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Sample records for nad nami ne

  1. NAMI

    Science.gov (United States)

    ... depressed. 'I’m just tired. I need a break,' went the script. But a break didn’t help." Read More Caregivers: When to ... to-Family NAMI Homefront NAMI In Our Own Voice NAMI Peer-to-Peer NAMI Parents & Teachers as ...

  2. "Me kohal on sinine taevas ..." = "Nebo sineje nad nami ..." : [luuletused] / Anne Lasikov ; tlk. Gennadi Krasnikov

    Index Scriptorium Estoniae

    Lasikov, Anne

    2003-01-01

    Sisu: "Me kohal on sinine taevas ..." = "Nebo sinee nad nami ..." ; "Siin valsirütme keerutasid jalad..." = "Kogda-to ritmõ valsa zdes kruzhili nogi ..." ; Sügisetuju = Osenneje nastroenije ; Ma tahan tagasi ; Kevad ; Suve algus ; Ood Narva-Jõesuule ; "V golovu lezjot vsjakaja drjan..." ; "Kogda vsbessilas avtorutshka..." ; "Ja ukrala tvoi vzgljad..." ; Zatshem mne radost tehh notshei? ; Vzgljad ; Tshernushka o sudbe ; "Zatshem võ, tshaiki, tak kritshite... " ; Nasledstvo ; Ja skisla.

  3. Steering Against Wind: A New Network of NamiRNAs and Enhancers

    Directory of Open Access Journals (Sweden)

    Ying Liang

    2017-10-01

    Full Text Available MicroRNAs (miRNAs are a class of endogenous non-coding RNAs with regulatory functions. Traditionally, miRNAs are thought to play a negative regulatory role in the cytoplasm by binding to the 3′UTR of target genes to degrade mRNA or inhibit translation. However, it remains a challenge to interpret the potential function of many miRNAs located in the nucleus. Recently, we reported a new type of miRNAs present in the nucleus, which can activate gene expression by binding to the enhancer, and named them nuclear activating miRNAs (NamiRNAs. The discovery of NamiRNAs showcases a complementary regulatory mechanism of miRNA, demonstrating their differential roles in the nucleus and cytoplasm. Here, we reviewed miRNAs in nucleus to better understand the function of NamiRNAs in their interactions with the enhancers. Accordingly, we propose a NamiRNA–enhancer–target gene activation network model to better understand the crosstalk between NamiRNAs and enhancers in regulating gene transcription. Moreover, we hypothesize that NamiRNAs may be involved in cell identity or cell fate determination during development, although further study is needed to elucidate the underlying mechanisms in detail. Keywords: Nuclear activating miRNAs, Tissue-specific enhancers, Transcriptional gene activation, Cell identity, Cell fate

  4. Pharmaceutical development of the novel metal-based anticancer agents NAMI-A and AP 5280

    NARCIS (Netherlands)

    Bouma, M. (Marjan)

    2002-01-01

    The pharmaceutical development of the two novel metal-based anticancer agents NAMI-A and AP 5280 is described in this thesis, starting with the development of analytical methods for the quality control of drug substance and final product, via the formulation process leading to a stable, intravenous

  5. Inhibitory effects of NAMI-A-like ruthenium complexes on prion neuropeptide fibril formation.

    Science.gov (United States)

    Wang, Xuesong; Zhu, Dengsen; Zhao, Cong; He, Lei; Du, Weihong

    2015-05-01

    Prion diseases are a group of infectious and fatal neurodegenerative disorders caused by the conformational conversion of a cellular prion protein (PrP) into its abnormal isoform PrP(Sc). PrP106-126 resembles PrP(Sc) in terms of physicochemical and biological characteristics and is used as a common model for the treatment of prion diseases. Inhibitory effects on fibril formation and neurotoxicity of the prion neuropeptide PrP106-126 have been investigated using metal complexes as potential inhibitors. Nevertheless, the binding mechanism between metal complexes and the peptide remains unclear. The present study is focused on the interaction of PrP106-126 with NAMI-A and NAMI-A-like ruthenium complexes, including KP418, KP1019, and KP1019-2. Results demonstrated that these ruthenium complexes could bind to PrP106-126 in a distinctive binding mode through electrostatic and hydrophobic interactions. NAMI-A-like ruthenium complexes can also effectively inhibit the aggregation and fibril formation of PrP106-126. The complex KP1019 demonstrated the optimal inhibitory ability upon peptide aggregation, and cytotoxicity because of its large aromatic ligand contribution. The studied complexes could also regulate the copper redox chemistry of PrP106-126 and effectually inhibit the formation of reactive oxygen species. Given these findings, ruthenium complexes with relatively low cellular toxicity may be used to develop potential pharmaceutical products against prion diseases.

  6. Whole-Body Counter(WBC and food radiocesium contamination surveys in Namie, Fukushima Prefecture.

    Directory of Open Access Journals (Sweden)

    Yoichiro Hosokawa

    Full Text Available This study examined the internal Cs exposure of residents and the Cs present in food products produced in Namie. Whole-body counter (WBC was used for the measurement of internal exposure per each whole body of examinees.The food products which appeared to be used for consumption, were brought by residents and commercially available food items were excluded. Most of them were wild plants or food items produced by residents. Four years of data from April 2012 to March 2013 (fiscal 2012 and April 2015 to March 2016 (Fiscal 2015 were analyzed and studied.The average radioactivity measured by WBC was approximately 5 Bq for Cs-134, and 20 Bq for Cs-137 and the average committed effective dose was approximately 1 μSv. The average for the residents with detectable radioactivity was 25 μSv, and the human health effects are considered to be extremely low risk. However, the radioactivity of the affected individuals showed a higher value than the theoretical attenuation rate. The majority (83.2% of individuals exhibiting radioactivity were over 50 years old. The number of food products brought in for detection decreased as the study period progressed, but the number of food products with radioactivity had increased. While the items with a higher detection rate of radioactivity included fruits such as citron and persimmon, shiitake mushrooms exhibited the highest radioactivity. Moreover, the radioactivity of seven items in these 10 items decreased from fiscal 2012 to fiscal 2015. Mushrooms had high radioactivity and were produced over a wide area.We suggest that the elderly try to enjoy life and eat wild plants in moderation while inspecting food products. Therefore, we will continue to work in raising awareness of radiation and its potential presence in food products and thus the continuing necessity of monitoring radioactivity in food in the future.

  7. Analysis of chromosome translocation in the residents of Namie Town after the accident of Fukushima Daiichi Nuclear Power Plant

    International Nuclear Information System (INIS)

    Yoshida, Mitsuaki A.; Nakata, Akifumi; Miura, Tomisato; Nishimura, Miya; Takamagi, Shizuka; Kasai, Kosuke; Konno, Norio; Yoshida, Ryoko; Sekine, Shunji

    2014-01-01

    The dose estimation by behavior survey of the residents carried out by Fukushima Prefecture after the accident reported that there are no residents who were exposed by over 1 mSv radiation. However, a lot of the parents are worrying about the health condition of their children in future. Our Hirosaki University accepted the request of the local government of this Namie-Town in Fukushima which wants to know whether children were exposed by radiological substances or not and started the inspection about the contamination and exposure level and dose estimation at an accident using chromosomal translocation analysis for 855 out of 3700 children whose age was under 18 years old at the time of accident. In order to estimate radiation dose using chromosome aberration in the accidents, there are four kinds of cytogenetic method; dicentric assay, a translocation assay, the PCC-ring assay and micronucleus test. A dicentric assay is used for the dose estimation in acute and external exposure cases, the chromosomal translocation method for dose assessment in chronic and old exposure and the PCC method for high dose exposure, respectively. In the case of the residents in Namie-Town, since about one year and ten months had already passed after the accident when implementation of this inspection was determined, the chromosomal translocation method was applied for the dose estimation of the initial exposure level. The main purpose of this translocation analysis using their own cells is to take away affairs of the residents including parents and children and also to reduce the uneasiness which is not wiped away by the health check due to a behavioral survey. In this inspection, after the contents and process of this analysis were explained in the Tsushima, Namie-Town temporarily constructed clinic, 3∼4 ml of whole 5 blood were taken from each children whose parents agreed with this analysis. The lymphocytic cells are isolated from the whole blood using CPT (Cell Preparation Tube

  8. Boosting NAD to spare hearing.

    Science.gov (United States)

    Brenner, Charles

    2014-12-02

    Ex vivo experiments have strangely shown that inhibition or stimulation of NAD metabolism can be neuroprotective. In this issue of Cell Metabolism, Brown et al. (2014) demonstrate that cochlear NAD is diminished by deafening noise but protected by nicotinamide riboside or WldS mutation. Hearing protection by nicotinamide riboside depends on Sirt3. Copyright © 2014 Elsevier Inc. All rights reserved.

  9. "Zolotogo dozhdja" nad "stalinskim ampirom" ne budet / Nadezhda Ivantsova

    Index Scriptorium Estoniae

    Ivantsova, Nadezhda

    2003-01-01

    2002. a. kuulutati kinoteater "Rodina" ja kultuurimaja Sillamäel arhitektuurimälestusmärkideks kui stalinliku ampiiri näited. Nende renoveerimise ja korrashoiuga seotud probleemidest. Kommentaar Vladimir Shurminilt

  10. VT NAD83 USGS Quadrangle Boundaries - polygons

    Data.gov (United States)

    Vermont Center for Geographic Information — (Link to Metadata) Generated from exact latitude-longitude coordinates and projected from Geographic coordinates (Lat/Long) NAD83 into State Plane Meters NAD83. The...

