seed coat at the micropylar end. Previously cloned GA 2β,3β-hydroxylase mRNA molecules were specifically identified in endosperm tissue. Finally, mRNA molecules of the 3-ox and 2-ox genes were found in the embryo only. 3-ox transcripts were localized in tissues of cotyledons, protoderm, and inner cell layers of the root apical meristem, and 2-ox transcripts were found in all tissues of the embryo except the root tips. These results indicate tissue-specific GA-biosynthetic pathways operating within the developing seed. PMID:12644672

  19. Inheritance of polyphenol oxidase activity in wheat breeding lines derived from matings of low polyphenol oxidase parents

    USDA-ARS?s Scientific Manuscript database

    Polyphenol oxidase (PPO) in grain plays a major role in time-dependent discoloration of wheat (Triticum aestivum L.) products, especially fresh noodles. Breeding wheat cultivars with low or nil PPO activity can reduce the undesirable product darkening. The low PPO line PI 117635 was crossed to two...

  20. NADPH Oxidase-Dependent Signaling in Endothelial Cells: Role in Physiology and Pathophysiology

    PubMed Central

    Ushio-Fukai, Masuko; Malik, Asrar B.

    2009-01-01

    Abstract Reactive oxygen species (ROS) including superoxide (O2·−) and hydrogen peroxide (H2O2) are produced endogenously in response to cytokines, growth factors; G-protein coupled receptors, and shear stress in endothelial cells (ECs). ROS function as signaling molecules to mediate various biological responses such as gene expression, cell proliferation, migration, angiogenesis, apoptosis, and senescence in ECs. Signal transduction activated by ROS, “oxidant signaling,” has received intense investigation. Excess amount of ROS contribute to various pathophysiologies, including endothelial dysfunction, atherosclerosis, hypertension, diabetes, and acute respiratory distress syndrome (ARDS). The major source of ROS in EC is a NADPH oxidase. The prototype phagaocytic NADPH oxidase is composed of membrane-bound gp91phox and p22hox, as well as cytosolic subunits such as p47phox, p67phox and small GTPase Rac. In ECs, in addition to all the components of phagocytic NADPH oxidases, homologues of gp91phox (Nox2) including Nox1, Nox4, and Nox5 are expressed. The aim of this review is to provide an overview of the emerging area of ROS derived from NADPH oxidase and oxidant signaling in ECs linked to physiological and pathophysiological functions. Understanding these mechanisms may provide insight into the NADPH oxidase and oxidant signaling components as potential therapeutic targets. Antioxid. Redox Signal. 11, 791–810. PMID:18783313

  1. Observations and modelling of enhanced AOD at Spitsbergen - long-range transport of biomass burning emissions from Alaska fires in July 2015

    NASA Astrophysics Data System (ADS)

    Struzewska, Joanna; Kaminski, Jacek W.; Markowicz, Krzysztof; Lisok, Justyna

    2017-04-01

    In the scope of the IAREA project (http://www.igf.fuw.edu.pl/iAREA) several measurement campaigns were undertaken at Svalbard in order to assess the variability of absorbing aerosols in this region during spring and summertime 2014-2016. During the summer campaign of 2015 a significant increase of aerosol concentrations and AOD were observed at NyAlesund station from 9 to 15 of July. This episode was connected with the long range transport of smoke plum emitted from biomass burning at Alaska. A tropospheric chemistry model GEM-AQ (Global Environmental Multiscale Air Quality) was used to analyse the pathways and vertical distribution of the biomass burning aerosol observed over Svalbard. The core of the model is based on a weather prediction model with environmental processes (chemistry and aerosols) implanted on-line and are interactive (i.e. providing feedback of chemistry on radiation and dynamics). Aerosol module includes 5 size-resolved types of aerosols: sulphates, black carbon, organic carbon, sea salt and mineral dust. Aerosol mass is distributed into 12 logarithmically spaced bins. Anthropogenic emissions developed by NILU in the ECLIPSE project were used for this study. Biomass burning emissions were applied using daily GFAS fields obtained from the Copernicus Services. We will present the GEM-AQ model results and comparison with available measurement. The GEM-AQ model correctly reproduced the inflow of polluted air masses and sharp increase of AOD values. PM10 analysis showed elevated concentration values up to 700 hPa level, with the maximum reaching 35 ppb at 900 hPa. Model analysis showed over Svalbard shows transport of a thick layer with enhanced extinction above the planetary boundary layer.

  2. Effect of voluntary alcohol consumption on Maoa expression in the mesocorticolimbic brain of adult male rats previously exposed to prolonged maternal separation.

    PubMed

    Bendre, M; Comasco, E; Nylander, I; Nilsson, K W

    2015-12-08

    Discordant associations between monoamine oxidase A (MAOA) genotype and high alcohol drinking have been reported in human and non-human primates. Environmental influences likely moderate genetic susceptibility. The biological basis for this interplay remains elusive, and inconsistencies call for translational studies in which conditions can be controlled and brain tissue is accessible. The present study investigated whether early life stress and subsequent adult episodic alcohol consumption affect Maoa expression in stress- and reward-related brain regions in the rat. Outbred Wistar rats were exposed to rearing conditions associated with stress (prolonged maternal separation) or no stress during early life, and given free choice between alcohol and/or water in adulthood. Transcript levels of Maoa were assessed in the ventral tegmental area, nucleus accumbens (NAc), medial prefrontal cortex, cingulate cortex, amygdala and dorsal striatum (DS). Blood was collected to assess corticosterone levels. After alcohol consumption, lower blood corticosterone and Maoa expression in the NAc and DS were found in rats exposed to early life stress compared with control rats. An interaction between early life stress and voluntary alcohol intake was found in the NAc. Alcohol intake before death correlated negatively with Maoa expression in DS in high alcohol-drinking rats exposed to early life stress. Maoa expression is sensitive to adulthood voluntary alcohol consumption in the presence of early life stress in outbred rats. These findings add knowledge of the molecular basis of the previously reported associations between early life stress, MAOA and susceptibility to alcohol misuse.

  3. Increased xanthine oxidase during labour--implications for oxidative stress.

    PubMed

    Many, A; Roberts, J M

    1997-11-01

    Xanthine dehydrogenase/oxidase (XDH/XO) produces uric acid. When in the oxidase form, this production is coupled with the generation of free radicals. Hypoxia-reperfusion enhances conversion of XDH to XO. Since the placenta is exposed to short periods of hypoxia reperfusion during labour, 17 placentae of pregnancy terminated by elective caesarean section and five placentae of pregnancies terminated by caesarean section during labour were examined for XDH/XO activity. It was found that XO activity was higher in the placentae of labouring women (P = 0.003), which suggests that labour enhances conversion of XDH to XO, facilitating free radical production.

  4. A Conserved Steroid Binding Site in Cytochrome c Oxidase

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

    Qin, Ling; Mills, Denise A.; Buhrow, Leann

    2010-09-02

    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.

  5. Production of a new D-amino acid oxidase from the fungus Fusarium oxysporum.

    PubMed

    Gabler, M; Fischer, L

    1999-08-01

    The fungus Fusarium oxysporum produced a D-amino acid oxidase (EC 1. 4.3.3) in a medium containing glucose as the carbon and energy source and ammonium sulfate as the nitrogen source. The specific D-amino acid oxidase activity was increased up to 12.5-fold with various D-amino acids or their corresponding derivatives as inducers. The best inducers were D-alanine (2.7 microkat/g of dry biomass) and D-3-aminobutyric acid (2.6 microkat/g of dry biomass). The addition of zinc ions was necessary to permit the induction of peroxisomal D-amino acid oxidase. Bioreactor cultivations were performed on a 50-liter scale, yielding a volumetric D-amino acid oxidase activity of 17 microkat liter(-1) with D-alanine as an inducer. Under oxygen limitation, the volumetric activity was increased threefold to 54 microkat liter(-1) (3,240 U liter(-1)).

  6. Reconstituted high-density lipoprotein suppresses leukocyte NADPH oxidase activation by disrupting lipid rafts.

    PubMed

    Peshavariya, Hitesh; Dusting, Gregory J; Di Bartolo, Belinda; Rye, Kerry-Anne; Barter, Philip J; Jiang, Fan

    2009-08-01

    Reconstituted discoidal high-density lipoprotein (rHDL) has potent vascular protective actions. Native HDL suppresses cellular generation of reactive oxygen species, whereas this antioxidant effect of rHDL is less clear. This study examined the effects of rHDL on NADPH oxidase, a major source of cellular superoxide generation, in both leukocytes and human umbilical vein endothelial cells. Superoxide was measured with lucigenin-enhanced chemiluminescence. Expression of NADPH oxidase sub-units was determined by real-time PCR. Pre-treatment of HL-60 cells with rHDL (10 and 25 microM) for 1 h significantly reduced phorbol 12-myristate 13-acetate-stimulated superoxide production. Treatment with rHDL for up to 24 h did not change the mRNA expression of NADPH oxidase sub-units. In HL-60 cells, depletion of cholesterol from the plasma membrane by methyl-beta-cyclodextrin mimicked the effect of rHDL, whereas cholesterol repletion blunted the effects of rHDL. Treatment with rHDL induced disruption of the lipid raft structures and blunted PMA-induced redistribution of p47phox into lipid rafts. In contrast, treatment of endothelial cells with rHDL for up to 18 h had no effect on either basal or tumour necrosis factor-alpha-stimulated NADPH oxidase activity, but markedly suppressed the cytokine-induced expression of proinflammatory adhesion molecules. The results suggest that rHDL inhibits NADPH oxidase activation in leukocytes, probably by interrupting the assembly of NADPH oxidase sub-units at the lipid rafts. This effect may contribute to the vascular protective actions of rHDL against inflammation-mediated oxidative damage.

  7. An updated patent review: xanthine oxidase inhibitors for the treatment of hyperuricemia and gout (2011-2015).

    PubMed

    Ojha, Ritu; Singh, Jagjeet; Ojha, Anu; Singh, Harbinder; Sharma, Sahil; Nepali, Kunal

    2017-03-01

    Xanthine oxidase (XO) is a versatile molybdoflavoprotein, widely distributed, occurring in milk, kidney, lung, heart, and vascular endothelium. Catalysis by XO to produce uric acid and reactive oxygen species leads to many diseases. Anti hyperuricemic therapy by xanthine oxidase inhibitors has been mainly employed for the treatment of gout. Area covered: This review covers the patent literature (2011-2015) and also presents the interesting strategies/rational approaches employed for the design of xanthine oxidase inhibitors reported recently. Expert opinion: Recent literature indicates that various non purine scaffolds have been extensively investigated for xanthine oxidase inhibition. The significant potential endowed by heteroaryl based compounds, in particularly fused heterocycles clearly highlights their clinical promise and the need for detailed investigation. Studies by various research groups have also revealed that the flavone framework is open for isosteric replacements and structural modifications for yielding potent non purine xanthine oxidase inhibitors. In addition, various plant extracts recently reported to possess significant xanthine oxidase inhibitory potential presents enough promise to initiate a screening program for the identification of other plant extracts and phytoconstituents possessing inhibitory potential towards the enzyme.

  8. NADPH Oxidase versus Mitochondria-Derived ROS in Glucose-Induced Apoptosis of Pericytes in Early Diabetic Retinopathy

    PubMed Central

    Mustapha, Nik M.; Tarr, Joanna M.; Kohner, Eva M.; Chibber, Rakesh

    2010-01-01

    Objectives. Using apocynin (inhibitor of NADPH oxidase), and Mitoquinol 10 nitrate (MitoQ; mitochondrial-targeted antioxidant), we addressed the importance of mitochondria versus NADPH oxidase-derived ROS in glucose-induced apoptosis of pericytes. Methods. NADPH oxidase was localised using Western blot analysis and cytochrome C reduction assay. Apoptosis was detected by measuring caspase-3 activity. Intracellular glucose concentration, ROS formation and Nε-(carboxymethyl) lysine (CML) content were measured using Amplex Red assay kit, dihydroethidium (DHE), and competitive immunoabsorbant enzyme-linked assay (ELISA), respectively. Results. NADPH oxidase was localised in the cytoplasm of pericytes suggesting ROS production within intracellular compartments. High glucose (25 mM) significantly increased apoptosis, intracellular glucose concentration, and CML content. Apoptosis was associated with increased gp91phox expression, activity of NADPH oxidase, and intracellular ROS production. Apocynin and not MitoQ significantly blunted the generation of ROS, formation of intracellular CML and apoptosis. Conclusions. NADPH oxidase and not mitochondria-derived ROS is responsible for the accelerated apoptosis of pericytes in diabetic retinopathy. PMID:20652059

  9. Genetics Home Reference: isolated sulfite oxidase deficiency

    MedlinePlus

    ... Metabolic Disorders (CLIMB) March of Dimes: Amino Acid Metabolism Disorders The Compassionate Friends GeneReviews (1 link) Isolated Sulfite Oxidase Deficiency ClinicalTrials.gov (1 link) ClinicalTrials.gov Scientific Articles on PubMed (1 link) PubMed OMIM (1 link) ...

  10. Purification, characterization and amino acid content of cholesterol oxidase produced by Streptomyces aegyptia NEAE 102.

    PubMed

    El-Naggar, Noura El-Ahmady; Deraz, Sahar F; Soliman, Hoda M; El-Deeb, Nehal M; El-Shweihy, Nancy M

    2017-03-29

    There is an increasing demand on cholesterol oxidase for its various industrial and clinical applications. The current research was focused on extracellular cholesterol oxidase production under submerged fermentation by a local isolate previously identified as Streptomyces aegyptia NEAE 102. The crude enzyme extract was purified by two purification steps, protein precipitation using ammonium sulfate followed by ion exchange chromatography using DEAE Sepharose CL-6B. The kinetic parameters of purified cholesterol oxidase from Streptomyces aegyptia NEAE 102 were studied. The best conditions for maximum cholesterol oxidase activity were found to be 105 min of incubation time, an initial pH of 7 and temperature of 37 °C. The optimum substrate concentration was found to be 0.4 mM. The higher thermal stability behavior of cholesterol oxidase was at 50 °C. Around 63.86% of the initial activity was retained by the enzyme after 20 min of incubation at 50 °C. The apparent molecular weight of the purified enzyme as sized by sodium dodecyl sulphate-polyacryalamide gel electrophoresis was approximately 46 KDa. On DEAE Sepharose CL-6B column cholesterol oxidase was purified to homogeneity with final specific activity of 16.08 U/mg protein and 3.14-fold enhancement. The amino acid analysis of the purified enzyme produced by Streptomyces aegyptia NEAE 102 illustrated that, cholesterol oxidase is composed of 361 residues with glutamic acid as the most represented amino acid with concentration of 11.49 μg/mL. Taking into account the extracellular production, wide pH tolerance, thermal stability and shelf life, cholesterol oxidase produced by Streptomyces aegyptia NEAE 102 suggested that the enzyme could be industrially useful.

  11. PKC delta and NADPH oxidase in retinoic acid-induced neuroblastoma cell differentiation.

    PubMed

    Nitti, Mariapaola; Furfaro, Anna Lisa; Cevasco, Claudia; Traverso, Nicola; Marinari, Umberto Maria; Pronzato, Maria Adelaide; Domenicotti, Cinzia

    2010-05-01

    The role of reactive oxygen species (ROS) in the regulation of signal transduction processes has been well established in many cell types and recently the fine tuning of redox signalling in neurons received increasing attention. With regard to this, the involvement of NADPH oxidase (NOX) in neuronal pathophysiology has been proposed but deserves more investigation. In the present study, we used SH-SY5Y neuroblastoma cells to analyse the role of NADPH oxidase in retinoic acid (RA)-induced differentiation, pointing out the involvement of protein kinase C (PKC) delta in the activation of NOX. Retinoic acid induces neuronal differentiation as revealed by the increased expression of MAP2, the decreased cell doubling rate, and the gain in neuronal morphological features and these events are accompanied by the increased expression level of PKC delta and p67(phox), one of the components of NADPH oxidase. Using DPI to inhibit NOX activity we show that retinoic acid acts through this enzyme to induce morphological changes linked to the differentiation. Moreover, using rottlerin to inhibit PKC delta or transfection experiments to overexpress it, we show that retinoic acid acts through this enzyme to induce MAP2 expression and to increase p67(phox) membrane translocation leading to NADPH oxidase activation. These findings identify the activation of PKC delta and NADPH oxidase as crucial steps in RA-induced neuroblastoma cell differentiation. 2010 Elsevier Inc. All rights reserved.

  12. Structural insights into electron transfer in caa3-type cytochrome oxidase

    PubMed Central

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

    2012-01-01

    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 c2–4. Here we report the crystal structure of the caa3-type cytochrome oxidase from Thermus thermophilus, which has a covalently tethered cytochrome c domain. Crystals were grown in a bicontinuous mesophase using a synthetic short-chain monoacylglycerol as the hosting lipid. From the electron density map, at 2.36 Å resolution, a novel integral membrane subunit and a native glycoglycerophospholipid embedded in the complex were identified. Contrary to previous electron transfer mechanisms observed for soluble cytochrome c, the structure reveals the architecture of the electron transfer complex for the fused cupredoxin/cytochrome c domain which implicates different sites on cytochrome c for electron entry and exit. Support for an alternative to the classical proton gate characteristic of this HCO class is presented. PMID:22763450

  13. Molecular characterization of the fatty alcohol oxidation pathway for wax-ester mobilization in germinated jojoba seeds.

    PubMed

    Rajangam, Alex S; Gidda, Satinder K; Craddock, Christian; Mullen, Robert T; Dyer, John M; Eastmond, Peter J

    2013-01-01

    Jojoba (Simmondsia chinensis) is the only plant species known to use liquid wax esters (WEs) as a primary seed storage reserve. Upon germination, WE hydrolysis releases very-long-chain fatty alcohols, which must be oxidized to fatty acids by the sequential action of a fatty alcohol oxidase (FAO) and a fatty aldehyde dehydrogenase (FADH) before they can be β-oxidized. Here, we describe the cloning and characterization of genes for each of these two activities. Jojoba FAO and FADH are 52% and 68% identical to Arabidopsis (Arabidopsis thaliana) FAO3 and ALDH3H1, respectively. The genes are expressed most strongly in the cotyledons of jojoba seedlings following germination, but transcripts can also be detected in vegetative tissues. Proteomic analysis indicated that the FAO and FADH proteins can be detected on wax bodies, but they localized to the endoplasmic reticulum when they were expressed as amino-terminal green fluorescent protein fusions in tobacco (Nicotiana tabacum) leaves. Recombinant jojoba FAO and FADH proteins are active on very-long-chain fatty alcohol and fatty aldehyde substrates, respectively, and have biochemical properties consistent with those previously reported in jojoba cotyledons. Coexpression of jojoba FAO and FADH in Arabidopsis enhanced the in vivo rate of fatty alcohol oxidation more than 4-fold. Taken together, our data suggest that jojoba FAO and FADH constitute the very-long-chain fatty alcohol oxidation pathway that is likely to be necessary for efficient WE mobilization following seed germination.

  14. Crystal Structures of Intermediates in the Nitroalkane Oxidase Reaction

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

    Heroux, A.; Bozinovski, D; Valley, M

    2009-01-01

    The flavoenzyme nitroalkane oxidase is a member of the acyl-CoA dehydrogenase superfamily. Nitroalkane oxidase catalyzes the oxidation of neutral nitroalkanes to nitrite and the corresponding aldehydes or ketones. Crystal structures to 2.2 {angstrom} resolution or better of enzyme complexes with bound substrates and of a trapped substrate-flavin adduct are described. The D402N enzyme has no detectable activity with neutral nitroalkanes. The structure of the D402N enzyme crystallized in the presence of 1-nitrohexane or 1-nitrooctane shows the presence of the substrate in the binding site. The aliphatic chain of the substrate extends into a tunnel leading to the enzyme surface. Themore » oxygens of the substrate nitro group interact both with amino acid residues and with the 2'-hydroxyl of the FAD. When nitroalkane oxidase oxidizes nitroalkanes in the presence of cyanide, an electrophilic flavin imine intermediate can be trapped (Valley, M. P., Tichy, S. E., and Fitzpatrick, P. F. (2005) J. Am. Chem. Soc. 127, 2062-2066). The structure of the enzyme trapped with cyanide during oxidation of 1-nitrohexane shows the presence of the modified flavin. A continuous hydrogen bond network connects the nitrogen of the CN-hexyl-FAD through the FAD 2'-hydroxyl to a chain of water molecules extending to the protein surface. Together, our complementary approaches provide strong evidence that the flavin cofactor is in the appropriate oxidation state and correlates well with the putative intermediate state observed within each of the crystal structures. Consequently, these results provide important structural descriptions of several steps along the nitroalkane oxidase reaction cycle.« less

  15. A gene encoding the plant-like alternative oxidase is present in Phytomonas but absent in Leishmania spp.

    PubMed

    Van Hellemond, J J; Simons, B; Millenaar, F F; Tielens, A G

    1998-01-01

    The constituents of the respiratory chain are believed to differ among the trypanosomatids; bloodstream stages of African trypanosomes and Phytomonas promastigotes oxidize ubiquinol by a ubiquinol:oxygen oxidoreductase, also known as alternative oxidase, whereas Leishmania spp. oxidize ubiquinol via a classic cytochrome-containing respiratory chain. The molecular basis for this elementary difference in ubiquinol oxidation by the mitochondrial electron-transport chain in distinct trypanosomatids was investigated. The presence of a gene encoding the plant-like alternative oxidase could be demonstrated in Phytomonas and Trypanosoma brucei, trypanosomatids that are known to contain alternative oxidase activity. Our results further demonstrated that Leishmania spp. lack a gene encoding the plant-like alternative oxidase, and therefore, all stages of Leishmania spp. will lack the alternative oxidase protein. The observed fundamental differences between the respiratory chains of distinct members of the trypanosomatid family are thus caused by the presence or absence of a gene encoding the plant-like alternative oxidase.

  16. Androgen receptor and monoamine oxidase polymorphism in wild bonobos

    PubMed Central

    Garai, Cintia; Furuichi, Takeshi; Kawamoto, Yoshi; Ryu, Heungjin; Inoue-Murayama, Miho

    2014-01-01

    Androgen receptor gene (AR), monoamine oxidase A gene (MAOA) and monoamine oxidase B gene (MAOB) have been found to have associations with behavioral traits, such as aggressiveness, and disorders in humans. However, the extent to which similar genetic effects might influence the behavior of wild apes is unclear. We examined the loci AR glutamine repeat (ARQ), AR glycine repeat (ARG), MAOA intron 2 dinucleotide repeat (MAin2) and MAOB intron 2 dinucleotide repeat (MBin2) in 32 wild bonobos, Pan paniscus, and compared them with those of chimpanzees, Pan troglodytes, and humans. We found that bonobos were polymorphic on the four loci examined. Both loci MAin2 and MBin2 in bonobos showed a higher diversity than in chimpanzees. Because monoamine oxidase influences aggressiveness, the differences between the polymorphisms of MAin2 and MBin2 in bonobos and chimpanzees may be associated with the differences in aggression between the two species. In order to understand the evolution of these loci and AR, MAOA and MAOB in humans and non-human primates, it would be useful to conduct future studies focusing on the potential association between aggressiveness, and other personality traits, and polymorphisms documented in bonobos. PMID:25606465

  17. Ultrafine carbon particles promote rotenone-induced dopamine neuronal loss through activating microglial NADPH oxidase

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

    Wang, Yinxi; Liu, Dan; Zhang, Huifeng

    Background: Atmospheric ultrafine particles (UFPs) and pesticide rotenone were considered as potential environmental risk factors for Parkinson's disease (PD). However, whether and how UFPs alone and in combination with rotenone affect the pathogenesis of PD remains largely unknown. Methods: Ultrafine carbon black (ufCB, a surrogate of UFPs) and rotenone were used individually or in combination to determine their roles in chronic dopaminergic (DA) loss in neuron-glia, and neuron-enriched, mix-glia cultures. Immunochemistry using antibody against tyrosine hydroxylase was performed to detect DA neuronal loss. Measurement of extracellular superoxide and intracellular reactive oxygen species (ROS) were performed to examine activation of NADPHmore » oxidase. Genetic deletion and pharmacological inhibition of NADPH oxidase and MAC-1 receptor in microglia were employed to examine their role in DA neuronal loss triggered by ufCB and rotenone. Results: In rodent midbrain neuron-glia cultures, ufCB and rotenone alone caused neuronal death in a dose-dependent manner. In particularly, ufCB at doses of 50 and 100 μg/cm{sup 2} induced significant loss of DA neurons. More importantly, nontoxic doses of ufCB (10 μg/cm{sup 2}) and rotenone (2 nM) induced synergistic toxicity to DA neurons. Microglial activation was essential in this process. Furthermore, superoxide production from microglial NADPH oxidase was critical in ufCB/rotenone-induced neurotoxicity. Studies in mix-glia cultures showed that ufCB treatment activated microglial NADPH oxidase to induce superoxide production. Firstly, ufCB enhanced the expression of NADPH oxidase subunits (gp91{sup phox}, p47{sup phox} and p40{sup phox}); secondly, ufCB was recognized by microglial surface MAC-1 receptor and consequently promoted rotenone-induced p47{sup phox} and p67{sup phox} translocation assembling active NADPH oxidase. Conclusion: ufCB and rotenone worked in synergy to activate NADPH oxidase in microglia, leading to

  18. Analysis of cellulase and polyphenol oxidase production by southern pine beetle associated fungi

    Treesearch

    Abduvali Valiev; Zumrut B. Ogel; Dier D. Klepzig

    2009-01-01

    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....

  19. Inhibition of the NADPH oxidase regulates HO-1 expression in chronic myeloid leukemia

    PubMed Central

    Singh, Melissa M.; Irwin, Mary E.; Gao, Yin; Ban, Kechen; Shi, Ping; Arlinghaus, Ralph B.; Amin, Hesham M.; Chandra, Joya

    2011-01-01

    Background Patients with blast crisis phase chronic myelogeneous leukemia (CML) have poor response to tyrosine kinase inhibitors designed to inhibit the BCR-ABL1 oncogene. Recent work has shown that heme oxygenase 1 (HO-1) expression is increased in BCR-ABL1 expressing cells and that inhibition of HO-1 in CML leads to reduced cellular growth suggesting HO-1 may be a plausible target for therapy. Here we sought to clarify the mechanism of HO-1 overexpression and the role of the NADPH oxidase as a contributor to this mechanism in CML. Methods HO-1 expression was evaluated in CML bone marrow specimens from patients in various stages of disease, in a transplant based model for CML and in CML cell lines. Chemical and genetic inhibition of the NADPH oxidase was carried out in CML cells. Results Blast crisis CML patient specimens displayed higher levels of HO-1 staining than chronic or accelerated phase. HO-1 upregulation in BCR-ABL1 expressing cells was suppressed by diphenyliodonium (DPI), a chemical inhibitor of the NADPH oxidase. Targeting the NADPH oxidase through RNAi to Rac1, a dominant negative Rac1 construct or an inhibitor of Rac1 activity also blunted HO-1 protein expression. Moreover, inhibition of the NADPH oxidase by RNAi directed towards p47phox similarly abrogated HO-1 levels. Conclusion BCR-ABL1 expression upregulates HO-1, a survival factor for CML cells. This upregulation is more pronounced in blast crisis CML relative to early stage disease and is mediated by the NADPH oxidase components Rac1 and p47phox. Expression of p47phox is increased in BCR-ABL1 expressing cells. PMID:22139798

  20. Functional Assembly of Soluble and Membrane Recombinant Proteins of Mammalian NADPH Oxidase Complex.

    PubMed

    Souabni, Hajer; Ezzine, Aymen; Bizouarn, Tania; Baciou, Laura

    2017-01-01

    Activation of phagocyte cells from an innate immune system is associated with a massive consumption of molecular oxygen to generate highly reactive oxygen species (ROS) as microbial weapons. This is achieved by a multiprotein complex, the so-called NADPH oxidase. The activity of phagocyte NADPH oxidase relies on an assembly of more than five proteins, among them the membrane heterodimer named flavocytochrome b 558 (Cytb 558 ), constituted by the tight association of the gp91 phox (also named Nox2) and p22 phox proteins. The Cytb 558 is the membrane catalytic core of the NADPH oxidase complex, through which the reducing equivalent provided by NADPH is transferred via the associated prosthetic groups (one flavin and two hemes) to reduce dioxygen into superoxide anion. The other major proteins (p47 phox , p67 phox , p40 phox , Rac) requisite for the complex activity are cytosolic proteins. Thus, the NADPH oxidase functioning relies on a synergic multi-partner assembly that in vivo can be hardly studied at the molecular level due to the cell complexity. Thus, a cell-free assay method has been developed to study the NADPH oxidase activity that allows measuring and eventually quantifying the ROS generation based on optical techniques following reduction of cytochrome c. This setup is a valuable tool for the identification of protein interactions, of crucial components and additives for a functional enzyme. Recently, this method was improved by the engineering and the production of a complete recombinant NADPH oxidase complex using the combination of purified proteins expressed in bacterial and yeast host cells. The reconstitution into artificial membrane leads to a fully controllable system that permits fine functional studies.