  11. Gennadi Meleshko predstavil film "Poljot nad vremenjem"

    Index Scriptorium Estoniae

    2007-01-01

    Teleajakirjanike Gennadi ja Jelena Meleshko dokumentaalfilmi "Lend aja kohal" ("Poljot nad vremenem") esitleti 21. novembril Eesti Rahvusraamatukogus. Tervitusega esines ka riigikogu liigr Peeter Tulviste

  12. Vitamins and aging: pathways to NAD+ synthesis.

    Science.gov (United States)

    Denu, John M

    2007-05-04

    Recent genetic evidence reveals additional salvage pathways for NAD(+) synthesis. In this issue, Belenky et al. (2007) report that nicotinamide riboside, a new NAD(+) precursor, regulates Sir2 deacetylase activity and life span in yeast. The ability of nicotinamide riboside to enhance life span does not depend on calorie restriction.

  13. Why NAD(+) Declines during Aging: It's Destroyed.

    Science.gov (United States)

    Schultz, Michael B; Sinclair, David A

    2016-06-14

    NAD(+) is required not only for life but for a long life. In this issue, Camacho-Pereira et al. (2016) implicate CD38 in the decline of NAD(+) during aging, with implications for combating age-related diseases. Copyright © 2016 Elsevier Inc. All rights reserved.

  14. NAD+ in Aging

    DEFF Research Database (Denmark)

    Fang, Evandro F.; Lautrup, Sofie; Hou, Yujun

    2017-01-01

    neuronal function. NAD+ depletion is detected in major neurodegenerative diseases, such as Alzheimer's and Parkinson's diseases, cardiovascular disease and muscle atrophy. Emerging evidence suggests that NAD+ decrements occur in various tissues during aging, and that physiological and pharmacological...... interventions bolstering cellular NAD+ levels might retard aspects of aging and forestall some age-related diseases. Here, we discuss aspects of NAD+ biosynthesis, together with putative mechanisms of NAD+ action against aging, including recent preclinical and clinical trials. Recent discoveries have...... DNA repair and mitochondrial maintenance. Mitochondrial autophagy (mitophagy) has a major role in clearance of damaged and/or dysfunctional mitochondria, and compromised mitophagy has been linked to metabolic disorders, neurodegeneration [including Alzheimer's disease (AD) and Parkinson's disease (PD...

  15. VT NAD83 Orthophoto Boundaries - corner points

    Data.gov (United States)

    Vermont Center for Geographic Information — (Link to Metadata) RF 5000 NAD83 orthophoto edge lines (4000 x 4000 meter grid cells) were generated automatically from the known corner locations (generated by Gary...

  16. VT NAD83 Orthophoto Boundaries - polygons

    Data.gov (United States)

    Vermont Center for Geographic Information — (Link to Metadata) RF 5000 NAD83 orthophoto edge lines (4000 x 4000 meter grid cells) were generated automatically from the known corner locations (generated by Gary...

  17. United States Stateplane Zones - NAD27

    Data.gov (United States)

    Earth Data Analysis Center, University of New Mexico — U.S. State Plane Zones (NAD 1927) represents the State Plane Coordinate System (SPCS) Zones for the 1927 North American Datum within United States.

  18. United States Stateplane Zones - NAD83

    Data.gov (United States)

    Earth Data Analysis Center, University of New Mexico — U.S. State Plane Zones (NAD 1983) represents the State Plane Coordinate System (SPCS) Zones for the 1983 North American Datum within United States.

  19. Glutamine versus ammonia utilization in the NAD synthetase family.

    Directory of Open Access Journals (Sweden)

    Jessica De Ingeniis

    Full Text Available NAD is a ubiquitous and essential metabolic redox cofactor which also functions as a substrate in certain regulatory pathways. The last step of NAD synthesis is the ATP-dependent amidation of deamido-NAD by NAD synthetase (NADS. Members of the NADS family are present in nearly all species across the three kingdoms of Life. In eukaryotic NADS, the core synthetase domain is fused with a nitrilase-like glutaminase domain supplying ammonia for the reaction. This two-domain NADS arrangement enabling the utilization of glutamine as nitrogen donor is also present in various bacterial lineages. However, many other bacterial members of NADS family do not contain a glutaminase domain, and they can utilize only ammonia (but not glutamine in vitro. A single-domain NADS is also characteristic for nearly all Archaea, and its dependence on ammonia was demonstrated here for the representative enzyme from Methanocaldococcus jannaschi. However, a question about the actual in vivo nitrogen donor for single-domain members of the NADS family remained open: Is it glutamine hydrolyzed by a committed (but yet unknown glutaminase subunit, as in most ATP-dependent amidotransferases, or free ammonia as in glutamine synthetase? Here we addressed this dilemma by combining evolutionary analysis of the NADS family with experimental characterization of two representative bacterial systems: a two-subunit NADS from Thermus thermophilus and a single-domain NADS from Salmonella typhimurium providing evidence that ammonia (and not glutamine is the physiological substrate of a typical single-domain NADS. The latter represents the most likely ancestral form of NADS. The ability to utilize glutamine appears to have evolved via recruitment of a glutaminase subunit followed by domain fusion in an early branch of Bacteria. Further evolution of the NADS family included lineage-specific loss of one of the two alternative forms and horizontal gene transfer events. Lastly, we identified NADS

  20. Assimilation of NAD(+) precursors in Candida glabrata.

    Science.gov (United States)

    Ma, Biao; Pan, Shih-Jung; Zupancic, Margaret L; Cormack, Brendan P

    2007-10-01

    The yeast pathogen Candida glabrata is a nicotinamide adenine dinucleotide (NAD(+)) auxotroph and its growth depends on the environmental supply of vitamin precursors of NAD(+). C. glabrata salvage pathways defined in this article allow NAD(+) to be synthesized from three compounds - nicotinic acid (NA), nicotinamide (NAM) and nicotinamide riboside (NR). NA is salvaged through a functional Preiss-Handler pathway. NAM is first converted to NA by nicotinamidase and then salvaged by the Preiss-Handler pathway. Salvage of NR in C. glabrata occurs via two routes. The first, in which NR is phosphorylated by the NR kinase Nrk1, is independent of the Preiss-Handler pathway. The second is a novel pathway in which NR is degraded by the nucleosidases Pnp1 and Urh1, with a minor role for Meu1, and ultimately converted to NAD(+) via the nicotinamidase Pnc1 and the Preiss-Handler pathway. Using C. glabrata mutants whose growth depends exclusively on the external NA or NR supply, we also show that C. glabrata utilizes NR and to a lesser extent NA as NAD(+) sources during disseminated infection.

  1. VT NAD83 USGS Quadrangle Boundaries - corner points

    Data.gov (United States)

    Vermont Center for Geographic Information — (Link to Metadata) Generated from exact latitude-longitude coordinates and projected from Geographic coordinates (Lat/Long) NAD83 into State Plane Meters NAD83. The...

  2. NAD Deficiency, Congenital Malformations, and Niacin Supplementation.

    Science.gov (United States)

    Shi, Hongjun; Enriquez, Annabelle; Rapadas, Melissa; Martin, Ella M M A; Wang, Roni; Moreau, Julie; Lim, Chai K; Szot, Justin O; Ip, Eddie; Hughes, James N; Sugimoto, Kotaro; Humphreys, David T; McInerney-Leo, Aideen M; Leo, Paul J; Maghzal, Ghassan J; Halliday, Jake; Smith, Janine; Colley, Alison; Mark, Paul R; Collins, Felicity; Sillence, David O; Winlaw, David S; Ho, Joshua W K; Guillemin, Gilles J; Brown, Matthew A; Kikuchi, Kazu; Thomas, Paul Q; Stocker, Roland; Giannoulatou, Eleni; Chapman, Gavin; Duncan, Emma L; Sparrow, Duncan B; Dunwoodie, Sally L

    2017-08-10

    Congenital malformations can be manifested as combinations of phenotypes that co-occur more often than expected by chance. In many such cases, it has proved difficult to identify a genetic cause. We sought the genetic cause of cardiac, vertebral, and renal defects, among others, in unrelated patients. We used genomic sequencing to identify potentially pathogenic gene variants in families in which a person had multiple congenital malformations. We tested the function of the variant by using assays of in vitro enzyme activity and by quantifying metabolites in patient plasma. We engineered mouse models with similar variants using the CRISPR (clustered regularly interspaced short palindromic repeats)-Cas9 system. Variants were identified in two genes that encode enzymes of the kynurenine pathway, 3-hydroxyanthranilic acid 3,4-dioxygenase (HAAO) and kynureninase (KYNU). Three patients carried homozygous variants predicting loss-of-function changes in the HAAO or KYNU proteins (HAAO p.D162*, HAAO p.W186*, or KYNU p.V57Efs*21). Another patient carried heterozygous KYNU variants (p.Y156* and p.F349Kfs*4). The mutant enzymes had greatly reduced activity in vitro. Nicotinamide adenine dinucleotide (NAD) is synthesized de novo from tryptophan through the kynurenine pathway. The patients had reduced levels of circulating NAD. Defects similar to those in the patients developed in the embryos of Haao-null or Kynu-null mice owing to NAD deficiency. In null mice, the prevention of NAD deficiency during gestation averted defects. Disruption of NAD synthesis caused a deficiency of NAD and congenital malformations in humans and mice. Niacin supplementation during gestation prevented the malformations in mice. (Funded by the National Health and Medical Research Council of Australia and others.).