  1. NADPH oxidases in the arbuscular mycorrhizal symbiosis.

    PubMed

    Belmondo, Simone; Calcagno, Cristina; Genre, Andrea; Puppo, Alain; Pauly, Nicolas; Lanfranco, Luisa

    2016-01-01

    Plant NADPH oxidases are the major source of reactive oxygen species (ROS) that plays key roles as both signal and stressor in several plant processes, including defense responses against pathogens. ROS accumulation in root cells during arbuscular mycorrhiza (AM) development has raised the interest in understanding how ROS-mediated defense programs are modulated during the establishment of this mutualistic interaction. We have recently analyzed the expression pattern of 5 NADPH oxidase (also called RBOH) encoding genes in Medicago truncatula, showing that only one of them (MtRbohE) is specifically upregulated in arbuscule-containing cells. In line with this result, RNAi silencing of MtRbohE generated a strong alteration in root colonization, with a significant reduction in the number of arbusculated cells. On this basis, we propose that MtRBOHE-mediated ROS production plays a crucial role in the intracellular accommodation of arbuscules.

  2. NADPH oxidases in the arbuscular mycorrhizal symbiosis

    PubMed Central

    Belmondo, Simone; Calcagno, Cristina; Genre, Andrea; Puppo, Alain; Pauly, Nicolas; Lanfranco, Luisa

    2016-01-01

    ABSTRACT Plant NADPH oxidases are the major source of reactive oxygen species (ROS) that plays key roles as both signal and stressor in several plant processes, including defense responses against pathogens. ROS accumulation in root cells during arbuscular mycorrhiza (AM) development has raised the interest in understanding how ROS-mediated defense programs are modulated during the establishment of this mutualistic interaction. We have recently analyzed the expression pattern of 5 NADPH oxidase (also called RBOH) encoding genes in Medicago truncatula, showing that only one of them (MtRbohE) is specifically upregulated in arbuscule-containing cells. In line with this result, RNAi silencing of MtRbohE generated a strong alteration in root colonization, with a significant reduction in the number of arbusculated cells. On this basis, we propose that MtRBOHE-mediated ROS production plays a crucial role in the intracellular accommodation of arbuscules. PMID:27018627

  3. Xanthine oxidase biosensor for monitoring meat spoilage

    NASA Astrophysics Data System (ADS)

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

    2014-05-01

    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.

  4. Oxoferryl-porphyrin radical catalytic intermediate in cytochrome bd oxidases protects cells from formation of reactive oxygen species.

    PubMed

    Paulus, Angela; Rossius, Sebastiaan Gijsbertus Hendrik; Dijk, Madelon; de Vries, Simon

    2012-03-16

    The quinol-linked cytochrome bd oxidases are terminal oxidases in respiration. These oxidases harbor a low spin heme b(558) that donates electrons to a binuclear heme b(595)/heme d center. The reaction with O(2) and subsequent catalytic steps of the Escherichia coli cytochrome bd-I oxidase were investigated by means of ultra-fast freeze-quench trapping followed by EPR and UV-visible spectroscopy. After the initial binding of O(2), the O-O bond is heterolytically cleaved to yield a kinetically competent heme d oxoferryl porphyrin π-cation radical intermediate (compound I) magnetically interacting with heme b(595). Compound I accumulates to 0.75-0.85 per enzyme in agreement with its much higher rate of formation (~20,000 s(-1)) compared with its rate of decay (~1,900 s(-1)). Compound I is next converted to a short lived heme d oxoferryl intermediate (compound II) in a phase kinetically matched to the oxidation of heme b(558) before completion of the reaction. The results indicate that cytochrome bd oxidases like the heme-copper oxidases break the O-O bond in a single four-electron transfer without a peroxide intermediate. However, in cytochrome bd oxidases, the fourth electron is donated by the porphyrin moiety rather than by a nearby amino acid. The production of reactive oxygen species by the cytochrome bd oxidase was below the detection level of 1 per 1000 turnovers. We propose that the two classes of terminal oxidases have mechanistically converged to enzymes in which the O-O bond is broken in a single four-electron transfer reaction to safeguard the cell from the formation of reactive oxygen species.

  5. Immunological comparison of sulfite oxidase

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

    Pollock, V.; Barber, M.J.

    1991-03-11

    Polyclonal antibodies (rabbit), elicited against FPLC-purified chicken and rat liver sulfite oxidase (SO), have been examined for inhibition and binding to purified chicken (C), rat (R), bovine (B), alligator (A) and shark (S) liver enzymes. Anti-CSO IgG cross-reacted with all five enzymes, with varying affinities, in the order CSO=ASO{gt}RSO{gt}BSO{gt}SSO. Anti-ROS IgG also cross-reacted with all five enzymes in the order RSO{gt}CSO=ASO{gt}BSO{gt}SSO. Anti-CSO IgG inhibited sulfite:cyt. c reductase (S:CR), sulfite:ferricyanide reductase (S:FR) and sulfite:dichlorophenolindophenol reductase (S:DR) activities of CSO to different extents (S:CR{gt}S:FR=S:DR). Similar differential inhibition was found for anti-ROS IgG and RSO S:CR, S:FR and S:DR activities. Anti-CSO IgG inhibitedmore » S:CR activities in the order CSO=ASO{much gt}SSO{gt}BSO. RSO was uninhibited. For anti-RSO IgG the inhibition order was RSO{gt}SSO{gt}BSO{gt}ASO. CSO was uninhibited. Anti-CSO and RSO IgGs partially inhibited Chlorella nitrate reductase (NR). Minor cross-reactivity was found for xanthine oxidase. Common antigenic determinants for all five SO's and NR are indicated.« less

  6. Glutamate Excitotoxicity Linked to Spermine Oxidase Overexpression.

    PubMed

    Pietropaoli, Stefano; Leonetti, Alessia; Cervetto, Chiara; Venturini, Arianna; Mastrantonio, Roberta; Baroli, Giulia; Persichini, Tiziana; Colasanti, Marco; Maura, Guido; Marcoli, Manuela; Mariottini, Paolo; Cervelli, Manuela

    2018-02-03

    Excitotoxic stress has been associated with several different neurological disorders, and it is one of the main causes of neuronal degeneration and death. To identify new potential proteins that could represent key factors in excitotoxic stress and to study the relationship between polyamine catabolism and excitotoxic damage, a novel transgenic mouse line overexpressing spermine oxidase enzyme in the neocortex (Dach-SMOX) has been engineered. These transgenic mice are more susceptible to excitotoxic injury and display a higher oxidative stress, highlighted by 8-Oxo-2'-deoxyguanosine increase and activation of defense mechanisms, as demonstrated by the increase of nuclear factor erythroid 2-related factor 2 (Nrf-2) in the nucleus. In Dach-SMOX astrocytes and neurons, an alteration of the phosphorylated and non-phosphorylated subunits of glutamate receptors increases the kainic acid response in these mice. Moreover, a decrease in excitatory amino acid transporters and an increase in the system x c - transporter, a Nrf-2 target, was observed. Sulfasalazine, a system x c - transporter inhibitor, was shown to revert the increased susceptibility of Dach-SMOX mice treated with kainic acid. We demonstrated that astrocytes play a crucial role in this process: neuronal spermine oxidase overexpression resulted in an alteration of glutamate excitability, in glutamate uptake and efflux in astrocytes involved in the synapse. Considering the involvement of oxidative stress in many neurodegenerative diseases, Dach-SMOX transgenic mouse can be considered as a suitable in vivo genetic model to study the involvement of spermine oxidase in excitotoxicity, which can be considered as a possible therapeutic target.

  7. Design, synthesis and biological evaluation of novel xanthine oxidase inhibitors bearing a 2-arylbenzo[b]furan scaffold.

    PubMed

    Tang, Hong-Jin; Li, Wei; Zhou, Mei; Peng, Li-Ying; Wang, Jin-Xin; Li, Jia-Huang; Chen, Jun

    2018-05-10

    Xanthine oxidase, which catalyzes the oxidative reaction of hypoxanthine and xanthine into uric acid, is a key enzyme to the pathogenesis of hyperuricemia and gout. In this study, for the purpose of discovering novel xanthine oxidase (XO) inhibitors, a series of 2-arylbenzo[b]furan derivatives (3a-3d, 4a-4o and 6a-6d) were designed and synthesized. All these compounds were evaluated their xanthine oxidase inhibitory and antioxidant activities by using in vitro enzymatic assay and cellular model. The results showed that a majority of the designed compounds exhibited potent xanthine oxidase inhibitory effects and antioxidant activities, and compound 4a emerged as the most potent xanthine oxidase inhibitor (IC 50  = 4.45 μM). Steady-state kinetic measurements of the inhibitor 4a with the bovine milk xanthine oxidase indicated a mixed type inhibition with 3.52 μM K i and 13.14 μM K is , respectively. The structure-activity relationship analyses have also been presented. Compound 4a exhibited the potent hypouricemic effect in the potassium oxonate-induced hyperuricemic mice model. A molecular docking study of compound 4a was performed to gain an insight into its binding mode with xanthine oxidase. These results highlight the identification of a new class of xanthine oxidase inhibitors that have potential to be more efficacious in treatment of gout. Copyright © 2018 Elsevier Masson SAS. All rights reserved.

  8. Alcohol use by alcoholics with and without a history of parental alcoholism.

    PubMed

    Worobec, T G; Turner, W M; O'Farrell, T J; Cutter, H S; Bayog, R D; Tsuang, M T

    1990-12-01

    The association between parental history of alcoholism and the nature of alcoholism was assessed using a more reliable measure of family history (Family Tree Questionnaire) and a more comprehensive inventory of alcoholism (Alcohol Use Inventory) than used in earlier studies. Parental alcoholism was associated with more severe alcoholism on most parameters of alcohol use (age of onset, quantity, frequency, preoccupation, and sustained use) and alcohol-related problems (social, vocational, physical, cognitive, and loss of control). The association between parental history of alcoholism and more severe alcoholism in the probands was independent of age of onset of alcoholism, current age, socioeconomic background, and marital status. Parental history positive (PH+) alcoholics were more reliant on alcohol to manage their moods but did not differ significantly from parental history negative (PH-) alcoholics in the use of alcohol to improve sociability or mental functioning or to cope with marital problems. Surprisingly, the degree of concern, guilt, and worry over the negative consequences of drinking was not significantly different for PH+ alcoholics although the negative consequences were clearly much more severe for this group. While the data are inconclusive about the reasons for more severe alcoholism in PH+ alcoholics, greater reliance on ethanol to manage moods and a relative insensitivity to negative consequences could theoretically account for the vulnerability to more severe alcoholism found in PH+ alcoholics.

  9. A decade of crystallization drops: Crystallization of the cbb3 cytochrome c oxidase from Pseudomonas stutzeri

    PubMed Central

    Buschmann, Sabine; Richers, Sebastian; Ermler, Ulrich; Michel, Hartmut

    2014-01-01

    The cbb3 cytochrome c oxidases are distant members of the superfamily of heme copper oxidases. These terminal oxidases couple O2 reduction with proton transport across the plasma membrane and, as a part of the respiratory chain, contribute to the generation of an electrochemical proton gradient. Compared with other structurally characterized members of the heme copper oxidases, the recently determined cbb3 oxidase structure at 3.2 Å resolution revealed significant differences in the electron supply system, the proton conducting pathways and the coupling of O2 reduction to proton translocation. In this paper, we present a detailed report on the key steps for structure determination. Improvement of the protein quality was achieved by optimization of the number of lipids attached to the protein as well as the separation of two cbb3 oxidase isoenzymes. The exchange of n-dodecyl-β-d-maltoside for a precisely defined mixture of two α-maltosides and decanoylsucrose as well as the choice of the crystallization method had a most profound impact on crystal quality. This report highlights problems frequently encountered in membrane protein crystallization and offers meaningful approaches to improve crystal quality. PMID:24488923

  10. A decade of crystallization drops: crystallization of the cbb3 cytochrome c oxidase from Pseudomonas stutzeri.

    PubMed

    Buschmann, Sabine; Richers, Sebastian; Ermler, Ulrich; Michel, Hartmut

    2014-04-01

    The cbb3 cytochrome c oxidases are distant members of the superfamily of heme copper oxidases. These terminal oxidases couple O2 reduction with proton transport across the plasma membrane and, as a part of the respiratory chain, contribute to the generation of an electrochemical proton gradient. Compared with other structurally characterized members of the heme copper oxidases, the recently determined cbb3 oxidase structure at 3.2 Å resolution revealed significant differences in the electron supply system, the proton conducting pathways and the coupling of O2 reduction to proton translocation. In this paper, we present a detailed report on the key steps for structure determination. Improvement of the protein quality was achieved by optimization of the number of lipids attached to the protein as well as the separation of two cbb3 oxidase isoenzymes. The exchange of n-dodecyl-β-D-maltoside for a precisely defined mixture of two α-maltosides and decanoylsucrose as well as the choice of the crystallization method had a most profound impact on crystal quality. This report highlights problems frequently encountered in membrane protein crystallization and offers meaningful approaches to improve crystal quality. © 2014 The Protein Society.

  11. Age-related ultrastructural and monoamine oxidase changes in the rat optic nerve.

    PubMed

    Taurone, S; Ripandelli, G; Minni, A; Lattanzi, R; Miglietta, S; Pepe, N; Fumagalli, L; Micera, A; Pastore, F S; Artico, M

    2016-01-01

    The aim of this paper is to study the morphology and the distribution of the monoamine oxidase enzymatic system in the optic nerve of 4 month-old Wistar (young) and 28 month-old Wistar (old) rats. The optic nerve was harvested from 20 young and old rats. The segment of optic nerve was divided longitudinally into two pieces, each 0.1 mm in length. The first piece was used for transmission electron microscopy. The second piece was stained with histochemical reaction for monoamine oxidase. The agerelated changes in the optic nerve of rats include micro-anatomical details, ultrastructure and monoamine oxidase histochemical staining. A strong decrease of the thin nerve fibers and a swelling of the thick ones can be observed in optic nerve fibers of old rats. Increased monoamine oxidase histochemical staining of the optic nerve of aged rats is well demonstrated. The increase of meningeal shealth and the decrease of thin nerve fibers of the optic nerve in old rats are well documented. Morphological, ultrastructural and histochemical changes observed in optic nerve fibers of the old rats show a close relation with aging.

  12. Abnormal kinetic behavior of cytochrome oxidase in a case of Leigh disease.

    PubMed Central

    Glerum, M; Robinson, B H; Spratt, C; Wilson, J; Patrick, D

    1987-01-01

    Cultured skin fibroblasts from a child with fatal lacticacidemia displayed an abnormally high lactate:pyruvate ratio of 77:1, compared with control values of 22:1-27:1. When protease-treated isolated mitochondria were used, activity of the respiratory-chain enzymes was found to be approximately 60% of normal, and adenosine triphosphate synthesis was found to be normal with all substrates tested. In mitochondria prepared by means of digitonin treatment, adenosine triphosphate synthesis was depressed with all substrates tested, suggesting a defect in the operation of the cytochrome oxidase complex. In disrupted whole cells from the patient, cytochrome oxidase activity was 56% of the activity in the control cell line with the lowest activity. In the presence of a twofold excess of oxidized cytochrome c, patient cells showed 31% of the activity in controls. Cytochrome oxidase activity in both sonicated whole-cell preparations and in sonicated mitochondria displayed abnormal kinetics with regard to the substrate-reduced cytochrome c, which was particularly evident in the presence of excess oxidized cytochrome c. We believe that kinetically abnormal cytochrome oxidase complex is responsible for the biochemical and clinical abnormalities present in this patient. PMID:2821802

  13. Genetics Home Reference: cytochrome c oxidase deficiency

    MedlinePlus

    ... are caused by mutations in genes found within nuclear DNA; however, in some rare instances, mutations in genes located within mtDNA cause this condition. The genes associated with cytochrome c oxidase deficiency are involved in energy production in mitochondria through a process called oxidative ...

  14. Associations Between MAOA-uVNTR Genotype, Maltreatment, MAOA Methylation, and Alcohol Consumption in Young Adult Males.

    PubMed

    Bendre, Megha; Comasco, Erika; Checknita, Dave; Tiihonen, Jari; Hodgins, Sheilagh; Nilsson, Kent W

    2018-03-01

    Epigenetic mechanisms are candidate moderators of the effect of maltreatment on brain and behavior. Interactions between maltreatment and the monoamine oxidase A upstream variable number tandem repeat genotype (MAOA-uVNTR) are associated with alcohol-related problems. However, presently it is not known whether DNA methylation moderates this association. The study focused on 53 young adult males and aimed to determine whether MAOA methylation moderated the association of alcohol-related problems with the interaction of MAOA-uVNTR and maltreatment, and whether alcohol consumption moderated the association of MAOA methylation with the interaction of MAOA-uVNTR and maltreatment. MAOA-uVNTR genotypes with ≤ 3 and > 3 repeats were categorized as short (S) and long (L), respectively. Data on maltreatment were obtained retrospectively, using self-reported questionnaires. DNA methylation of 16 candidate CpGs within part of the MAOA first exon and intron was assessed and grouped based on principal component analyses. Alcohol-related problems were assessed using the Alcohol Use Disorders Identification Test (AUDIT). Alcohol consumption was measured using AUDIT-C. Moderation effects were assessed and probed using the moderated moderation model and Johnson-Neyman's method, respectively. Carriers of the S allele, who experienced maltreatment and displayed lower Component 1 (mean of CpGs 13-16 in the first intron) MAOA methylation levels, reported higher AUDIT score in contrast to L-allele carriers. Carriers of the S allele, who reported higher AUDIT-C score and experienced maltreatment, displayed lower Component 3 (mean of CpGs 2-6 in the first exon) MAOA methylation levels than L-allele carriers. Intronic methylation moderated the association of alcohol-related problems with the interaction of MAOA-uVNTR and maltreatment. Alcohol consumption moderated the association of exonic methylation with the interaction of MAOA-uVNTR and maltreatment. These results suggest that

  15. Glycosylation site-targeted PEGylation of glucose oxidase retains native enzymatic activity.

    PubMed

    Ritter, Dustin W; Roberts, Jason R; McShane, Michael J

    2013-04-10

    Targeted PEGylation of glucose oxidase at its glycosylation sites was investigated to determine the effect on enzymatic activity, as well as the bioconjugate's potential in an optical biosensing assay. Methoxy-poly(ethylene glycol)-hydrazide (4.5kDa) was covalently coupled to periodate-oxidized glycosylation sites of glucose oxidase from Aspergillus niger. The bioconjugate was characterized using gel electrophoresis, liquid chromatography, mass spectrometry, and dynamic light scattering. Gel electrophoresis data showed that the PEGylation protocol resulted in a drastic increase (ca. 100kDa) in the apparent molecular mass of the protein subunit, with complete conversion to the bioconjugate; liquid chromatography data corroborated this large increase in molecular size. Mass spectrometry data proved that the extent of PEGylation was six poly(ethylene glycol) chains per glucose oxidase dimer. Dynamic light scattering data indicated the absence of higher-order oligomers in the PEGylated GOx sample. To assess stability, enzymatic activity assays were performed in triplicate at multiple time points over the course of 29 days in the absence of glucose, as well as before and after exposure to 5% w/v glucose for 24h. At a confidence level of 95%, the bioconjugate's performance was statistically equivalent to native glucose oxidase in terms of activity retention over the 29 day time period, as well as following the 24h glucose exposure. Finally, the bioconjugate was entrapped within a poly(2-hydroxyethyl methacrylate) hydrogel containing an oxygen-sensitive phosphor, and the construct was shown to respond approximately linearly with a 220±73% signal change (n=4, 95% confidence interval) over the physiologically-relevant glucose range (i.e., 0-400mg/dL); to our knowledge, this represents the first demonstration of PEGylated glucose oxidase incorporated into an optical biosensing assay. Copyright © 2013 Elsevier Inc. All rights reserved.

  16. NADPH Oxidase Inhibition Improves Neurological Outcomes in Surgically-Induced Brain Injury

    PubMed Central

    Lo, Wendy; Bravo, Thomas; Jadhav, Vikram; Zhang, John H.; Tang, Jiping

    2007-01-01

    Neurosurgical procedures can result in brain injury by various means including direct trauma, hemorrhage, retractor stretch, and electrocautery. This surgically-induced brain injury (SBI) can cause post-operative complications such as brain edema. By creating a mouse model of SBI, we tested whether NADPH oxidase, an important reactive oxygen species producing enzyme, is involved in SBI using transgenic mice lacking gp91phox subunit of NADPH oxidase (gp91phox KO) and apocynin, a specific inhibitor of NADPH oxidase. Neurological function and brain edema were evaluated at 24 hours post-SBI in gp91phox KO and wild-type littermates grouped into SBI and sham-surgery groups. Alternatively, mice were grouped into vehicle- and apocynin-treated (5mg/kg, i.p. 30 minutes before SBI) groups. Oxidative stress indicated by lipid peroxidation (LPO) was measured at 3 and 24 hours post SBI. The gp91phox KO mice, but not the apocynin-treated mice showed significantly improved neurological scores. Brain edema was observed in both gp91phox KO and wild-type groups after SBI; however, there was no significant difference between these two groups. Brain edema was also not affected by apocynin-pretreatment. LPO levels were significantly higher in SBI group in both gp91phox KO and wild-type groups as compared to sham group. A trend, although without statistical significance, was noted towards attenuation of LPO in the gp91phox KO animals as compared to wild-type group. LPO levels were significantly attenuated at 3 hours post-SBI by apocynin pretreatment but not at 24 hours post-SBI. These results suggest that chronic and acute inhibition of NADPH oxidase activity does not reduce brain edema after SBI. Long-term inhibition of NADPH oxidase, however improves neurological functions after SBI. PMID:17317004

  17. Gene cloning and heterologous expression of pyranose 2-oxidase from the brown-rot fungus, Gloeophyllum trabeum

    Treesearch

    Diane Dietrich; Casey Crooks

    2009-01-01

    A pyranose 2-oxidase gene from the brown-rot basidiomycete Gloeophyllum trabeum was isolated using homology-based degenerate PCR. The gene structure was determined and compared to that of several pyranose 2-oxidases cloned from white-rot fungi. The G. trabeum pyranose 2-oxidase gene consists of 16 coding exons with canonical promoter CAAT and TATA elements in the 5’UTR...

  18. What is addiction?

    PubMed

    Kranzler, Henry R; Li, Ting-Kai

    2008-01-01

    This issue of Alcohol Research & Health examines addiction to multiple substances--that is, combined dependence on alcohol and other drugs (AODs), including marijuana, cocaine, and opioids. It seems fitting, then, to begin the issue with a look at what constitutes "addiction." The Oxford English Dictionary (pp. 24-25) traces the term addiction to Roman law, under which addiction was a "formal giving over by sentence of court; hence, a dedication of person to a master." This notion of relinquishment of control by the addicted person is the central feature of many lay and professional definitions of the term. The study of addictive behavior crosses several disciplines, including, among others, behavioral neuroscience, epidemiology, genetics, molecular biology, pharmacology, psychology, psychiatry, and sociology. Articles in this issue examine aspects of AOD use disorders from the perspective of some of these varied disciplines.

  19. The GA5 locus of Arabidopsis thaliana encodes a multifunctional gibberellin 20-oxidase: molecular cloning and functional expression.

    PubMed

    Xu, Y L; Li, L; Wu, K; Peeters, A J; Gage, D A; Zeevaart, J A

    1995-07-03

    The biosynthesis of gibberellins (GAs) after GA12-aldehyde involves a series of oxidative steps that lead to the formation of bioactive GAs. Previously, a cDNA clone encoding a GA 20-oxidase [gibberellin, 2-oxoglutarate:oxygen oxidoreductase (20-hydroxylating, oxidizing), EC 1.14.11.-] was isolated by immunoscreening a cDNA library from liquid endosperm of pumpkin (Cucurbita maxima L.) with antibodies against partially purified GA 20-oxidase. Here, we report isolation of a genomic clone for GA 20-oxidase from a genomic library of the long-day species Arabidopsis thaliana Heynh., strain Columbia, by using the pumpkin cDNA clone as a heterologous probe. This genomic clone contains a GA 20-oxidase gene that consists of three exons and two introns. The three exons are 1131-bp long and encode 377 amino acid residues. A cDNA clone corresponding to the putative GA 20-oxidase genomic sequence was constructed with the reverse transcription-PCR method, and the identity of the cDNA clone was confirmed by analyzing the capability of the fusion protein expressed in Escherichia coli to convert GA53 to GA44 and GA19 to GA20. The Arabidopsis GA 20-oxidase shares 55% identity and > 80% similarity with the pumpkin GA 20-oxidase at the derived amino acid level. Both GA 20-oxidases share high homology with other 2-oxoglutarate-dependent dioxygenases (2-ODDs), but the highest homology was found between the two GA 20-oxidases. Mapping results indicated tight linkage between the cloned GA 20-oxidase and the GA5 locus of Arabidopsis. The ga5 semidwarf mutant contains a G-->A point mutation that inserts a translational stop codon in the protein-coding sequence, thus confirming that the GA5 locus encodes GA 20-oxidase. Expression of the GA5 gene in Ara-bidopsis leaves was enhanced after plants were transferred from short to long days; it was reduced by GA4 treatment, suggesting end-product repression in the GA biosynthetic pathway.

  20. Polyamine oxidase activity in rats treated with mitoguazone: specific and permanent decrease in thymus.

    PubMed

    Ferioli, M E; Armanni, A

    2003-01-01

    To extend the knowledge on the role of polyamine oxidase in thymus physiology, we evaluated the in vivo effect of the polyamine biosynthetic pathway inhibitor mitoguazone. The drug markedly and permanently decreased the enzyme activity in the organ, in which the level of putrescine also decreased at the later times observed. A byproduct of the reaction catalyzed by polyamine oxidase is hydrogen peroxide, a well known inducer of apoptosis. The decrease in polyamine oxidase activity, with the consequent decrease in hydrogen peroxide production, is correlated with a positive effect on thymus physiology. Since mitoguazone has been successfully employed in patients with AIDS-related diseases, in which the reconstitution of the immune function is a favorable prognostic index, we hypothesized that mitoguazone may have the thymus as target organ, and that the decrease in polyamine oxidase activity may have a role in the positive effect of the drug.

  1. Resveratrol protects vascular endothelial cells from high glucose-induced apoptosis through inhibition of NADPH oxidase activation-driven oxidative stress.

    PubMed

    Chen, Feng; Qian, Li-Hua; Deng, Bo; Liu, Zhi-Min; Zhao, Ying; Le, Ying-Ying

    2013-09-01

    Hyperglycemia-induced oxidative stress has been implicated in diabetic vascular complications in which NADPH oxidase is a major source of reactive oxygen species (ROS) generation. Resveratrol is a naturally occurring polyphenol, which has vasoprotective effects in diabetic animal models and inhibits high glucose (HG)-induced oxidative stress in endothelial cells. We aimed to examine whether HG-induced NADPH oxidase activation and ROS production contribute to glucotoxicity to endothelial cells and the effect of resveratrol on glucotoxicity. Using a murine brain microvascular endothelial cell line bEnd3, we found that NADPH oxidase inhibitor (apocynin) and resveratrol both inhibited HG-induced endothelial cell apoptosis. HG-induced elevation of NADPH oxidase activity and production of ROS were inhibited by apocynin, suggesting that HG induces endothelial cell apoptosis through NADPH oxidase-mediated ROS production. Mechanistic studies revealed that HG upregulated NADPH oxidase subunit Nox1 but not Nox2, Nox4, and p22(phox) expression through NF-κB activation, which resulted in elevation of NADPH oxidase activity and consequent ROS production. Resveratrol prevented HG-induced endothelial cell apoptosis through inhibiting HG-induced NF-κB activation, NADPH oxidase activity elevation, and ROS production. HG induces endothelial cell apoptosis through NF-κB/NADPH oxidase/ROS pathway, which was inhibited by resveratrol. Our findings provide new potential therapeutic targets against brain vascular complications of diabetes. © 2013 John Wiley & Sons Ltd.