  3. NAD+ biosynthesis, aging, and disease [version 1; referees: 2 approved

    OpenAIRE

    Sean Johnson; Shin–ichiro Imai

    2018-01-01

    Nicotinamide adenine dinucleotide (NAD+) biosynthesis and its regulation have recently been attracting markedly increasing interest. Aging is marked by a systemic decrease in NAD+ across multiple tissues. The dysfunction of NAD+ biosynthesis plays a critical role in the pathophysiologies of multiple diseases, including age-associated metabolic disorders, neurodegenerative diseases, and mental disorders. As downstream effectors, NAD+-dependent enzymes, such as sirtuins, are involved in the pro...

  4. Loss of NAD(H from swollen yeast mitochondria

    Directory of Open Access Journals (Sweden)

    Pfeiffer Douglas R

    2006-01-01

    Full Text Available Abstract Background The mitochondrial electron transport chain oxidizes matrix space NADH as part of the process of oxidative phosphorylation. Mitochondria contain shuttles for the transport of cytoplasmic NADH reducing equivalents into the mitochondrial matrix. Therefore for a long time it was believed that NAD(H itself was not transported into mitochondria. However evidence has been obtained for the transport of NAD(H into and out of plant and mammalian mitochondria. Since Saccharomyces cerevisiae mitochondria can directly oxidize cytoplasmic NADH, it remained questionable if mitochondrial NAD(H transport occurs in this organism. Results NAD(H was lost more extensively from the matrix space of swollen than normal, condensed isolated yeast mitochondria from Saccharomyces cerevisiae. The loss of NAD(H in swollen organelles caused a greatly decreased respiratory rate when ethanol or other matrix space NAD-linked substrates were oxidized. Adding NAD back to the medium, even in the presence of a membrane-impermeant NADH dehydrogenase inhibitor, restored the respiratory rate of swollen mitochondria oxidizing ethanol, suggesting that NAD is transported into the matrix space. NAD addition did not restore the decreased respiratory rate of swollen mitochondria oxidizing the combination of malate, glutamate, and pyruvate. Therefore the loss of matrix space metabolites is not entirely specific for NAD(H. However, during NAD(H loss the mitochondrial levels of most other nucleotides were maintained. Either hypotonic swelling or colloid-osmotic swelling due to opening of the yeast mitochondrial unspecific channel (YMUC in a mannitol medium resulted in decreased NAD-linked respiration. However, the loss of NAD(H from the matrix space was not mediated by the YMUC, because YMUC inhibitors did not prevent decreased NAD-linked respiration during swelling and YMUC opening without swelling did not cause decreased NAD-linked respiration. Conclusion Loss of

  5. The evolutionary portrait of metazoan NAD salvage.

    Science.gov (United States)

    Carneiro, João; Duarte-Pereira, Sara; Azevedo, Luísa; Castro, L Filipe C; Aguiar, Paulo; Moreira, Irina S; Amorim, António; Silva, Raquel M

    2013-01-01

    Nicotinamide Adenine Dinucleotide (NAD) levels are essential for cellular homeostasis and survival. Main sources of intracellular NAD are the salvage pathways from nicotinamide, where Nicotinamide phosphoribosyltransferases (NAMPTs) and Nicotinamidases (PNCs) have a key role. NAMPTs and PNCs are important in aging, infection and disease conditions such as diabetes and cancer. These enzymes have been considered redundant since either one or the other exists in each individual genome. The co-occurrence of NAMPT and PNC was only recently detected in invertebrates though no structural or functional characterization exists for them. Here, using expression and evolutionary analysis combined with homology modeling and protein-ligand docking, we show that both genes are expressed simultaneously in key species of major invertebrate branches and emphasize sequence and structural conservation patterns in metazoan NAMPT and PNC homologues. The results anticipate that NAMPTs and PNCs are simultaneously active, raising the possibility that NAD salvage pathways are not redundant as both are maintained to fulfill the requirement for NAD production in some species.

  6. The evolutionary portrait of metazoan NAD salvage.

    Directory of Open Access Journals (Sweden)

    João Carneiro

    Full Text Available Nicotinamide Adenine Dinucleotide (NAD levels are essential for cellular homeostasis and survival. Main sources of intracellular NAD are the salvage pathways from nicotinamide, where Nicotinamide phosphoribosyltransferases (NAMPTs and Nicotinamidases (PNCs have a key role. NAMPTs and PNCs are important in aging, infection and disease conditions such as diabetes and cancer. These enzymes have been considered redundant since either one or the other exists in each individual genome. The co-occurrence of NAMPT and PNC was only recently detected in invertebrates though no structural or functional characterization exists for them. Here, using expression and evolutionary analysis combined with homology modeling and protein-ligand docking, we show that both genes are expressed simultaneously in key species of major invertebrate branches and emphasize sequence and structural conservation patterns in metazoan NAMPT and PNC homologues. The results anticipate that NAMPTs and PNCs are simultaneously active, raising the possibility that NAD salvage pathways are not redundant as both are maintained to fulfill the requirement for NAD production in some species.

  7. NAD+ in Aging: Molecular Mechanisms and Translational Implications.

    Science.gov (United States)

    Fang, Evandro F; Lautrup, Sofie; Hou, Yujun; Demarest, Tyler G; Croteau, Deborah L; Mattson, Mark P; Bohr, Vilhelm A

    2017-10-01

    The coenzyme NAD + is critical in cellular bioenergetics and adaptive stress responses. Its depletion has emerged as a fundamental feature of aging that may predispose to a wide range of chronic diseases. Maintenance of NAD + levels is important for cells with high energy demands and for proficient neuronal function. NAD + depletion is detected in major neurodegenerative diseases, such as Alzheimer's and Parkinson's diseases, cardiovascular disease and muscle atrophy. Emerging evidence suggests that NAD + decrements occur in various tissues during aging, and that physiological and pharmacological interventions bolstering cellular NAD + levels might retard aspects of aging and forestall some age-related diseases. Here, we discuss aspects of NAD + biosynthesis, together with putative mechanisms of NAD + action against aging, including recent preclinical and clinical trials. Published by Elsevier Ltd.

  8. NAD Acts as an Integral Regulator of Multiple Defense Layers.

    Science.gov (United States)

    Pétriacq, Pierre; Ton, Jurriaan; Patrit, Oriane; Tcherkez, Guillaume; Gakière, Bertrand

    2016-11-01

    Pyridine nucleotides, such as NAD, are crucial redox carriers and have emerged as important signaling molecules in stress responses. Previously, we have demonstrated in Arabidopsis (Arabidopsis thaliana) that the inducible NAD-overproducing nadC lines are more resistant to an avirulent strain of Pseudomonas syringae pv tomato (Pst-AvrRpm1), which was associated with salicylic acid-dependent defense. Here, we have further characterized the NAD-dependent immune response in Arabidopsis. Quinolinate-induced stimulation of intracellular NAD in transgenic nadC plants enhanced resistance against a diverse range of (a)virulent pathogens, including Pst-AvrRpt2, Dickeya dadantii, and Botrytis cinerea Characterization of the redox status demonstrated that elevated NAD levels induce reactive oxygen species (ROS) production and the expression of redox marker genes of the cytosol and mitochondrion. Using pharmacological and reverse genetics approaches, we show that NAD-induced ROS production functions independently of NADPH oxidase activity and light metabolism but depends on mitochondrial respiration, which was increased at higher NAD. We further demonstrate that NAD primes pathogen-induced callose deposition and cell death. Mass spectrometry analysis reveals that NAD simultaneously induces different defense hormones and that the NAD-induced metabolic profiles are similar to those of defense-expressing plants after treatment with pathogen-associated molecular patterns. We thus conclude that NAD triggers metabolic profiles rather similar to that of pathogen-associated molecular patterns and discuss how signaling cross talk between defense hormones, ROS, and NAD explains the observed resistance to pathogens. © 2016 American Society of Plant Biologists. All Rights Reserved.

  9. Photoionization of Ne8+

    Science.gov (United States)

    Pindzola, M. S.; Abdel-Naby, Sh. A.; Robicheaux, F.; Colgan, J.

    2014-05-01

    Single and double photoionization cross sections for Ne8+ are calculated using a non-perturbative fully relativistic time-dependent close-coupling method. A Bessel function expansion is used to include both dipole and quadrupole effects in the radiation field interaction and the repulsive interaction between electrons includes both the Coulomb and Gaunt interactions. The fully correlated ground state of Ne8+ is obtained by solving a time-independent inhomogeneous set of close-coupled equations. Propagation of the time-dependent close-coupled equations yields single and double photoionization cross sections for Ne8+ at energies easily accessible at advanced free electron laser facilities. This work was supported in part by grants from NSF and US DoE. Computational work was carried out at NERSC in Oakland, California, NICS in Knoxville, Tennessee, and OLCF in Oak Ridge, Tennessee.

  10. NAD+metabolism: Bioenergetics, signaling and manipulation for therapy.

    Science.gov (United States)

    Yang, Yue; Sauve, Anthony A

    2016-12-01

    We survey the historical development of scientific knowledge surrounding Vitamin B3, and describe the active metabolite forms of Vitamin B3, the pyridine dinucleotides NAD + and NADP + which are essential to cellular processes of energy metabolism, cell protection and biosynthesis. The study of NAD + has become reinvigorated by new understandings that dynamics within NAD + metabolism trigger major signaling processes coupled to effectors (sirtuins, PARPs, and CD38) that reprogram cellular metabolism using NAD + as an effector substrate. Cellular adaptations include stimulation of mitochondrial biogenesis, a process fundamental to adjusting cellular and tissue physiology to reduced nutrient availability and/or increased energy demand. Several mammalian metabolic pathways converge to NAD + , including tryptophan-derived de novo pathways, nicotinamide salvage pathways, nicotinic acid salvage and nucleoside salvage pathways incorporating nicotinamide riboside and nicotinic acid riboside. Key discoveries highlight a therapeutic potential for targeting NAD + biosynthetic pathways for treatment of human diseases. A recent emergence of understanding that NAD + homeostasis is vulnerable to aging and disease processes has stimulated testing to determine if replenishment or augmentation of cellular or tissue NAD + can have ameliorative effects on aging or disease phenotypes. This experimental approach has provided several proofs of concept successes demonstrating that replenishment or augmentation of NAD + concentrations can provide ameliorative or curative benefits. Thus NAD + metabolic pathways can provide key biomarkers and parameters for assessing and modulating organism health. Copyright © 2016. Published by Elsevier B.V.

  11. Nrt1 and Tna1-independent export of NAD+ precursor vitamins promotes NAD+ homeostasis and allows engineering of vitamin production.

    Directory of Open Access Journals (Sweden)

    Peter Belenky

    2011-05-01

    Full Text Available NAD(+ is both a co-enzyme for hydride transfer enzymes and a substrate of sirtuins and other NAD(+ consuming enzymes. NAD(+ biosynthesis is required for two different regimens that extend lifespan in yeast. NAD(+ is synthesized from tryptophan and the three vitamin precursors of NAD(+: nicotinic acid, nicotinamide and nicotinamide riboside. Supplementation of yeast cells with NAD(+ precursors increases intracellular NAD(+ levels and extends replicative lifespan. Here we show that both nicotinamide riboside and nicotinic acid are not only vitamins but are also exported metabolites. We found that the deletion of the nicotinamide riboside transporter, Nrt1, leads to increased export of nicotinamide riboside. This discovery was exploited to engineer a strain to produce high levels of extracellular nicotinamide riboside, which was recovered in purified form. We further demonstrate that extracellular nicotinamide is readily converted to extracellular nicotinic acid in a manner that requires intracellular nicotinamidase activity. Like nicotinamide riboside, export of nicotinic acid is elevated by the deletion of the nicotinic acid transporter, Tna1. The data indicate that NAD(+ metabolism has a critical extracellular element in the yeast system and suggest that cells regulate intracellular NAD(+ metabolism by balancing import and export of NAD(+ precursor vitamins.

  12. Coupling of NAD+ biosynthesis and nicotinamide ribosyl transport: characterization of NadR ribonucleotide kinase mutants of Haemophilus influenzae.

    Science.gov (United States)

    Merdanovic, Melisa; Sauer, Elizabeta; Reidl, Joachim

    2005-07-01

    Previously, we characterized a pathway necessary for the processing of NAD+ and for uptake of nicotinamide riboside (NR) in Haemophilus influenzae. Here we report on the role of NadR, which is essential for NAD+ utilization in this organism. Different NadR variants with a deleted ribonucleotide kinase domain or with a single amino acid change were characterized in vitro and in vivo with respect to cell viability, ribonucleotide kinase activity, and NR transport. The ribonucleotide kinase mutants were viable only in a nadV+ (nicotinamide phosphoribosyltransferase) background, indicating that the ribonucleotide kinase domain is essential for cell viability in H. influenzae. Mutations located in the Walker A and B motifs and the LID region resulted in deficiencies in both NR phosphorylation and NR uptake. The ribonucleotide kinase function of NadR was found to be feedback controlled by NAD+ under in vitro conditions and by NAD+ utilization in vivo. Taken together, our data demonstrate that the NR phosphorylation step is essential for both NR uptake across the inner membrane and NAD+ synthesis and is also involved in controlling the NAD+ biosynthesis rate.

  13. Nrt1 and Tna1-independent export of NAD+ precursor vitamins promotes NAD+ homeostasis and allows engineering of vitamin production.

    Science.gov (United States)

    Belenky, Peter; Stebbins, Rebecca; Bogan, Katrina L; Evans, Charles R; Brenner, Charles

    2011-05-11

    NAD(+) is both a co-enzyme for hydride transfer enzymes and a substrate of sirtuins and other NAD(+) consuming enzymes. NAD(+) biosynthesis is required for two different regimens that extend lifespan in yeast. NAD(+) is synthesized from tryptophan and the three vitamin precursors of NAD(+): nicotinic acid, nicotinamide and nicotinamide riboside. Supplementation of yeast cells with NAD(+) precursors increases intracellular NAD(+) levels and extends replicative lifespan. Here we show that both nicotinamide riboside and nicotinic acid are not only vitamins but are also exported metabolites. We found that the deletion of the nicotinamide riboside transporter, Nrt1, leads to increased export of nicotinamide riboside. This discovery was exploited to engineer a strain to produce high levels of extracellular nicotinamide riboside, which was recovered in purified form. We further demonstrate that extracellular nicotinamide is readily converted to extracellular nicotinic acid in a manner that requires intracellular nicotinamidase activity. Like nicotinamide riboside, export of nicotinic acid is elevated by the deletion of the nicotinic acid transporter, Tna1. The data indicate that NAD(+) metabolism has a critical extracellular element in the yeast system and suggest that cells regulate intracellular NAD(+) metabolism by balancing import and export of NAD(+) precursor vitamins.

  14. NAD+ biosynthesis, aging, and disease [version 1; referees: 2 approved

    Directory of Open Access Journals (Sweden)

    Sean Johnson

    2018-02-01

    Full Text Available Nicotinamide adenine dinucleotide (NAD+ biosynthesis and its regulation have recently been attracting markedly increasing interest. Aging is marked by a systemic decrease in NAD+ across multiple tissues. The dysfunction of NAD+ biosynthesis plays a critical role in the pathophysiologies of multiple diseases, including age-associated metabolic disorders, neurodegenerative diseases, and mental disorders. As downstream effectors, NAD+-dependent enzymes, such as sirtuins, are involved in the progression of such disorders. These recent studies implicate NAD+ biosynthesis as a potential target for preventing and treating age-associated diseases. Indeed, new studies have demonstrated the therapeutic potential of supplementing NAD+ intermediates, such as nicotinamide mononucleotide and nicotinamide riboside, providing a proof of concept for the development of an effective anti-aging intervention.

  15. Nad avantjuristom godõ ne vlastnõ / Boris Tuch

    Index Scriptorium Estoniae

    Tuch, Boris, 1946-

    2008-01-01

    Steven Spielbergi neljas Indiana Jones'i film Harrison Fordiga nimiosas "Indiana Jones ja kristallpealuu kuningriik" ("Indiana Jones and the Kingdom of the Crystal Skull") : Ameerika Ühendriigid 2008

  16. Why NAD+ Declines during Aging: It’s Destroyed

    Science.gov (United States)

    Schultz, Michael B.; Sinclair, David A.

    2016-01-01

    NAD+ is required not only for life but for a long life. In this issue, Camacho-Pereira et al. (2016) implicate CD38 in the decline of NAD+ during aging, with implications for combating age-related diseases. PMID:27304496

  17. Over-expression of NAD kinase in Corynebacterium crenatum and ...

    African Journals Online (AJOL)

    Purpose: To improve the biosynthesis of L-arginine by overexpressing homologous NAD kinase (ppnk) in Corynebacterium crenatum SYPA5-5 and to study its impact in presence of high (HOS) and low oxygen supply (LOS). Methods: A recombinant plasmid (pJC1-tac-ppnK) harboring homologous NAD kinase (ppnk) was ...

  18. Flight controller design of unmanned airplane for radiation monitoring system via structured robust controller design using multiple model approach. Radiation monitoring flight in Namie-machi in Fukushima prefecture

    International Nuclear Information System (INIS)

    Sato, Masayuki; Muraoka, Koji; Hozumi, Koki; Sanada, Yukihisa; Yamada, Tsutomu; Torii, Tatsuo

    2015-01-01

    Due to the tragic accident of radioactive contaminant spread from Fukushima Dai-ichi nuclear power plant, the necessity of unmanned systems for radiation monitoring has been increasing. This paper concerns the flight controller design of an unmanned airplane which has been developed for radiation monitoring around the power plant. The flight controller consists of conventional control elements, i.e. Stability/Control Augmentation System (S/CAS) with PI controllers and guidance loops with PID controllers. The gains in these controllers are designed by minimizing appropriately defined cost functions for several possible models and disturbances to produce structured robust flight controllers. (This method is called as 'multiple model approach'.) Control performance of our flight controller was evaluated through flight tests and a primitive flight of radiation monitoring in Namie-machi in Fukushima prefecture was conducted in Jan. 2014. Flight results are included in this paper. (author)

  19. Targeting NAD+ metabolism in the human malaria parasite Plasmodium falciparum.

    Directory of Open Access Journals (Sweden)

    Jessica K O'Hara

    Full Text Available Nicotinamide adenine dinucleotide (NAD+ is an essential metabolite utilized as a redox cofactor and enzyme substrate in numerous cellular processes. Elevated NAD+ levels have been observed in red blood cells infected with the malaria parasite Plasmodium falciparum, but little is known regarding how the parasite generates NAD+. Here, we employed a mass spectrometry-based metabolomic approach to confirm that P. falciparum lacks the ability to synthesize NAD+ de novo and is reliant on the uptake of exogenous niacin. We characterized several enzymes in the NAD+ pathway and demonstrate cytoplasmic localization for all except the parasite nicotinamidase, which concentrates in the nucleus. One of these enzymes, the P. falciparum nicotinate mononucleotide adenylyltransferase (PfNMNAT, is essential for NAD+ metabolism and is highly diverged from the human homolog, but genetically similar to bacterial NMNATs. Our results demonstrate the enzymatic activity of PfNMNAT in vitro and demonstrate its ability to genetically complement the closely related Escherichia coli NMNAT. Due to the similarity of PfNMNAT to the bacterial enzyme, we tested a panel of previously identified bacterial NMNAT inhibitors and synthesized and screened twenty new derivatives, which demonstrate a range of potency against live parasite culture. These results highlight the importance of the parasite NAD+ metabolic pathway and provide both novel therapeutic targets and promising lead antimalarial compounds.