  2. Oxoferryl-Porphyrin Radical Catalytic Intermediate in Cytochrome bd Oxidases Protects Cells from Formation of Reactive Oxygen Species*

    PubMed Central

    Paulus, Angela; Rossius, Sebastiaan Gijsbertus Hendrik; Dijk, Madelon; de Vries, Simon

    2012-01-01

    The quinol-linked cytochrome bd oxidases are terminal oxidases in respiration. These oxidases harbor a low spin heme b558 that donates electrons to a binuclear heme b595/heme d center. The reaction with O2 and subsequent catalytic steps of the Escherichia coli cytochrome bd-I oxidase were investigated by means of ultra-fast freeze-quench trapping followed by EPR and UV-visible spectroscopy. After the initial binding of O2, the O–O bond is heterolytically cleaved to yield a kinetically competent heme d oxoferryl porphyrin π-cation radical intermediate (compound I) magnetically interacting with heme b595. Compound I accumulates to 0.75–0.85 per enzyme in agreement with its much higher rate of formation (∼20,000 s−1) compared with its rate of decay (∼1,900 s−1). Compound I is next converted to a short lived heme d oxoferryl intermediate (compound II) in a phase kinetically matched to the oxidation of heme b558 before completion of the reaction. The results indicate that cytochrome bd oxidases like the heme-copper oxidases break the O–O bond in a single four-electron transfer without a peroxide intermediate. However, in cytochrome bd oxidases, the fourth electron is donated by the porphyrin moiety rather than by a nearby amino acid. The production of reactive oxygen species by the cytochrome bd oxidase was below the detection level of 1 per 1000 turnovers. We propose that the two classes of terminal oxidases have mechanistically converged to enzymes in which the O–O bond is broken in a single four-electron transfer reaction to safeguard the cell from the formation of reactive oxygen species. PMID:22287551

  3. Effect of voluntary alcohol consumption on Maoa expression in the mesocorticolimbic brain of adult male rats previously exposed to prolonged maternal separation

    PubMed Central

    Bendre, M; Comasco, E; Nylander, I; Nilsson, K W

    2015-01-01

    Discordant associations between monoamine oxidase A (MAOA) genotype and high alcohol drinking have been reported in human and non-human primates. Environmental influences likely moderate genetic susceptibility. The biological basis for this interplay remains elusive, and inconsistencies call for translational studies in which conditions can be controlled and brain tissue is accessible. The present study investigated whether early life stress and subsequent adult episodic alcohol consumption affect Maoa expression in stress- and reward-related brain regions in the rat. Outbred Wistar rats were exposed to rearing conditions associated with stress (prolonged maternal separation) or no stress during early life, and given free choice between alcohol and/or water in adulthood. Transcript levels of Maoa were assessed in the ventral tegmental area, nucleus accumbens (NAc), medial prefrontal cortex, cingulate cortex, amygdala and dorsal striatum (DS). Blood was collected to assess corticosterone levels. After alcohol consumption, lower blood corticosterone and Maoa expression in the NAc and DS were found in rats exposed to early life stress compared with control rats. An interaction between early life stress and voluntary alcohol intake was found in the NAc. Alcohol intake before death correlated negatively with Maoa expression in DS in high alcohol-drinking rats exposed to early life stress. Maoa expression is sensitive to adulthood voluntary alcohol consumption in the presence of early life stress in outbred rats. These findings add knowledge of the molecular basis of the previously reported associations between early life stress, MAOA and susceptibility to alcohol misuse. PMID:26645625

  4. Structure-Based Alteration of Substrate Specificity and Catalytic Activity of Sulfite Oxidase from Sulfite Oxidation to Nitrate Reduction

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

    Qiu, James A.; Wilson, Heather L.; Rajagopalan, K.V.

    Eukaryotic sulfite oxidase is a dimeric protein that contains the molybdenum cofactor and catalyzes the metabolically essential conversion of sulfite to sulfate as the terminal step in the metabolism of cysteine and methionine. Nitrate reductase is an evolutionarily related molybdoprotein in lower organisms that is essential for growth on nitrate. In this study, we describe human and chicken sulfite oxidase variants in which the active site has been modified to alter substrate specificity and activity from sulfite oxidation to nitrate reduction. On the basis of sequence alignments and the known crystal structure of chicken sulfite oxidase, two residues are conservedmore » in nitrate reductases that align with residues in the active site of sulfite oxidase. On the basis of the crystal structure of yeast nitrate reductase, both positions were mutated in human sulfite oxidase and chicken sulfite oxidase. The resulting double-mutant variants demonstrated a marked decrease in sulfite oxidase activity but gained nitrate reductase activity. An additional methionine residue in the active site was proposed to be important in nitrate catalysis, and therefore, the triple variant was also produced. The nitrate reducing ability of the human sulfite oxidase triple mutant was nearly 3-fold greater than that of the double mutant. To obtain detailed structural data for the active site of these variants, we introduced the analogous mutations into chicken sulfite oxidase to perform crystallographic analysis. The crystal structures of the Mo domains of the double and triple mutants were determined to 2.4 and 2.1 {angstrom} resolution, respectively.« less

  5. Role of Valine 464 in the Flavin Oxidation Reaction Catalyzed by Choline Oxidase

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

    Finnegan, Steffan; Agniswamy, Johnson; Weber, Irene T.

    2010-11-03

    The oxidation of reduced flavin cofactors by oxygen is a very important reaction that is central to the chemical versatility of hundreds of flavoproteins classified as monooxygenases and oxidases. These enzymes are characterized by bimolecular rate constants {ge} 10{sup 5} M{sup -1} s{sup -1} and produce water and hydrogen peroxide, respectively. A hydrophobic cavity close to the reactive flavin C(4a) atom has been previously identified in the 3D structure of monooxygenases but not in flavoprotein oxidases. In the present study, we have investigated by X-ray crystallography, mutagenesis, steady-state, and rapid reaction approaches the role of Val464, which is <6 {angstrom}more » from the flavin C(4a) atom in choline oxidase. The 3D structure of the Val464Ala enzyme was essentially identical to that of the wild-type enzyme as shown by X-ray crystallography. Time-resolved anaerobic substrate reduction of the enzymes showed that replacement of Val464 with alanine or threonine did not affect the reductive half-reaction. Steady-state and rapid kinetics as well as enzyme-monitored turnovers indicated that the oxidative half-reaction in the Ala464 and Thr464 enzymes was decreased by 50-fold with respect to the wild-type enzyme. We propose that the side chain of Val464 in choline oxidase provides a nonpolar site that is required to guide oxygen in proximity of the C(4a) atom of the flavin, where it will subsequently react via electrostatic catalysis. Visual analysis of available structures suggests that analogous nonpolar sites are likely present in most flavoprotein oxidases. Mechanistic considerations provide rationalization for the differences between sites in monooxygenases and oxidases.« less

  6. Hypouricaemic action of mangiferin results from metabolite norathyriol via inhibiting xanthine oxidase activity.

    PubMed

    Niu, Yanfen; Liu, Jia; Liu, Hai-Yang; Gao, Li-Hui; Feng, Guo-Hua; Liu, Xu; Li, Ling

    2016-09-01

    Context Mangiferin has been reported to possess a potential hypouricaemic effect. However, the pharmacokinetic studies in rats showed that its oral bioavailability was only 1.2%, suggesting that mangiferin metabolites might exert the action. Objective The hypouricaemic effect and the xanthine oxidase inhibition of mangiferin and norathyriol, a mangiferin metabolite, were investigated. Inhibition of norathyriol analogues (compounds 3-9) toward xanthine oxidase was also evaluated. Materials and methods For a dose-dependent study, mangiferin (1.5-6.0 mg/kg) and norathyriol (0.92-3.7 mg/kg) were administered intragastrically to mice twice daily for five times. For a time-course study, mice received mangiferin and norathyriol both at a single dose of 7.1 μmol/kg. In vitro, inhibition of test compounds (2.4-2.4 mM) against xanthine oxidase activity was evaluated by the spectrophotometrical method. The inhibition type was identified from Lineweaver-Burk plots. Results Norathyriol (0.92, 1.85 and 3.7 mg/kg) dose dependently decreased the serum urate levels by 27.0, 33.6 and 37.4%, respectively. The action was more potent than that of mangiferin at the low dose, but was equivalent at the higher doses. Additionally, the hypouricaemic action of them exhibited a time dependence. In vitro, norathyriol markedly inhibited the xanthine oxidase activities, with the IC50 value of 44.6 μM, but mangiferin did not. The kinetic studies showed that norathyriol was an uncompetitive inhibitor by Lineweaver-Burk plots. The structure-activity relationships exhibited that three hydroxyl groups in norathyriol at the C-1, C-3 and C-6 positions were essential for maintaining xanthine oxidase inhibition. Discussion and conclusion Norathyriol was responsible for the hypouricaemic effect of mangiferin via inhibiting xanthine oxidase activity.

  7. Biochemical Conservation and Evolution of Germacrene A Oxidase in Asteraceae*

    PubMed Central

    Nguyen, Don Trinh; Göpfert, Jens Christian; Ikezawa, Nobuhiro; MacNevin, Gillian; Kathiresan, Meena; Conrad, Jürgen; Spring, Otmar; Ro, Dae-Kyun

    2010-01-01

    Sesquiterpene lactones are characteristic natural products in Asteraceae, which constitutes ∼8% of all plant species. Despite their physiological and pharmaceutical importance, the biochemistry and evolution of sesquiterpene lactones remain unexplored. Here we show that germacrene A oxidase (GAO), evolutionarily conserved in all major subfamilies of Asteraceae, catalyzes three consecutive oxidations of germacrene A to yield germacrene A acid. Furthermore, it is also capable of oxidizing non-natural substrate amorphadiene. Co-expression of lettuce GAO with germacrene synthase in engineered yeast synthesized aberrant products, costic acids and ilicic acid, in an acidic condition. However, cultivation in a neutral condition allowed the de novo synthesis of a single novel compound that was identified as germacrene A acid by gas and liquid chromatography and NMR analyses. To trace the evolutionary lineage of GAO in Asteraceae, homologous genes were further isolated from the representative species of three major subfamilies of Asteraceae (sunflower, chicory, and costus from Asteroideae, Cichorioideae, and Carduoideae, respectively) and also from the phylogenetically basal species, Barnadesia spinosa, from Barnadesioideae. The recombinant GAOs from these genes clearly showed germacrene A oxidase activities, suggesting that GAO activity is widely conserved in Asteraceae including the basal lineage. All GAOs could catalyze the three-step oxidation of non-natural substrate amorphadiene to artemisinic acid, whereas amorphadiene oxidase diverged from GAO displayed negligible activity for germacrene A oxidation. The observed amorphadiene oxidase activity in GAOs suggests that the catalytic plasticity is embedded in ancestral GAO enzymes that may contribute to the chemical and catalytic diversity in nature. PMID:20351109

  8. Hypoglycemic neuronal death is triggered by glucose reperfusion and activation of neuronal NADPH oxidase

    PubMed Central

    Suh, Sang Won; Gum, Elizabeth T.; Hamby, Aaron M.; Chan, Pak H.; Swanson, Raymond A.

    2007-01-01

    Hypoglycemic coma and brain injury are potential complications of insulin therapy. Certain neurons in the hippocampus and cerebral cortex are uniquely vulnerable to hypoglycemic cell death, and oxidative stress is a key event in this cell death process. Here we show that hypoglycemia-induced oxidative stress and neuronal death are attributable primarily to the activation of neuronal NADPH oxidase during glucose reperfusion. Superoxide production and neuronal death were blocked by the NADPH oxidase inhibitor apocynin in both cell culture and in vivo models of insulin-induced hypoglycemia. Superoxide production and neuronal death were also blocked in studies using mice or cultured neurons deficient in the p47phox subunit of NADPH oxidase. Chelation of zinc with calcium disodium EDTA blocked both the assembly of the neuronal NADPH oxidase complex and superoxide production. Inhibition of the hexose monophosphate shunt, which utilizes glucose to regenerate NADPH, also prevented superoxide formation and neuronal death, suggesting a mechanism linking glucose reperfusion to superoxide formation. Moreover, the degree of superoxide production and neuronal death increased with increasing glucose concentrations during the reperfusion period. These results suggest that high blood glucose concentrations following hypoglycemic coma can initiate neuronal death by a mechanism involving extracellular zinc release and activation of neuronal NADPH oxidase. PMID:17404617

  9. Purification and partial amino-acid sequence of gibberellin 20-oxidase from Cucurbita maxima L. endosperm.

    PubMed

    Lange, T

    1994-01-01

    Gibberellin (GA) 20-oxidase was purified to apparent homogeneity from Cucurbita maxima endosperm by fractionated ammonium-sulphate precipitation, gel-filtration chromatography and anion-exchange and hydrophobic-interaction high-performance liquid chromatography (HPLC). Average purification after the last step was 55-fold with 3.9% of the activity recovered. The purest single fraction was enriched 101-fold with 0.2% overall recovery. Apparent relative molecular mass of the enzyme was 45 kDa, as determined by gel-filtration HPLC and sodium dodecyl sulphate-polyacrylamide gel electrophoresis, indicating that GA 20-oxidase is probably a monomeric enzyme. The purified enzyme degraded on two-dimensional gel electrophoresis, giving two protein spots: a major one corresponding to a molecular mass of 30 kDa and a minor one at 45 kDa. The isoelectric point for both was 5.4. The amino-acid sequences of the amino-terminus of the purified enzyme and of two peptides from a tryptic digest were determined. The purified enzyme catalysed the sequential conversion of [14C]GA12 to [14C]GA15, [14C]GA24 and [14C]GA25, showing that carbon atom 20 was oxidised to the corresponding alcohol, aldehyde and carboxylic acid in three consecutive reactions. [14C]Gibberellin A53 was similarly converted to [14C]GA44, [14C]GA19, [14C]GA17 and small amounts of a fourth product, which was preliminarily identified as [14C]GA20, a C19-gibberellin. All GAs except [14C]GA20 were identified by combined gas chromatography-mass spectrometry. The cofactor requirements in the absence of dithiothreitol were essentially as in its presence (Lange et al., Planta 195, 98-107, 1994), except that ascorbate was essential for enzyme activity and the optimal concentration of catalase was lower.

  10. Overexpression of 20-Oxidase Confers a Gibberellin-Overproduction Phenotype in Arabidopsis

    PubMed Central

    Huang, Shihshieh; Raman, Anuradha S.; Ream, Joel E.; Fujiwara, Hideji; Cerny, R. Eric; Brown, Sherri M.

    1998-01-01

    In the gibberellin (GA) biosynthesis pathway, 20-oxidase catalyzes the oxidation and elimination of carbon-20 to give rise to C19-GAs. All bioactive GAs are C19-GAs. We have overexpressed a cDNA encoding 20-oxidase isolated from Arabidopsis seedlings in transgenic Arabidopsis plants. These transgenic plants display a phenotype that may be attributed to the overproduction of GA. The phenotype includes a longer hypocotyl, lighter-green leaves, increased stem elongation, earlier flowering, and decreased seed dormancy. However, the fertility of the transgenic plants is not affected. Increased levels of endogenous GA1, GA9, and GA20 were detected in seedlings of the transgenic line examined. GA4, which is thought to be the predominantly active GA in Arabidopsis, was not present at increased levels in this line. These results suggest that the overexpression of this 20-oxidase increases the levels of some endogenous GAs in transgenic seedlings, which causes the GA-overproduction phenotype. PMID:9808721

  11. Conformational lock and dissociative thermal inactivation of lentil seedling amine oxidase.

    PubMed

    Moosavi-Nejad, S Zahra; Moosavi-Movahedi, Ali-Akbar; Rezaei-Tavirani, Mostafa; Floris, Giovanni; Medda, Rosaria

    2003-03-31

    The kinetics of thermal inactivation of copper-containing amine oxidase from lentil seedlings were studied in a 100 mM potassium phosphate buffer, pH 7, using putrescine as the substrate. The temperature range was between 47-60 degrees C. The thermal inactivation curves were not linear at 52 and 57 degrees C; three linear phases were shown. The first phase gave some information about the number of dimeric forms of the enzyme that were induced by the higher temperatures using the "conformational lock" pertaining theory to oligomeric enzyme. The "conformational lock" caused two additional dimeric forms of the enzyme when the temperature increased to 57 degrees C. The second and third phases were interpreted according to a dissociative thermal inactivation model. These phases showed that lentil amine oxidase was reversibly-dissociated before the irreversible thermal inactivation. Although lentil amine oxidase is not a thermostable enzyme, its dimeric structure can form "conformational lock," conferring a structural tolerance to the enzyme against heat stress.