  20. Enzymology of mammalian NAD metabolism in health and disease.

    Science.gov (United States)

    Magni, Giulio; Orsomando, Giuseppe; Raffelli, Nadia; Ruggieri, Silverio

    2008-05-01

    Mounting evidence attests to the paramount importance of the non-redox NAD functions. Indeed, NAD homeostasis is related to the free radicals-mediated production of reactive oxygen species responsible for irreversible cellular damage in infectious disease, diabetes, inflammatory syndromes, neurodegeneration and cancer. Because the cellular redox status depends on both the absolute concentration of pyridine dinucleotides and their respective ratios of oxidized and reduced forms (i.e., NAD/NADH and NADP/NADPH), it is conceivable that an altered regulation of the synthesis and degradation of NAD impairs the cell redox state and likely contributes to the mechanisms underlying the pathogenesis of the above mentioned diseases. Taking into account the recent appearance in the literature of comprehensive reviews covering different aspects of the significance of NAD metabolism, with particular attention to the enzymes involved in NAD cleavage, this monograph includes the most recent results on NAD biosynthesis in mammals and humans. Due to recent findings on nicotinamide riboside as a nutrient, its inclusion under "niacins" is proposed. Here, the enzymes involved in the de novo and reutilization pathways are overviewed.

  1. The Leishmania nicotinamidase is essential for NAD(+) production and parasite proliferation

    OpenAIRE

    Gazanion, Elodie; Garcia, Deborah; Silvestre, R.; Gérard, C.; Guichou, J. F.; Labesse, G.; Seveno, Martial; Cordeiro-Da-Silva, A.; Ouaissi, A.; Sereno, Denis; Vergnes, Baptiste

    2011-01-01

    NAD(+) is a central cofactor that plays important roles in cellular metabolism and energy production in all living cells. Genomics-based reconstruction of NAD(+) metabolism revealed that Leishmania protozoan parasites are NAD(+) auxotrophs. Consequently, these parasites require assimilating NAD(+) precursors (nicotinamide, nicotinic acid, nicotinamide riboside) from their host environment to synthesize NAD(+) by a salvage pathway. Nicotinamidase is a key enzyme of this salvage pathway that ca...

  2. The Role of NAD+ Depletion in the Mechanisms of Sulfur Mustard-Induced Metabolic Injury

    Science.gov (United States)

    2008-01-01

    reprint. 15. SUBJECT TERMS Cell cultures, glycolysis, HD, keratinocytes, NAD+, niacinamide , sulfur mustard, toxicology 16. SECURITY CLASSIFICATION...NAD+ depletion. To define tl,e role of NAD+ in mu.~tUl·d-induced metabolic injury, we examined the effects ofmustard ± niacinamide on energy metabolism...not earlier, and time- and concentration-dependent glycolytic inhibition and NAD+ depletion as early as 4 hours. Niacinamide partially protected NAD

  3. Novel pathway of NAD metabolism in Aspergillus niger

    International Nuclear Information System (INIS)

    Kuwahara, Masaaki

    1977-01-01

    New steps of NAD metabolism were shown in Aspergillus niger. Radioactive nicotinic acid and nicotinamide were incorporated into nicotinamide ribose diphosphate ribose (NAmRDPR), which had been isolated from the culture filtrate. The enzyme preparation of the mold degraded NAmRDPR to form nicotinamide mononucleotide and nicotinic acid under the neutral and alkaline conditions. In the acid extracts of the mycelia grown on the radioactive precursors, high level of radioactivity was detected on NAD. The experimental results showed that the Preiss-Handler pathway and the NAD cycling system function in the NAD biosynthesis in A. niger. A part of the radioactive precursors was also incorporated into nicotinic acid ribonucleoside, which was thought to be formed from nicotinic acid mononucleotide. (auth.)

  4. TEMPOL increases NAD+ and improves redox imbalance in obese mice

    OpenAIRE

    Yamato, Mayumi; Kawano, Kimika; Yamanaka, Yuki; Saiga, Misako; Yamada, Ken-ichi

    2016-01-01

    Continuous energy conversion is controlled by reduction–oxidation (redox) processes. NAD+ and NADH represent an important redox couple in energy metabolism. 4-Hydroxy-2,2,6,6-tetramethylpiperidine-N-oxyl (TEMPOL) is a redox-cycling nitroxide that promotes the scavenging of several reactive oxygen species (ROS) and is reduced to hydroxylamine by NADH. TEMPOL is also involved in NAD+ production in the ascorbic acid–glutathione redox cycle. We utilized the chemical properties of TEMPOL to invest...

  5. Roles of Nicotinamide Adenine Dinucleotide (NAD+ in Biological Systems

    Directory of Open Access Journals (Sweden)

    Palmiro Poltronieri

    2018-01-01

    Full Text Available NAD+ has emerged as a crucial element in both bioenergetic and signaling pathways since it acts as a key regulator of cellular and organism homeostasis. NAD+ is a coenzyme in redox reactions, a donor of adenosine diphosphate-ribose (ADPr moieties in ADP-ribosylation reactions, a substrate for sirtuins, a group of histone deacetylase enzymes that use NAD+ to remove acetyl groups from proteins; NAD+ is also a precursor of cyclic ADP-ribose, a second messenger in Ca++ release and signaling, and of diadenosine tetraphosphate (Ap4A and oligoadenylates (oligo2′-5′A, two immune response activating compounds. In the biological systems considered in this review, NAD+ is mostly consumed in ADP-ribose (ADPr transfer reactions. In this review the roles of these chemical products are discussed in biological systems, such as in animals, plants, fungi and bacteria. In the review, two types of ADP-ribosylating enzymes are introduced as well as the pathways to restore the NAD+ pools in these systems.

  6. Nii nad tapsidki meie Ferdinandi / Hannes Rumm

    Index Scriptorium Estoniae

    Rumm, Hannes, 1968-

    2007-01-01

    Allar Jõks ei jätka õiguskantslerina. Ilmunud ka Virumaa Teataja 20. dets. 2007, lk. 11 ; Järva Teataja 20. dets. 2007, lk. 2 ; Sakala 21. dets. 2007, lk. 2 ; Põhjarannik 20. dets. 2007, lk. 2 ; Vooremaa 20. dets. 2007, lk. 2 ; Lääne Elu 20. dets. 2007, lk. 2 ; Nädaline 20. dets. 2007, lk. 4 ; Hiiu Leht 28. dets. 2007, lk. 2 ; Pärnu Postimees 9. jaan. 2008, lk. 15 ; Nädalaleht 21. dets. 2007, lk. 4

  7. Loss of NAD Homeostasis Leads to Progressive and Reversible Degeneration of Skeletal Muscle.

    Science.gov (United States)

    Frederick, David W; Loro, Emanuele; Liu, Ling; Davila, Antonio; Chellappa, Karthikeyani; Silverman, Ian M; Quinn, William J; Gosai, Sager J; Tichy, Elisia D; Davis, James G; Mourkioti, Foteini; Gregory, Brian D; Dellinger, Ryan W; Redpath, Philip; Migaud, Marie E; Nakamaru-Ogiso, Eiko; Rabinowitz, Joshua D; Khurana, Tejvir S; Baur, Joseph A

    2016-08-09

    NAD is an obligate co-factor for the catabolism of metabolic fuels in all cell types. However, the availability of NAD in several tissues can become limited during genotoxic stress and the course of natural aging. The point at which NAD restriction imposes functional limitations on tissue physiology remains unknown. We examined this question in murine skeletal muscle by specifically depleting Nampt, an essential enzyme in the NAD salvage pathway. Knockout mice exhibited a dramatic 85% decline in intramuscular NAD content, accompanied by fiber degeneration and progressive loss of both muscle strength and treadmill endurance. Administration of the NAD precursor nicotinamide riboside rapidly ameliorated functional deficits and restored muscle mass despite having only a modest effect on the intramuscular NAD pool. Additionally, lifelong overexpression of Nampt preserved muscle NAD levels and exercise capacity in aged mice, supporting a critical role for tissue-autonomous NAD homeostasis in maintaining muscle mass and function. Copyright © 2016 Elsevier Inc. All rights reserved.

  8. KM3NeT

    CERN Multimedia

    KM3NeT is a large scale next-generation neutrino telescope located in the deep waters of the Mediterranean Sea, optimized for the discovery of galactic neutrino sources emitting in the TeV energy region.

  9. NadN and e (P4) Are Essential for Utilization of NAD and Nicotinamide Mononucleotide but Not Nicotinamide Riboside in Haemophilus influenzae

    Science.gov (United States)

    Kemmer, Gabriele; Reilly, Thomas J.; Schmidt-Brauns, Joachim; Zlotnik, Gary W.; Green, Bruce A.; Fiske, Michael J.; Herbert, Mark; Kraiß, Anita; Schlör, Stefan; Smith, Arnold; Reidl, Joachim

    2001-01-01

    Haemophilus influenzae has an absolute requirement for NAD (factor V) because it lacks almost all the biosynthetic enzymes necessary for the de novo synthesis of that cofactor. Factor V can be provided as either nicotinamide adenosine dinucleotide (NAD), nicotinamide mononucleotide (NMN), or nicotinamide riboside (NR) in vitro, but little is known about the source or the mechanism of uptake of these substrates in vivo. As shown by us earlier, at least two gene products are involved in the uptake of NAD, the outer membrane lipoprotein e (P4), which has phosphatase activity and is encoded by hel, and a periplasmic NAD nucleotidase, encoded by nadN. It has also been observed that the latter gene product is essential for H. influenzae growth on media supplemented with NAD. In this report, we describe the functions and substrates of these two proteins as they act together in an NAD utilization pathway. Data are provided which indicate that NadN harbors not only NAD pyrophosphatase but also NMN 5′-nucleotidase activity. The e (P4) protein is also shown to have NMN 5′-nucleotidase activity, recognizing NMN as a substrate and releasing NR as its product. Insertion mutants of nadN or deletion and site-directed mutants of hel had attenuated growth and a reduced uptake phenotype when NMN served as substrate. A hel and nadN double mutant was only able to grow in the presence of NR, whereas no uptake of NMN was observed. PMID:11395461

  10. NadN and e (P4) are essential for utilization of NAD and nicotinamide mononucleotide but not nicotinamide riboside in Haemophilus influenzae.

    Science.gov (United States)

    Kemmer, G; Reilly, T J; Schmidt-Brauns, J; Zlotnik, G W; Green, B A; Fiske, M J; Herbert, M; Kraiss, A; Schlör, S; Smith, A; Reidl, J

    2001-07-01

    Haemophilus influenzae has an absolute requirement for NAD (factor V) because it lacks almost all the biosynthetic enzymes necessary for the de novo synthesis of that cofactor. Factor V can be provided as either nicotinamide adenosine dinucleotide (NAD), nicotinamide mononucleotide (NMN), or nicotinamide riboside (NR) in vitro, but little is known about the source or the mechanism of uptake of these substrates in vivo. As shown by us earlier, at least two gene products are involved in the uptake of NAD, the outer membrane lipoprotein e (P4), which has phosphatase activity and is encoded by hel, and a periplasmic NAD nucleotidase, encoded by nadN. It has also been observed that the latter gene product is essential for H. influenzae growth on media supplemented with NAD. In this report, we describe the functions and substrates of these two proteins as they act together in an NAD utilization pathway. Data are provided which indicate that NadN harbors not only NAD pyrophosphatase but also NMN 5'-nucleotidase activity. The e (P4) protein is also shown to have NMN 5'-nucleotidase activity, recognizing NMN as a substrate and releasing NR as its product. Insertion mutants of nadN or deletion and site-directed mutants of hel had attenuated growth and a reduced uptake phenotype when NMN served as substrate. A hel and nadN double mutant was only able to grow in the presence of NR, whereas no uptake of NMN was observed.

  11. Effect of X-rays and u.v.-light on the levels of NAD(P), NAD(P)H and hydroxyproline in Pinus silvestris pollen

    International Nuclear Information System (INIS)

    Zelles, L.

    1978-01-01

    Pollen grains of Pinus Silvestris were irradiated with stimulating and inhibiting doses of X-rays and u.v.-light and the levels of NAD(P), NAD(P)H and hydroxyproline determined during tube growth. Pollen grains irradiated with stimulating doses of X-rays and u.v.-light developed longer tubes, while grains irradiated with inhibiting doses of X-rays and u.v.-light developed shorter tubes than the unirradiated controls. After 32 hr of incubation, the levels of NAD(P), NAD(P)H and hydroxyproline were at their lowest compared with unirradiated pollen. In samples with stimulating doses of irradiation NAD(P) reached its maximum earlier than in samples with inhibiting irradiation. The ratio between the concentrations of NAD(P) and NAD(P)H in the irradiated samples was higher than in the unirradiated control. The hydroxyproline content was higher in irradiated than in unirradiated pollen. (author)

  12. Enzymatic Properties of Populus α- and β-NAD-ME Recombinant Proteins

    Directory of Open Access Journals (Sweden)

    Nabil I. Elsheery

    2013-06-01

    Full Text Available Plant mitochondrial NAD-malic enzyme (NAD-ME, which is composed of α- and β-subunits in many species, participates in many plant biosynthetic pathways and in plant respiratory metabolism. However, little is known about the properties of woody plant NAD-MEs. In this study, we analyzed four NAD-ME genes (PtNAD-ME1 through PtNAD-ME4 in the genome of Populus trichocarpa. PtNAD-ME1 and -2 encode putative α-subunits, while PtNAD-ME3 and -4 encode putative β-subunits. The Populus NAD-MEs were expressed in Escherichia coli cells as GST-tagged fusion proteins. Each recombinant GST-PtNAD-ME protein was purified to near homogeneity by glutathione-Sepharose 4B affinity chromatography. Milligram quantities of each native protein were obtained from 1 L bacterial cultures after cleavage of the GST tag. Analysis of the enzymatic properties of these proteins in vitro indicated that α-NAD-MEs are more active than β-NAD-MEs and that α- and β-NAD-MEs presented different kinetic properties (Vmax, kcat and kcat/Km. The effect of different amounts of metabolites on the activities of Populus α- and β-NAD-MEs was assessed in vitro. While none of the metabolites evaluated in our assays activated Populus NAD-ME, oxalacetate and citrate inhibited all α- and β-NAD-MEs and glucose-6-P and fructose inhibited only the α-NAD-MEs.

  13. Enhancing NAD+ salvage metabolism is neuroprotective in a PINK1 model of Parkinson's disease

    Directory of Open Access Journals (Sweden)

    Susann Lehmann

    2017-02-01

    Full Text Available Familial forms of Parkinson's disease (PD caused by mutations in PINK1 are linked to mitochondrial impairment. Defective mitochondria are also found in Drosophila models of PD with pink1 mutations. The co-enzyme nicotinamide adenine dinucleotide (NAD+ is essential for both generating energy in mitochondria and nuclear DNA repair through NAD+-consuming poly(ADP-ribose polymerases (PARPs. We found alterations in NAD+ salvage metabolism in Drosophila pink1 mutants and showed that a diet supplemented with the NAD+ precursor nicotinamide rescued mitochondrial defects and protected neurons from degeneration. Additionally, a mutation of Parp improved mitochondrial function and was neuroprotective in the pink1 mutants. We conclude that enhancing the availability of NAD+ by either the use of a diet supplemented with NAD+ precursors or the inhibition of NAD+-dependent enzymes, such as PARPs, which compete with mitochondria for NAD+, is a viable approach to preventing neurotoxicity associated with mitochondrial defects.

  14. NE2561 and NE2611A - are they different?

    International Nuclear Information System (INIS)

    Huntley, R.; Boas, J.; Kotler, L.; Webb, D.; Stucki, G.

    2000-01-01

    Full text: Evidence is mounting that the nominally identical ionization chamber types NE2561 and NE2611A have significantly different energy dependences. This is revealed by comparing the radiation quality correction factors k q . The factor k q is the ratio of the absorbed dose to water calibration factors (for a particular type of ionization chamber) at radiation quality Q to that for 60 Co. k q values for NE2561 and NE2611A chambers have been compared for various kV and MV X-ray beams at several standards laboratories. Measurements at ARPANSA (Australia) on six NE2561 and five NE2611A show a consistent difference in k q of 1-2% for 16 and 19 MV X-rays. Work at OFMET (Switzerland) has shown similar differences at 6 and 18 MV. No such differences are seen at NPL (UK) - this inconsistency is currently ascribed to differences in the radiation beams. Consistent differences of up to 3% between these two chamber types have been observed at both ARPANSA and NRC (Canada) at the BIPM medium energy X-ray intercomparison qualities between 50 kV and 250 kV. We conclude that the two types of chamber should not be regarded as identical. ARPANSA and several other laboratories in Europe and North America will shortly participate in a Euromet project to be coordinated by OFMET, to investigate high energy X-ray beam quality specifiers. This project will provide additional data that may lead to a better understanding of this anomaly. Copyright (2000) Australasian College of Physical Scientists and Engineers in Medicine

  15. NAD+ biosynthesis ameliorates a zebrafish model of muscular dystrophy.

    Directory of Open Access Journals (Sweden)

    Michelle F Goody

    Full Text Available Muscular dystrophies are common, currently incurable diseases. A subset of dystrophies result from genetic disruptions in complexes that attach muscle fibers to their surrounding extracellular matrix microenvironment. Cell-matrix adhesions are exquisite sensors of physiological conditions and mediate responses that allow cells to adapt to changing conditions. Thus, one approach towards finding targets for future therapeutic applications is to identify cell adhesion pathways that mediate these dynamic, adaptive responses in vivo. We find that nicotinamide riboside kinase 2b-mediated NAD+ biosynthesis, which functions as a small molecule agonist of muscle fiber-extracellular matrix adhesion, corrects dystrophic phenotypes in zebrafish lacking either a primary component of the dystrophin-glycoprotein complex or integrin alpha7. Exogenous NAD+ or a vitamin precursor to NAD+ reduces muscle fiber degeneration and results in significantly faster escape responses in dystrophic embryos. Overexpression of paxillin, a cell adhesion protein downstream of NAD+ in this novel cell adhesion pathway, reduces muscle degeneration in zebrafish with intact integrin receptors but does not improve motility. Activation of this pathway significantly increases organization of laminin, a major component of the extracellular matrix basement membrane. Our results indicate that the primary protective effects of NAD+ result from changes to the basement membrane, as a wild-type basement membrane is sufficient to increase resilience of dystrophic muscle fibers to damage. The surprising result that NAD+ supplementation ameliorates dystrophy in dystrophin-glycoprotein complex- or integrin alpha7-deficient zebrafish suggests the existence of an additional laminin receptor complex that anchors muscle fibers to the basement membrane. We find that integrin alpha6 participates in this pathway, but either integrin alpha7 or the dystrophin-glycoprotein complex is required in conjunction

  16. Enzymatic assay for calmodulins based on plant NAD kinase activity

    Energy Technology Data Exchange (ETDEWEB)

    Harmon, A.C.; Jarrett, H.W.; Cormier, M.J.