  12. A Mycobacterium tuberculosis cytochrome bd oxidase mutant is hypersensitive to bedaquiline.

    PubMed

    Berney, Michael; Hartman, Travis E; Jacobs, William R

    2014-07-15

    The new medicinal compound bedaquiline (BDQ) kills Mycobacterium tuberculosis by inhibiting F1Fo-ATP synthase. BDQ is bacteriostatic for 4 to 7 days and kills relatively slowly compared to other frontline tuberculosis (TB) drugs. Here we show that killing with BDQ can be improved significantly by inhibiting cytochrome bd oxidase, a non-proton-pumping terminal oxidase. BDQ was instantly bactericidal against a cytochrome bd oxidase null mutant of M. tuberculosis, and the rate of killing was increased by more than 50%. We propose that this exclusively bacterial enzyme should be a high-priority target for new drug discovery. Importance: A major drawback of current TB chemotherapy is its long duration. New drug regimens with rapid killing kinetics are desperately needed. Our study demonstrates that inhibition of a nonessential bacterial enzyme greatly improves the efficacy of the latest TB drug bedaquiline and emphasizes that screening for compounds with synergistic killing mechanisms is a promising strategy. Copyright © 2014 Berney et al.

  13. The glucose oxidase-peroxidase assay for glucose

    USDA-ARS?s Scientific Manuscript database

    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...

  14. Expanding the substrate scope of chitooligosaccharide oxidase from Fusarium graminearum by structure-inspired mutagenesis.

    PubMed

    Ferrari, Alessandro R; Lee, Misun; Fraaije, Marco W

    2015-06-01

    Chitooligosaccharide oxidase from Fusarium graminearum (ChitO) oxidizes N-acetyl-D-glucosamine (GlcNAc) and its oligomers with high efficiency at the C1-hydroxyl moiety while it shows poor or no activity with other carbohydrates. By sequence and structural comparison with other known carbohydrate oxidases (glucooligosaccharide oxidase from Acremonium strictum and lactose oxidase from Microdochium nivale) eleven mutants were designed to redirect the catalytic scope of ChitO for improved oxidation of lactose, cellobiose and maltose. The catalytic properties of the most interesting mutants were further improved by combining single mutations. This has resulted in the creation of a set of ChitO variants that display totally different substrate tolerances. One ChitO variant shows a dramatic improvement in catalytic efficiency towards oxidation of glucose, cellobiose, lactose, and maltose. We also describe a ChitO variant with the highest catalytic efficiency in GlcNAc oxidation so far reported in the literature. © 2015 Wiley Periodicals, Inc.

  15. Regulation of superoxide anion production by NADPH oxidase in monocytes/macrophages: contributions to atherosclerosis.

    PubMed

    Cathcart, Martha K

    2004-01-01

    Monocyte extravasation into the vessel wall has been shown to be a critical step in the development of atherosclerosis. Upon activation, monocytes produce a burst of superoxide anion due to activation of the NADPH oxidase enzyme complex. Monocyte-derived superoxide anion contributes to oxidant stress in inflammatory sites, is required for monocyte-mediated LDL oxidation, and alters basic cell functions such as adhesion and proliferation. We hypothesize that monocyte-derived superoxide anion production contributes to atherosclerotic lesion formation. In this brief review, we summarize our current understanding of the signal transduction pathways regulating NADPH oxidase activation and related superoxide anion production in activated human monocytes. Novel pathways are identified that may serve as future targets for therapeutic intervention in this pathogenic process. The contributions of superoxide anion and NADPH oxidase to atherogenesis are discussed. Future experiments are needed to clarify the exact role of NADPH oxidase-derived superoxide anion in atherogenesis, particularly that derived from monocytes.

  16. Interrupted reperfusion reduces the activation of NADPH oxidase after cerebral I/R injury.

    PubMed

    Shen, Jia; Bai, Xiao-Yin; Qin, Yuan; Jin, Wei-Wei; Zhou, Jing-Yin; Zhou, Ji-Ping; Yan, Ying-Gang; Wang, Qiong; Bruce, Iain C; Chen, Jiang-Hua; Xia, Qiang

    2011-06-15

    Interrupted reperfusion reduces ischemia/reperfusion (I/R) injury. This study was designed to determine whether NADPH oxidase participates in the neural protection against global I/R injury after interrupted reperfusion. Mice were randomly divided into five groups: sham (sham-operated), I/R (20-min global I/R), RR (I/R+interrupted reperfusion), Apo (I/R+apocynin administration), and RR+Apo. Behavioral tests (pole test, beam walking, and Morris water maze) and Nissl staining were undertaken in all five groups; superoxide levels, expression of gp91(phox) and p47(phox), p47(phox) translocation, and Rac1 activation were measured in the sham, I/R, and RR groups. The motor coordination, bradykinesia, and spatial learning and memory, as well as the neuron survival rates, were better in the RR, Apo, and RR+Apo groups than in the I/R group. The NADPH oxidase-dependent superoxide levels, p47(phox) and gp91(phox) expression, p47(phox) translocation, and Rac1 activation were lower in the RR group than in the I/R group. In conclusion, the neural protective effect of interrupted reperfusion is at least partly mediated by decreasing the expression and assembly of NADPH oxidase and the levels of NADPH oxidase-derived superoxide. The most striking reduction Rac1-GTP in the RR group suggests that interrupted reperfusion also acts on the activation of assembled NADPH oxidase by reducing the availability of Rac1-GTP. Copyright © 2011 Elsevier Inc. All rights reserved.

  17. Absence of Proton Channels in COS-7 Cells Expressing Functional NADPH Oxidase Components

    PubMed Central

    Morgan, Deri; Cherny, Vladimir V.; Price, Marianne O.; Dinauer, Mary C.; DeCoursey, Thomas E.

    2002-01-01

    Nicotinamide adenine dinucleotide phosphate (NADPH) oxidase is an enzyme of phagocytes that produces bactericidal superoxide anion (O2 −) via an electrogenic process. Proton efflux compensates for the charge movement across the cell membrane. The proton channel responsible for the H+ efflux was thought to be contained within the gp91phox subunit of NADPH oxidase, but recent data do not support this idea (DeCoursey, T.E., V.V. Cherny, D. Morgan, B.Z. Katz, and M.C. Dinauer. 2001. J. Biol. Chem. 276:36063–36066). In this study, we investigated electrophysiological properties and superoxide production of COS-7 cells transfected with all NADPH oxidase components required for enzyme function (COSphox). The 7D5 antibody, which detects an extracellular epitope of the gp91phox protein, labeled 96–98% of COSphox cells. NADPH oxidase was functional because COSphox (but not COSWT) cells stimulated by phorbol myristate acetate (PMA) or arachidonic acid (AA) produced superoxide anion. No proton currents were detected in either wild-type COS-7 cells (COSWT) or COSphox cells studied at pHo 7.0 and pHi 5.5 or 7.0. Anion currents that decayed at voltages positive to 40 mV were the only currents observed. PMA or AA did not elicit detectable H+ current in COSWT or COSphox cells. Therefore, gp91phox does not function as a proton channel in unstimulated cells or in activated cells with a demonstrably functional oxidase. PMID:12034764

  18. Myeloperoxidase amplified high glucose-induced endothelial dysfunction in vasculature: Role of NADPH oxidase and hypochlorous acid.

    PubMed

    Tian, Rong; Ding, Yun; Peng, Yi-Yuan; Lu, Naihao

    2017-03-11

    Nicotinamide adenine dinucleotide phosphate (NADPH) oxidase-derived reactive oxygen species (ROS) such as superoxide and hydrogen peroxide (H 2 O 2 ), have emerged as important molecules in the pathogenesis of diabetic endothelial dysfunction. Additionally, neutrophils-derived myeloperoxidase (MPO) and MPO-catalyzed hypochlorous acid (HOCl) play important roles in the vascular injury. However, it is unknown whether MPO can use vascular-derived ROS to induce diabetic endothelial dysfunction. In the present study, we demonstrated that NADPH oxidase was the main source of ROS formation in high glucose-cultured human umbilical vein endothelial cells (HUVECs), and played a critical role in high glucose-induced endothelial dysfunction such as cell apoptosis, loss of cell viability and reduction of nitric oxide (NO). However, the addition of MPO could amplify the high glucose-induced endothelial dysfunction which was inhibited by the presence of apocynin (NADPH oxidase inhibitor), catalase (H 2 O 2 scavenger), or methionine (HOCl scavenger), demonstrating the contribution of NADPH oxidase-H 2 O 2 -MPO-HOCl pathway in the MPO/high glucose-induced vascular injury. In high glucose-incubated rat aortas, MPO also exacerbated the NADPH oxidase-induced impairment of endothelium-dependent relaxation. Consistent with these in vitro data, in diabetic rat aortas, both MPO expresion and NADPH oxidase activity were increased while the endothelial function was simultaneously impaired. The results suggested that vascular-bound MPO could amplify high glucose-induced vascular injury in diabetes. MPO-NADPH oxidase-HOCl may represent an important pathogenic pathway in diabetic vascular diseases. Copyright © 2017 Elsevier Inc. All rights reserved.

  19. Mannitol oxidase and polyol dehydrogenases in the digestive gland of gastropods: Correlations with phylogeny and diet

    PubMed Central

    Amaral-de-Carvalho, Diogo; Oliveira, Elsa; Alves, Ângela; Costa, Vítor; Calado, Gonçalo

    2018-01-01

    Mannitol oxidase and polyol dehydrogenases are enzymes that convert polyalcohols into sugars. Mannitol oxidase was previously investigated in terrestrial snails and slugs, being also present in a few aquatic gastropods. However, the overall distribution of this enzyme in the Gastropoda was not known. Polyol dehydrogenases are also poorly studied in gastropods and other mollusks. In this study, polyalcohol oxidase and dehydrogenase activities were assayed in the digestive gland of 26 species of gastropods, representing the clades Patellogastropoda, Neritimorpha, Vetigastropoda, Caenogastropoda and Heterobranchia. Marine, freshwater and terrestrial species, including herbivores and carnivores were analyzed. Ultrastructural observations were undertake in species possessing mannitol oxidase, in order to investigate the correlation between this enzyme and the presence of tubular structures known to be associated with it. Mannitol oxidase activity was detected in the digestive gland of herbivores from the clades Caenogastropoda and Heterobranchia, but not in any carnivores or in herbivores from the clades Patellogastropoda, Neritimorpha and Vetigastropoda. In most of the species used in this study, dehydrogenase activities were detected using both D-mannitol and D-sorbitol as substrates. Nevertheless, in some carnivores these activities were not detected with both polyalcohols. Ultrastructural observations revealed tubular structures in digestive gland cells of some species having mannitol oxidase activity, but they were not observed in others. Based on our results, we suggest that mannitol oxidase first occurred in a herbivorous or omnivorous ancestor of Apogastropoda, the clade formed by caenogastropods and heterobranchs, being subsequently lost in those species that shifted towards a carnivorous diet. PMID:29529078

  20. Exploiting the synthetic lethality between terminal respiratory oxidases to kill Mycobacterium tuberculosis and clear host infection

    PubMed Central

    Kalia, Nitin P.; Hasenoehrl, Erik J.; Ab Rahman, Nurlilah B.; Koh, Vanessa H.; Ang, Michelle L. T.; Sajorda, Dannah R.; Hards, Kiel; Grüber, Gerhard; Alonso, Sylvie; Cook, Gregory M.; Berney, Michael; Pethe, Kevin

    2017-01-01

    The recent discovery of small molecules targeting the cytochrome bc1:aa3 in Mycobacterium tuberculosis triggered interest in the terminal respiratory oxidases for antituberculosis drug development. The mycobacterial cytochrome bc1:aa3 consists of a menaquinone:cytochrome c reductase (bc1) and a cytochrome aa3-type oxidase. The clinical-stage drug candidate Q203 interferes with the function of the subunit b of the menaquinone:cytochrome c reductase. Despite the affinity of Q203 for the bc1:aa3 complex, the drug is only bacteriostatic and does not kill drug-tolerant persisters. This raises the possibility that the alternate terminal bd-type oxidase (cytochrome bd oxidase) is capable of maintaining a membrane potential and menaquinol oxidation in the presence of Q203. Here, we show that the electron flow through the cytochrome bd oxidase is sufficient to maintain respiration and ATP synthesis at a level high enough to protect M. tuberculosis from Q203-induced bacterial death. Upon genetic deletion of the cytochrome bd oxidase-encoding genes cydAB, Q203 inhibited mycobacterial respiration completely, became bactericidal, killed drug-tolerant mycobacterial persisters, and rapidly cleared M. tuberculosis infection in vivo. These results indicate a synthetic lethal interaction between the two terminal respiratory oxidases that can be exploited for anti-TB drug development. Our findings should be considered in the clinical development of drugs targeting the cytochrome bc1:aa3, as well as for the development of a drug combination targeting oxidative phosphorylation in M. tuberculosis. PMID:28652330

  1. [Out of addictions: Alcohol, or alcohol to alcohol].

    PubMed

    Simmat-Durand, L; Vellut, N; Lejeune, C; Jauffret-Roustide, M; Mougel, S; Michel, L; Planche, M

    2017-08-01

    Pathways from alcoholism to recovery are documented; less often are those from drug addiction to alcoholism. Biographical approaches allow analyzing how people change their uses and talk about their trajectories of recovery. Three hundred and forty-one people (34% women) in the Paris area were questioned on their trajectories with a biographical questionnaire. Some open questions were aimed to understand the connection they made between events in their lives, how recovered they felt and what they considered strengths or obstacles. All the participants had stopped at least one product. Their mean age was 43, and 26% were over 50. How can the differences between one substance addicts and dual abusers be explained? Can we hypothesize a better result for the patients with a single dependence to alcohol in their lives for the following two reasons? (1) They could really be taken in charge for their alcoholism whereas the dual abusers mostly receive cared for their illicit drug problems with an under estimation of their problem with alcohol. In this case, they turn to alcohol after weaning themselves from their drug dependence so as to return to a social consumption, especially when they are given an opiate treatment. (2) Conversely could we suggest that the dual substance abusers had different trajectories from their childhood (more adverse events, more social difficulties, mental health problems), and that this accumulation explains their skipping from one substance or behaviour to another without any real recovery for decades? All respondents were polydrug users. Eighty-two had been dependent mainly on alcohol. One hundred and twenty-one people had been drug addicts (mostly heroin), which they had stopped on average ten years before the survey. The last group included 138 persons who had been heroin or cocaine addicts and alcoholics in their lives, a third of whom had been dependent on alcohol before their drug addiction (35%), a tenth on both at the same time (10

  2. The Intimate and Controversial Relationship between Voltage Gated Proton Channels and the Phagocyte NADPH Oxidase

    PubMed Central

    DeCoursey, Thomas E.

    2016-01-01

    Summary One of the most fascinating and exciting periods in my scientific career entailed dissecting the symbiotic relationship between two membrane transporters, the NADPH oxidase complex and voltage gated proton channels (HV1). By the time I entered this field, there had already been substantial progress toward understanding NADPH oxidase, but HV1 were known only to a tiny handful of cognoscenti around the world. Having identified the first proton currents in mammalian cells in 1991, I needed to find a clear function for these molecules if the work was to become fundable. The then-recent discoveries of Henderson, Chappell, and colleagues in 1987–1988 that led them to hypothesize interactions of both molecules during the respiratory burst of phagocytes provided an excellent opportunity. In a nutshell, both transporters function by moving electrical charge across the membrane: NADPH oxidase moves electrons and HV1 moves protons. The consequences of electrogenic NADPH oxidase activity on both membrane potential and pH strongly self-limit this enzyme. Fortunately, both consequences specifically activate HV1, and HV1 activity counteracts both consequences, a kind of yin-yang relationship. Notwithstanding a decade starting in 1995 when many believed the opposite, these are two separate molecules that function independently despite their being functionally interdependent in phagocytes. The relationship between NADPH oxidase and HV1 has become a paradigm that somewhat surprisingly has now extended well beyond the phagocyte NADPH oxidase -- an industrial strength producer of reactive oxygen species (ROS) -- to myriad other cells that produce orders of magnitude less ROS for signaling purposes. These cells with their seven NADPH oxidase (NOX) isoforms provide a vast realm of mechanistic obscurity that will occupy future studies for years to come. PMID:27558336

  3. The intimate and controversial relationship between voltage-gated proton channels and the phagocyte NADPH oxidase.

    PubMed

    DeCoursey, Thomas E

    2016-09-01

    One of the most fascinating and exciting periods in my scientific career entailed dissecting the symbiotic relationship between two membrane transporters, the Nicotinamide adenine dinucleotide phosphate reduced form (NADPH) oxidase complex and voltage-gated proton channels (HV 1). By the time I entered this field, there had already been substantial progress toward understanding NADPH oxidase, but HV 1 were known only to a tiny handful of cognoscenti around the world. Having identified the first proton currents in mammalian cells in 1991, I needed to find a clear function for these molecules if the work was to become fundable. The then-recent discoveries of Henderson, Chappell, and colleagues in 1987-1988 that led them to hypothesize interactions of both molecules during the respiratory burst of phagocytes provided an excellent opportunity. In a nutshell, both transporters function by moving electrical charge across the membrane: NADPH oxidase moves electrons and HV 1 moves protons. The consequences of electrogenic NADPH oxidase activity on both membrane potential and pH strongly self-limit this enzyme. Fortunately, both consequences specifically activate HV 1, and HV 1 activity counteracts both consequences, a kind of yin-yang relationship. Notwithstanding a decade starting in 1995 when many believed the opposite, these are two separate molecules that function independently despite their being functionally interdependent in phagocytes. The relationship between NADPH oxidase and HV 1 has become a paradigm that somewhat surprisingly has now extended well beyond the phagocyte NADPH oxidase - an industrial strength producer of reactive oxygen species (ROS) - to myriad other cells that produce orders of magnitude less ROS for signaling purposes. These cells with their seven NADPH oxidase (NOX) isoforms provide a vast realm of mechanistic obscurity that will occupy future studies for years to come. © 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  4. Production of Dwarf Lettuce by Overexpressing a Pumpkin Gibberellin 20-Oxidase Gene

    PubMed Central

    Niki, Tomoya; Nishijima, Takaaki; Nakayama, Masayoshi; Hisamatsu, Tamotsu; Oyama-Okubo, Naomi; Yamazaki, Hiroko; Hedden, Peter; Lange, Theo; Mander, Lewis N.; Koshioka, Masaji

    2001-01-01

    We investigated the effect of overexpressing a pumpkin gibberellin (GA) 20-oxidase gene encoding an enzyme that forms predominantly biologically inactive products on GA biosynthesis and plant morphology in transgenic lettuce (Lactuca sativa cv Vanguard) plants. Lettuce was transformed with the pumpkin GA 20-oxidase gene downstream of a strong constitutive promoter cassette (El2–35S-Ω). The transgenic plants in which the pumpkin gene was detected by polymerase chain reaction were dwarfed in the T2 generation, whereas transformants with a normal growth phenotype did not contain the transgene. The result of Southern-blot analysis showed that the transgene was integrated as a single copy; the plants segregated three dwarfs to one normal in the T2 generation, indicating that the transgene was stable and dominant. The endogenous levels of GA1 and GA4 were reduced in the dwarfs, whereas large amounts of GA17 and GA25, which are inactive products of the pumpkin GA 20-oxidase, accumulated in these lines. These results indicate that a functional pumpkin GA 20-oxidase is expressed in the transgenic lettuce, resulting in a diversion of the normal pathway of GA biosynthesis to inactive products. Furthermore, this technique may be useful for controlling plant stature in other agricultural and horticultural species. PMID:11457947

  5. Ectopic expression of pumpkin gibberellin oxidases alters gibberellin biosynthesis and development of transgenic Arabidopsis plants.

    PubMed

    Radi, Abeer; Lange, Theo; Niki, Tomoya; Koshioka, Masaji; Lange, Maria João Pimenta

    2006-02-01

    Immature pumpkin (Cucurbita maxima) seeds contain gibberellin (GA) oxidases with unique catalytic properties resulting in GAs of unknown function for plant growth and development. Overexpression of pumpkin GA 7-oxidase (CmGA7ox) in Arabidopsis (Arabidopsis thaliana) resulted in seedlings with elongated roots, taller plants that flower earlier with only a little increase in bioactive GA4 levels compared to control plants. In the same way, overexpression of the pumpkin GA 3-oxidase1 (CmGA3ox1) resulted in a GA overdose phenotype with increased levels of endogenous GA4. This indicates that, in Arabidopsis, 7-oxidation and 3-oxidation are rate-limiting steps in GA plant hormone biosynthesis that control plant development. With an opposite effect, overexpression of pumpkin seed-specific GA 20-oxidase1 (CmGA20ox1) in Arabidopsis resulted in dwarfed plants that flower late with reduced levels of GA4 and increased levels of physiological inactive GA17 and GA25 and unexpected GA34 levels. Severe dwarfed plants were obtained by overexpression of the pumpkin GA 2-oxidase1 (CmGA2ox1) in Arabidopsis. This dramatic change in phenotype was accompanied by a considerable decrease in the levels of bioactive GA4 and an increase in the corresponding inactivation product GA34 in comparison to control plants. In this study, we demonstrate the potential of four pumpkin GA oxidase-encoding genes to modulate the GA plant hormone pool and alter plant stature and development.

  6. Identification and characterization of a novel flavin-containing spermine oxidase of mammalian cell origin.

    PubMed Central

    Vujcic, Slavoljub; Diegelman, Paula; Bacchi, Cyrus J; Kramer, Debora L; Porter, Carl W

    2002-01-01

    During polyamine catabolism, spermine and spermidine are first acetylated by spermidine/spermine N(1)-acetyltransferase (SSAT) and subsequently oxidized by polyamine oxidase (PAO) to produce spermidine and putrescine, respectively. In attempting to clone the PAO involved in this back-conversion pathway, we encountered an oxidase that preferentially cleaves spermine in the absence of prior acetylation by SSAT. A BLAST search using maize PAO sequences identified homologous mammalian cDNAs derived from human hepatoma and mouse mammary carcinoma: the encoded proteins differed by 20 amino acids. When either cDNA was transiently transfected into HEK-293 cells, intracellular spermine pools decreased by 75% while spermidine and N (1)-acetylspermidine pools increased, suggesting that spermine was selectively and directly oxidized by the enzyme. Substrate specificity using lysates of oxidase-transfected HEK-293 cells revealed that the newly identified oxidase strongly favoured spermine over N (1)-acetylspermine and that it failed to act on N (1)-acetylspermidine, spermidine or the preferred PAO substrate, N (1), N (12)-diacetylspermine. The PAO inhibitor, MDL-72,527, only partially blocked oxidation of spermine while a previously reported PAO substrate, N (1)-( n -octanesulphonyl)spermine, potently inhibited the reaction. Overall, the data indicate that the enzyme represents a novel mammalian oxidase which, on the basis of substrate specificity, we have designated spermine oxidase in order to distinguish it from the PAO involved in polyamine back-conversion. The identification of an enzyme capable of directly oxidizing spermine to spermidine has important implications for understanding polyamine homoeostasis and for interpreting metabolic and cellular responses to clinically relevant polyamine analogues and inhibitors. PMID:12141946

  7. Identification and biochemical characterization of polyamine oxidases in amphioxus: Implications for emergence of vertebrate-specific spermine and acetylpolyamine oxidases.

    PubMed

    Wang, Huihui; Liu, Baobao; Li, Hongyan; Zhang, Shicui

    2016-01-10

    Polyamine oxidases (PAOs) have been identified in a wide variety of animals, as well as in fungi and plant. Generally, plant PAOs oxidize spermine (Spm), spermidine (Spd) and their acetylated derivatives, N(1)-acetylspermine (N(1)-Aspm) and N(1)-acetylspermidine (N(1)-Aspd), while yeast PAOs oxidize Spm, N(1)-Aspm and N(1)-Aspd, but not Spd. By contrast, two different enzymes, namely spermine oxidase (SMO) and acetylpolyamine oxidase (APAO), specifically catalyze the oxidation of Spm and N(1)-Aspm/N(1)-Aspd, respectively. However, our knowledge on the biochemical and structural characterization of PAOs remains rather limited, and their evolutionary history is still enigmatic. In this study, two amphioxus (Branchiostoma japonicum) PAO genes, named Bjpao1 and Bjpao2, were cloned and characterized. Both Bjpao1 and Bjpao2 displayed distinct tissue-specific expression patterns. Notably, rBjPAO1 oxidized both spermine and spermidine, but not N(1)-acetylspermine, whereas rBjPAO2 oxidizes both spermidine and N(1)-acetylspermine, but not spermine. To understand structure-function relationship, the enzymatic activities of mutant BjPAOs that were generated by site-directed mutagenesis and expressed in E. coli were examined, The results indicate that the residues H64, K301 and T460 in rBjPAO1, and H69, K315 and T467 in rBjPAO2 were all involved in substrate binding and enzyme catalytic activity to some extent. Based on our results and those of others, a model depicting the divergent evolution and functional specialization of vertebrate SMO and APAO genes is proposed. Copyright © 2015 Elsevier B.V. All rights reserved.

  8. Molecular Characterization of the Fatty Alcohol Oxidation Pathway for Wax-Ester Mobilization in Germinated Jojoba Seeds1[W

    PubMed Central

    Rajangam, Alex S.; Gidda, Satinder K.; Craddock, Christian; Mullen, Robert T.; Dyer, John M.; Eastmond, Peter J.

    2013-01-01

    Jojoba (Simmondsia chinensis) is the only plant species known to use liquid wax esters (WEs) as a primary seed storage reserve. Upon germination, WE hydrolysis releases very-long-chain fatty alcohols, which must be oxidized to fatty acids by the sequential action of a fatty alcohol oxidase (FAO) and a fatty aldehyde dehydrogenase (FADH) before they can be β-oxidized. Here, we describe the cloning and characterization of genes for each of these two activities. Jojoba FAO and FADH are 52% and 68% identical to Arabidopsis (Arabidopsis thaliana) FAO3 and ALDH3H1, respectively. The genes are expressed most strongly in the cotyledons of jojoba seedlings following germination, but transcripts can also be detected in vegetative tissues. Proteomic analysis indicated that the FAO and FADH proteins can be detected on wax bodies, but they localized to the endoplasmic reticulum when they were expressed as amino-terminal green fluorescent protein fusions in tobacco (Nicotiana tabacum) leaves. Recombinant jojoba FAO and FADH proteins are active on very-long-chain fatty alcohol and fatty aldehyde substrates, respectively, and have biochemical properties consistent with those previously reported in jojoba cotyledons. Coexpression of jojoba FAO and FADH in Arabidopsis enhanced the in vivo rate of fatty alcohol oxidation more than 4-fold. Taken together, our data suggest that jojoba FAO and FADH constitute the very-long-chain fatty alcohol oxidation pathway that is likely to be necessary for efficient WE mobilization following seed germination. PMID:23166353

  9. Salicylic Acid Regulation of Respiration in Higher Plants: Alternative Oxidase Expression.

    PubMed Central

    Rhoads, DM; McIntosh, L

    1992-01-01

    Alternative respiratory pathway capacity increases during the development of the thermogenic appendix of a voodoo lily inflorescence. The levels of the alternative oxidase proteins increased dramatically between D-4 (4 days prior to the day of anthesis) and D-3 and continued to increase until the day of anthesis (D-day). The level of salicylic acid (SA) in the appendix is very low early on D-1, but increases to a high level in the evening of D-1. Thermogenesis occurs after a few hours of light on D-day. Therefore, the initial accumulation of the alternative oxidase proteins precedes the increase in SA by 3 days, indicating that other regulators may be involved. A 1.6-kb transcript encoding the alternative oxidase precursor protein accumulated to a high level in the appendix tissue by D-1. Application of SA to immature appendix tissue caused an increase in alternative pathway capacity and a dramatic accumulation of the alternative oxidase proteins and the 1.6-kb transcript. Time course experiments showed that the increase in capacity, protein levels, and transcript level corresponded precisely. The response to SA was blocked by cycloheximide or actinomycin D, indicating that de novo transcription and translation are required. However, nuclear, in vitro transcription assays indicated that the accumulation of the 1.6-kb transcript did not result from a simple increase in the rate of transcription of aox1. PMID:12297672

  10. The hemibiotrophic cacao pathogen Moniliophthora perniciosa depends on a mitochondrial alternative oxidase for biotrophic development

    USDA-ARS?s Scientific Manuscript database

    The mitochondrial alternative oxidase (AOX) is a non-energy conserving ubiquinol oxidase found in most fungal genomes studied to date. With the development of fungicides containing cytochrome-dependent respiratory chain (CRC) inhibitors, a strong interest in studying AOX functions in phytopathogenic...