    1984-01-01

    NAD kinase with increased sensitivity to calmodulin was purified from pea seedlings (Pisum sativum L., Willet Wonder). Assays for calmodulin based on the activities of NAD kinase, bovine brain cyclic nucleotide phosphodiesterase, and human erythrocyte Ca/sup 2 -/-ATPase were compared for their sensitivities to calmodulin and for their abilities to discriminate between calmodulins from different sources. The activities of the three enzymes were determined in the presence of various concentrations of calmodulins from human erythrocyte, bovine brain, sea pansy (Renilla reniformis), mung bean seed (Vigna radiata L. Wilczek), mushroom (Agaricus bisporus), and Tetrahymena pyriformis. The concentrations of calmodulin required for 50% activation of the NAD kinase (K/sub 0.5/) ranged from 0.520 ng/ml for Tetrahymena to 2.20 ng/ml for bovine brain. The A/sub 0.5/ s ranged from 19.6 ng/ml for bovine brain calmodulin to 73.5 ng/ml for mushroom calmodulin for phosphodiesterase activation. The K/sub 0.5/'s for the activation of Ca/sup 2 +/-ATPase ranged from 36.3 ng/mol for erythrocyte calmodulin to 61.7 ng/ml for mushroom calmodulin. NAD kinase was not stimulated by phosphatidylcholine, phosphatidylserine, cardiolipin, or palmitoleic acid in the absence or presence of Ca/sup 2 +/. Palmitic acid had a slightly stimulatory effect in the presence of Ca/sup 2 +/ (10% of maximum), but no effect in the absence of Ca/sup 2 +/. Palmitoleic acid inhibited the calmodulin-stimulated activity by 50%. Both the NAD kinase assay and radioimmunoassay were able to detect calmodulin in extracts containing low concentrations of calmodulin. Estimates of calmodulin contents of crude homogenates determined by the NAD kinase assay were consistent with amounts obtained by various purification procedures. 30 references, 1 figure, 4 tables.

  17. The reported human NADsyn2 is ammonia-dependent NAD synthetase from a pseudomonad.

    Science.gov (United States)

    Bieganowski, Pawel; Brenner, Charles

    2003-08-29

    Nicotinamide-adenine dinucleotide (NAD+) synthetases catalyze the last step in NAD+ metabolism in the de novo, import, and salvage pathways that originate from tryptophan (or aspartic acid), nicotinic acid, and nicotinamide, respectively, and converge on nicotinic acid mononucleotide. NAD+ synthetase converts nicotinic acid adenine dinucleotide to NAD+ via an adenylylated intermediate. All of the known eukaryotic NAD+ synthetases are glutamine-dependent, hydrolyzing glutamine to glutamic acid to provide the attacking ammonia. In the prokaryotic world, some NAD+ synthetases are glutamine-dependent, whereas others can only use ammonia. Earlier, we noted a perfect correlation between presence of a domain related to nitrilase and glutamine dependence and then proved in the accompanying paper (Bieganowski, P., Pace, H. C., and Brenner, C. (2003) J. Biol. Chem. 278, 33049-33055) that the nitrilase-related domain is an essential, obligate intramolecular, thiol-dependent glutamine amidotransferase in the yeast NAD+ synthetase, Qns1. Independently, human NAD+ synthetase was cloned and shown to depend on Cys-175 for glutamine-dependent but not ammonia-dependent NAD+ synthetase activity. Additionally, it was claimed that a 275 amino acid open reading frame putatively amplified from human glioma cell line LN229 encodes a human ammonia-dependent NAD+ synthetase and this was speculated largely to mediate NAD+ synthesis in human muscle tissues. Here we establish that the so-called NADsyn2 is simply ammonia-dependent NAD+ synthetase from Pseudomonas, which is encoded on an operon with nicotinic acid phosphoribosyltransferase and, in some Pseudomonads, with nicotinamidase.

  18. The NAD/NARB System: Advertising Self-Regulation at Work.

    Science.gov (United States)

    Hays, Robert

    Self-regulation, as defined by the National Advertising Division/National Advertising Review Board (NAD/NARB), is a process whereby the advertising industry regulates itself and turns to the federal government only if the system fails. The NAD/NARB system involves a two-step process: complaints are initially handled by the NAD and then are either…

  19. The NAD(+) precursor nicotinamide riboside decreases exercise performance in rats.

    Science.gov (United States)

    Kourtzidis, Ioannis A; Stoupas, Andreas T; Gioris, Ioannis S; Veskoukis, Aristidis S; Margaritelis, Nikos V; Tsantarliotou, Maria; Taitzoglou, Ioannis; Vrabas, Ioannis S; Paschalis, Vassilis; Kyparos, Antonios; Nikolaidis, Michalis G

    2016-01-01

    Nicotinamide adenine dinucleotide (NAD(+)) and its phosphorylated form (NADP(+)) are key molecules in ubiquitous bioenergetic and cellular signaling pathways, regulating cellular metabolism and homeostasis. Thus, supplementation with NAD(+) and NADP(+) precursors emerged as a promising strategy to gain many and multifaceted health benefits. In this proof-of-concept study, we sought to investigate whether chronic nicotinamide riboside administration (an NAD(+) precursor) affects exercise performance. Eighteen Wistar rats were equally divided in two groups that received either saline vehicle or nicotinamide riboside at a dose of 300 mg/kg body weight/day for 21 days via gavage. At the end of the 21-day administration protocol, both groups performed an incremental swimming performance test. The nicotinamide riboside group showed a tendency towards worse physical performance by 35 % compared to the control group at the final 10 % load (94 ± 53 s for the nicotinamide riboside group and 145 ± 59 s for the control group; P = 0.071). Our results do not confirm the previously reported ergogenic effect of nicotinamide riboside. The potentially negative effect of nicotinamide riboside administration on physical performance may be attributed to the pleiotropic metabolic and redox properties of NAD(+) and NADP(+).

  20. Expression, purification and characterization of Oryza sativa L. NAD ...

    African Journals Online (AJOL)

    Jane

    2011-10-17

    Oct 17, 2011 ... Triton X-100 and 1 mmol/L PMSF). And lysozyme (a final concentration of ... polyacrylamide gel electrophoresis (SDS-PAGE) and proteins were visualized with Coomassie Brilliant ... OsNAD-ME1 that were induced with 1 mmol/L IPTG for different times, and were analyzed on SDS-PAGE. Lane 1, molecular.

  1. Anticancer agent CHS-828 inhibits cellular synthesis of NAD

    DEFF Research Database (Denmark)

    Olesen, U.H.; Christensen, M.K.; Bjorkling, F.

    2008-01-01

    different class. We then showed that nicotinamide protects against CHS-828-mediated cytotoxicity. Finally, we observed that treatment with CHS-828 depletes cellular NAD levels in sensitive cancer cells. In conclusion, these results strongly suggest that, like FK866, CHS-828 kills cancer cells by depleting...

  2. TEMPOL increases NAD+ and improves redox imbalance in obese mice

    Science.gov (United States)

    Yamato, Mayumi; Kawano, Kimika; Yamanaka, Yuki; Saiga, Misako; Yamada, Ken-ichi

    2016-01-01

    Continuous energy conversion is controlled by reduction–oxidation (redox) processes. NAD+ and NADH represent an important redox couple in energy metabolism. 4-Hydroxy-2,2,6,6-tetramethylpiperidine-N-oxyl (TEMPOL) is a redox-cycling nitroxide that promotes the scavenging of several reactive oxygen species (ROS) and is reduced to hydroxylamine by NADH. TEMPOL is also involved in NAD+ production in the ascorbic acid–glutathione redox cycle. We utilized the chemical properties of TEMPOL to investigate the effects of antioxidants and NAD+/NADH modulators on the metabolic imbalance in obese mice. Increases in the NAD+/NADH ratio by TEMPOL ameliorated the metabolic imbalance when combined with a dietary intervention, changing from a high-fat diet to a normal diet. Plasma levels of the superoxide marker dihydroethidium were higher in mice receiving the dietary intervention compared with a control diet, but were normalized with TEMPOL consumption. These findings provide novel insights into redox regulation in obesity. PMID:26942863

  3. TEMPOL increases NAD+ and improves redox imbalance in obese mice

    Directory of Open Access Journals (Sweden)

    Mayumi Yamato

    2016-08-01

    Full Text Available Continuous energy conversion is controlled by reduction–oxidation (redox processes. NAD+ and NADH represent an important redox couple in energy metabolism. 4-Hydroxy-2,2,6,6-tetramethylpiperidine-N-oxyl (TEMPOL is a redox-cycling nitroxide that promotes the scavenging of several reactive oxygen species (ROS and is reduced to hydroxylamine by NADH. TEMPOL is also involved in NAD+ production in the ascorbic acid–glutathione redox cycle. We utilized the chemical properties of TEMPOL to investigate the effects of antioxidants and NAD+/NADH modulators on the metabolic imbalance in obese mice. Increases in the NAD+/NADH ratio by TEMPOL ameliorated the metabolic imbalance when combined with a dietary intervention, changing from a high-fat diet to a normal diet. Plasma levels of the superoxide marker dihydroethidium were higher in mice receiving the dietary intervention compared with a control diet, but were normalized with TEMPOL consumption. These findings provide novel insights into redox regulation in obesity.