  11. Combined effect of loss of the caa3 oxidase and Crp regulation drives Shewanella to thrive in redox-stratified environments.

    PubMed

    Zhou, Guangqi; Yin, Jianhua; Chen, Haijiang; Hua, Yijie; Sun, Linlin; Gao, Haichun

    2013-09-01

    Shewanella species are a group of facultative Gram-negative microorganisms with remarkable respiration abilities that allow the use of a diverse array of terminal electron acceptors (EA). Like most bacteria, S. oneidensis possesses multiple terminal oxidases, including two heme-copper oxidases (caa3- and cbb3-type) and a bd-type quinol oxidase. As aerobic respiration is energetically favored, mechanisms underlying the fact that these microorganisms thrive in redox-stratified environments remain vastly unexplored. In this work, we discovered that the cbb3-type oxidase is the predominant system for respiration of oxygen (O2), especially when O2 is abundant. Under microaerobic conditions, the bd-type quinol oxidase has a significant role in addition to the cbb3-type oxidase. In contrast, multiple lines of evidence suggest that under test conditions the caa3-type oxidase, an analog to the mitochondrial enzyme, has no physiological significance, likely because of its extremely low expression. In addition, expression of both cbb3- and bd-type oxidases is under direct control of Crp (cAMP receptor protein) but not the well-established redox regulator Fnr (fumarate nitrate regulator) of canonical systems typified in Escherichia coli. These data, collectively, suggest that adaptation of S. oneidensis to redox-stratified environments is likely due to functional loss of the caa3-type oxidase and switch of the regulatory system for respiration.

  12. Identification and statistical optimization of fermentation conditions for a newly isolated extracellular cholesterol oxidase-producing Streptomyces cavourensis strain NEAE-42.

    PubMed

    El-Naggar, Noura El-Ahmady; El-Shweihy, Nancy M; El-Ewasy, Sara M

    2016-09-20

    Due to broad range of clinical and industrial applications of cholesterol oxidase, isolation and screening of bacterial strains producing extracellular form of cholesterol oxidase is of great importance. One hundred and thirty actinomycete isolates were screened for their cholesterol oxidase activity. Among them, a potential culture, strain NEAE-42 is displayed the highest extracellular cholesterol oxidase activity. It was selected and identified as Streptomyces cavourensis strain NEAE-42. The optimization of different process parameters for cholesterol oxidase production by Streptomyces cavourensis strain NEAE-42 using Plackett-Burman experimental design and response surface methodology was carried out. Fifteen variables were screened using Plackett-Burman experimental design. Cholesterol, initial pH and (NH4)2SO4 were the most significant positive independent variables affecting cholesterol oxidase production. Central composite design was chosen to elucidate the optimal concentrations of the selected process variables on cholesterol oxidase production. It was found that, cholesterol oxidase production by Streptomyces cavourensis strain NEAE-42 after optimization process was 20.521U/mL which is higher than result obtained from the basal medium before screening process using Plackett-Burman (3.31 U/mL) with a fold of increase 6.19. The cholesterol oxidase level production obtained in this study (20.521U/mL) by the statistical method is higher than many of the reported values.

  13. IRON REGULATES XANTHINE OXIDASE ACTIVITY IN THE LUNG

    EPA Science Inventory

    The iron chelator deferoxamine has been reported to inhibit both xanthine oxidase (XO) and xanthine dehydrogenase activity, but the relationship of this effect to the availability of iron in the cellular and tissue environment remains unexplored. XO and total xanthine oxidoreduct...

  14. Molecular mechanisms of hypertension: role of Nox family NADPH oxidases.

    PubMed

    Sedeek, Mona; Hébert, Richard L; Kennedy, Chris R; Burns, Kevin D; Touyz, Rhian M

    2009-03-01

    Molecular mechanisms contributing to the pathoetiology of hypertension are complex, involving many interacting systems such as signaling through G protein-coupled receptors, the renin-angiotensin system, vascular inflammation and remodeling, vascular senescence and aging and developmental programming, as highlighted in the current issue of the journal. Common to these systems is NADPH oxidase-derived reactive oxygen species (ROS). This editorial highlights current concepts relating to the production of ROS in hypertension and focuses on the Nox family NADPH oxidases, major sources of free radicals in the cardiovascular and renal systems. ROS play a major role as intracellular signaling molecules to regulate normal biological cellular responses. In pathological conditions, loss of redox homeostasis contributes to vascular oxidative damage. Recent evidence indicates that specific enzymes, the Nox family of NADPH oxidases, have the sole function of generating ROS in a highly regulated fashion in physiological conditions, and that in disease states, hyperactivation of Noxes contributes to oxidative stress and consequent cardiovascular and renal injury. The Nox family comprises seven members, Nox1-Nox7. Nox1, Nox2 (gp91phox-containing NADPH oxidase), Nox4 and Nox5 have been identified in the cardiovascular-renal systems and have been implicated in the pathophysiology of cardiovascular and renal disease. Noxes, which are differentially regulated in hypertension, are major sources of cardiovascular and renal oxidative stress. This has evoked considerable interest because of the possibilities that therapies targeted against specific Nox isoforms to decrease ROS generation or to increase nitric oxide availability or both may be useful in minimizing vascular injury and renal dysfunction, and thereby prevent or regress target organ damage associated with hypertension.

  15. MAOA interacts with the ALDH2 gene in anxiety-depression alcohol dependence.

    PubMed

    Lee, Sheng-Yu; Hahn, Cheng-Yi; Lee, Jia-Fu; Huang, San-Yuan; Chen, Shiou-Lan; Kuo, Po-Hsiu; Lee, I Hui; Yeh, Tzung Lieh; Yang, Yen Kuang; Chen, Shih-Heng; Ko, Huei-Chen; Lu, Ru-Band

    2010-07-01

    Alcohol dependence is usually comorbid with anxiety disorder, depressive disorder, or both; this comorbidity may increase drinking behavior. We previously hypothesized that anxiety-depressive alcohol dependence (ANX/DEP ALC) was a genetically specific subtype of alcohol dependence. ANX/DEP ALC may be related to dopamine and serotonin, which are catalyzed by monoamine oxidase A (MAOA) and acetaldehyde dehydrogenase 2 (ALDH2). The aim of this study was to determine whether the interaction between the MAOA and the ALDH2 genes is associated with ANX/DEP ALC. We recruited 383 Han Chinese men in Taiwan: 143 ANX/DEP ALC and 240 healthy controls. The diagnosis of ANX/DEP ALC (alcohol dependence with a past or current history of anxiety, depressive disorder, or both) was made using DSM-IV criteria. Genotypes of ALDH2 and MAOA-uVNTR (variable number of tandem repeat located upstream) were determined using PCR-RFLP. The ALDH2, but not the MAOA-uVNTR, polymorphism was associated with ANX/DEP ALC. After stratifying the MAOA-uVNTR polymorphism, we found a stronger association between the ALDH2*1/*2 and *2/*2 genotypes and the controls in the MAOA-uVNTR 4-repeat subgroup. Logistic regression significantly associated the interaction between ALDH2 and MAOA variants with ANX/DEP ALC. We conclude that the MAOA and ALDH2 genes interact in ANX/DEP ALC. Although the MAOA gene alone is not associated with ANX/DEP ALC, we hypothesize that different variants of MAOA-uVNTR polymorphisms modify the protective effects of the ALDH2*2 allele on ANX/DEP ALC in Han Chinese in Taiwan.

  16. Alcoholism and alcohol drinking habits predicted from alcohol dehydrogenase genes.

    PubMed

    Tolstrup, Janne Schurmann; Nordestgaard, Børge Grønne; Rasmussen, Søren; Tybjaerg-Hansen, Anne; Grønbaek, Morten

    2008-06-01

    Alcohol drinking habits and alcoholism are partly genetically determined. Alcohol is degraded primarily by alcohol dehydrogenase (ADH) wherein genetic variation that affects the rate of alcohol degradation is found in ADH1B and ADH1C. It is biologically plausible that these variations may be associated with alcohol drinking habits and alcoholism. By genotyping 9080 white men and women from the general population, we found that men and women with ADH1B slow vs fast alcohol degradation drank more alcohol and had a higher risk of everyday drinking, heavy drinking, excessive drinking and of alcoholism. For example, the weekly alcohol intake was 9.8 drinks (95% confidence interval (CI): 9.1-11) among men with the ADH1B.1/1 genotype compared to 7.5 drinks (95% CI: 6.4-8.7) among men with the ADH1B.1/2 genotype, and the odds ratio (OR) for heavy drinking was 3.1 (95% CI: 1.7-5.7) among men with the ADH1B.1/1 genotype compared to men with the ADH1B.1/2 genotype. Furthermore, individuals with ADH1C slow vs fast alcohol degradation had a higher risk of heavy and excessive drinking. For example, the OR for heavy drinking was 1.4 (95% CI: 1.1-1.8) among men with the ADH1C.1/2 genotype and 1.4 (95% CI: 1.0-1.9) among men with the ADH1B.2/2 genotype, compared with men with the ADH1C.1/1 genotype. Results for ADH1B and ADH1C genotypes among men and women were similar. Finally, because slow ADH1B alcohol degradation is found in more than 90% of the white population compared to less than 10% of East Asians, the population attributable risk of heavy drinking and alcoholism by ADH1B.1/1 genotype was 67 and 62% among the white population compared with 9 and 24% among the East Asian population.

  17. Growth hormone and drug metabolism. Acute effects on microsomal mixed-function oxidase activities in rat liver.

    PubMed Central

    Wilson, J T; Spelsberg, T C

    1976-01-01

    Adult male rats were subjected either to sham operation or to hypophysectomy and adrenalectomy and maintained for a total of 10 days before treatment with growth hormone. Results of the early effects of growth hormone on the activities of the mixed-function oxidases in rat liver over a 96h period after growth-hormone treatment are presented. 2. Hypophysectomy and adrenalectomy result in decreased body and liver weight and decreased drug metabolism (mixed-function oxidases). Concentrations of electron-transport-system components are also decreased. 3. In the hypophysectomized/adrenalectomized rats, growth hormone decreases the activities of the liver mixed-function oxidases and the cytochrome P-450 and cytochrome c reductases, as well as decreasing the concentration of cytochrome P-450 compared with that of control rats. Similar but less dramatic results are obtained with sham-operated rats. 4. It is concluded that whereas growth hormone enhances liver growth, including induction of many enzyme activities, it results in a decrease in mixed-function oxidase activity. Apparently, mixed-function oxidase activity decreases in liver when growth (mitogenesis) increases. PMID:938458

  18. Highly ordered mesoporous carbons as electrode material for the construction of electrochemical dehydrogenase- and oxidase-based biosensors.

    PubMed

    Zhou, Ming; Shang, Li; Li, Bingling; Huang, Lijian; Dong, Shaojun

    2008-11-15

    In this work, the excellent catalytic activity of highly ordered mesoporous carbons (OMCs) to the electrooxidation of nicotinamide adenine dinucleotide (NADH) and hydrogen peroxide (H(2)O(2)) was described for the construction of electrochemical alcohol dehydrogenase (ADH) and glucose oxidase (GOD)-based biosensors. The high density of edge-plane-like defective sites and high specific surface area of OMCs could be responsible for the electrocatalytic behavior at OMCs modified glassy carbon electrode (OMCs/GE), which induced a substantial decrease in the overpotential of NADH and H(2)O(2) oxidation reaction compared to carbon nanotubes modified glassy carbon electrode (CNTs/GE). Such ability of OMCs permits effective low-potential amperometric biosensing of ethanol and glucose, respectively, at Nafion/ADH-OMCs/GE and Nafion/GOD-OMCs/GE. Especially, as an amperometric glucose biosensor, Nafion/GOD-OMCs/GE showed large determination range (500-15,000 micromoll(-1)), high sensitivity (0.053 nA micromol(-1)), fast (9+/-1s) and stable response (amperometric response retained 90% of the initial activity after 10h stirring of 2 mmoll(-1) glucose solution) to glucose as well as the effective discrimination to the possible interferences, which may make it to readily satisfy the need for the routine clinical diagnosis of diabetes. By comparing the electrochemical performance of OMCs with that of CNTs as electrode material for the construction of ADH- and GOD-biosensors in this work, we reveal that OMCs could be a favorable and promising carbon electrode material for constructing other electrochemical dehydrogenase- and oxidase-based biosensors, which may have wide potential applications in biocatalysis, bioelectronics and biofuel cells.

  19. Immunological and molecular comparison of polyphenol oxidase in Rosaceae fruit trees.

    PubMed

    Haruta, M; Murata, M; Kadokura, H; Homma, S

    1999-03-01

    An antibody raised against apple polyphenol oxidase (PPO) cross-reacted with PPOs from Japanese pear (Pyrus pyrifolia), pear (Pyrus communis), peach (Prunus persica), Chinese quince (Pseudocydonia sinensis) and Japanese loquat (Eriobotrya japonica). Core fragments (681 bp) of the corresponding PPO genes were amplified and characterized. The deduced protein sequences showed identities of 85.3 to 97.5%. Chlorogenic acid oxidase activity of these PPOs showed higher activities when assayed at pH 4 than at pH 6. These results indicate that PPOs in Rosaceae plants are structurally and enzymatically similar.

  20. A multicopper oxidase contributes to the copper tolerance of Brucella melitensis 16M.

    PubMed

    Wu, Tonglei; Wang, Shaohua; Wang, Zhen; Peng, Xiaowei; Lu, Yanli; Wu, Qingmin

    2015-06-01

    Copper is a potent antimicrobial agent. Multiple mechanisms of copper tolerance are utilized by some pathogenic bacteria. BMEII0580, which is significantly similar to the multicopper oxidase from Escherichia coli, was predicted to be the probable blue copper protein YacK precursor in Brucella melitensis 16M, and was designated as Brucella multicopper oxidase (BmcO). A bioinformatics analysis indicated that the typical motifs of multicopper oxidases are present in BmcO. BmcO, the expression of which was up-regulated by copper, could catalyze the oxidation of 2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), dimethoxyphenol (DMP) and para-phenylenediamine (pPD), which are widely used as substrates for multicopper oxidase. Additionally, BmcO exhibited ferroxidase activity, which indicated that it might play an important role in the Fe(2+) uptake of B. melitensis. Importantly, the mutant strain 16MΔbmcO was more sensitive to copper than the wild-type strain B. melitensis 16M as well as its complementation strain 16MΔbmcO(bmcO). The infection assays of cells showed that similar bacterial numbers of B. melitensis 16M, 16MΔbmcO and 16MΔbmcO(bmcO) strains were recovered from the infected macrophages. This result indicated that BmcO was not essential for B. melitensis intracellular growth. In conclusion, our results confirm that BmcO is a multicopper oxidase and contributes to the copper tolerance of B. melitensis 16M. © FEMS 2015. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.