  4. TEMPOL increases NAD(+) and improves redox imbalance in obese mice.

    Science.gov (United States)

    Yamato, Mayumi; Kawano, Kimika; Yamanaka, Yuki; Saiga, Misako; Yamada, Ken-Ichi

    2016-08-01

    Continuous energy conversion is controlled by reduction-oxidation (redox) processes. NAD(+) and NADH represent an important redox couple in energy metabolism. 4-Hydroxy-2,2,6,6-tetramethylpiperidine-N-oxyl (TEMPOL) is a redox-cycling nitroxide that promotes the scavenging of several reactive oxygen species (ROS) and is reduced to hydroxylamine by NADH. TEMPOL is also involved in NAD(+) production in the ascorbic acid-glutathione redox cycle. We utilized the chemical properties of TEMPOL to investigate the effects of antioxidants and NAD(+)/NADH modulators on the metabolic imbalance in obese mice. Increases in the NAD(+)/NADH ratio by TEMPOL ameliorated the metabolic imbalance when combined with a dietary intervention, changing from a high-fat diet to a normal diet. Plasma levels of the superoxide marker dihydroethidium were higher in mice receiving the dietary intervention compared with a control diet, but were normalized with TEMPOL consumption. These findings provide novel insights into redox regulation in obesity. Copyright © 2016. Published by Elsevier B.V.

  5. NAD-malic enzymes of Arabidopsis thaliana display distinct kinetic mechanisms that support differences in physiological control.

    Science.gov (United States)

    Tronconi, Marcos A; Gerrard Wheeler, Mariel C; Maurino, Verónica G; Drincovich, María F; Andreo, Carlos S

    2010-09-01

    The Arabidopsis thaliana genome contains two genes encoding NAD-MEs [NAD-dependent malic enzymes; NAD-ME1 (TAIR accession number At4G13560) and NAD-ME2 (TAIR accession number At4G00570)]. The encoded proteins are localized to mitochondria and assemble as homo- and hetero- dimers in vitro and in vivo. In the present work, the kinetic mechanisms of NAD-ME1 and -ME2 homodimers and NAD-MEH (NAD-ME heterodimer) were studied as an approach to understand the contribution of these enzymes to plant physiology. Product-inhibition and substrate-analogue analyses indicated that NAD-ME2 follows a sequential ordered Bi-Ter mechanism, NAD being the leading substrate followed by L-malate. On the other hand, NAD-ME1 and NAD-MEH can bind both substrates randomly. However, NAD-ME1 shows a preferred route that involves the addition of NAD first. As a consequence of the kinetic mechanism, NAD-ME1 showed a partial inhibition by L-malate at low NAD concentrations. The analysis of a protein chimaeric for NAD-ME1 and -ME2 indicated that the first 176 amino acids are associated with the differences observed in the kinetic mechanisms of the enzymes. Furthermore, NAD-ME1, -ME2 and -MEH catalyse the reverse reaction (pyruvate reductive carboxylation) with very low catalytic activity, supporting the notion that these isoforms act only in L-malate oxidation in plant mitochondria. The different kinetic mechanism of each NAD-ME entity suggests that, for a metabolic condition in which the mitochondrial NAD level is low and the L-malate level is high, the activity of NAD-ME2 and/or -MEH would be preferred over that of NAD-ME1.

  6. Three different and tissue-specific NAD-malic enzymes generated by alternative subunit association in Arabidopsis thaliana.

    Science.gov (United States)

    Tronconi, Marcos A; Maurino, Verónica G; Andreo, Carlos S; Drincovich, María F

    2010-04-16

    The Arabidopsis thaliana genome contains two genes encoding the mitochondrial NAD-malic enzyme (NAD-ME), NAD-ME1 (At2g13560) and NAD-ME2 (At4g00570). The characterization of recombinant NAD-ME1 and -2 indicated that both enzymes assemble as active homodimers; however, a heterodimeric enzyme (NAD-MEH) can also be detected by electrophoretic studies. To analyze the metabolic contribution of each enzymatic entity, NAD-MEH was obtained by a co-expression-based recombinant approach, and its kinetic and regulatory properties were analyzed. The three NAD-MEs show similar kinetic properties, although they differ in the regulation by several metabolic effectors. In this regard, whereas fumarate activates NAD-ME1 and CoA activates NAD-ME2, both compounds act synergistically on NAD-MEH activity. The characterization of two chimeric enzymes between NAD-ME1 and -2 allowed specific domains of the primary structure, which are involved in the differential allosteric regulation, to be identified. NAD-ME1 and -2 subunits showed a distinct pattern of accumulation in the separate components of the floral organ. In sepals, the NAD-ME1 subunit is present at a slightly higher proportion than the NAD-ME2 subunit, and thus, NAD-MEH and NAD-ME1 act in concert in this tissue. On the other hand, NAD-ME2 is the only isoform present in anthers. In view of the different properties of NAD-ME1, -2, and -H, we suggest that mitochondrial NAD-ME activity may be regulated by varying native association in vivo, rendering enzymatic entities with distinct allosteric regulation to fulfill specific roles. The presence of three different NAD-ME entities, which originate by alternative associations of two subunits, is suggested to be a novel phenomenon unique to plant mitochondria.

  7. Increasing NAD synthesis in muscle via nicotinamide phosphoribosyltransferase is not sufficient to promote oxidative metabolism.

    Science.gov (United States)

    Frederick, David W; Davis, James G; Dávila, Antonio; Agarwal, Beamon; Michan, Shaday; Puchowicz, Michelle A; Nakamaru-Ogiso, Eiko; Baur, Joseph A

    2015-01-16

    The NAD biosynthetic precursors nicotinamide mononucleotide and nicotinamide riboside are reported to confer resistance to metabolic defects induced by high fat feeding in part by promoting oxidative metabolism in skeletal muscle. Similar effects are obtained by germ line deletion of major NAD-consuming enzymes, suggesting that the bioavailability of NAD is limiting for maximal oxidative capacity. However, because of their systemic nature, the degree to which these interventions exert cell- or tissue-autonomous effects is unclear. Here, we report a tissue-specific approach to increase NAD biosynthesis only in muscle by overexpressing nicotinamide phosphoribosyltransferase, the rate-limiting enzyme in the salvage pathway that converts nicotinamide to NAD (mNAMPT mice). These mice display a ∼50% increase in skeletal muscle NAD levels, comparable with the effects of dietary NAD precursors, exercise regimens, or loss of poly(ADP-ribose) polymerases yet surprisingly do not exhibit changes in muscle mitochondrial biogenesis or mitochondrial function and are equally susceptible to the metabolic consequences of high fat feeding. We further report that chronic elevation of muscle NAD in vivo does not perturb the NAD/NADH redox ratio. These studies reveal for the first time the metabolic effects of tissue-specific increases in NAD synthesis and suggest that critical sites of action for supplemental NAD precursors reside outside of the heart and skeletal muscle. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

  8. Nível de satisfação dos idosos usuários de próteses auditivas doadas pela APAC-NAMI UNIFOR - doi:10.5020/18061230.2005.p7

    Directory of Open Access Journals (Sweden)

    Ana Cristina Martins Batista

    2012-01-01

    Full Text Available A pesquisa objetivou conhecer o nível de satisfação dos pacientes idosos, usuários de próteses auditivas, doadas pela APAC (autorização de atendimento de alta complexidade conveniada com o NAMI – UNIFOR (Universidade Fortaleza. Foram entrevistados, através de aplicação de questionário, 20 pacientes, a partir de 60 anos de idade, de ambos os sexos, beneficiados por doação de AASI (Aparelho de Amplificação Sonora Individual há pelo menos 4 meses. Verificou-se o predomínio do gênero masculino à solicitação de próteses auditivas totalizando 12 pacientes (60%, a presbiacusia revelou-se como causa predominante em 12 pacientes (60%; o tipo de prótese auditiva mais indicada foi a retroauricular em 14 pacientes (70%; observou-se maior rendimento da prótese nos itens relacionados ao uso do mesmo na televisão (60%, conversa (60%, música (60%, conversa na rua (40% e igreja (45%; houve menor satisfação do uso da prótese no shopping center (20% e banco (20%. Quanto às queixas dos usuários, foi referido o uso da prótese em ambientes com sons intensos (25%, seguido do item relacionado ao manuseio dos controles (20%. A aplicação do questionário demonstrou bom nível de satisfação dos pacientes, porém verificou-se que, em determinados ambientes, houve reduzida experienciação por parte dos mesmos, revelando, provavelmente, um estilo de vida mais doméstico.

  9. Rozvoj cyklotrasy Nežárka

    OpenAIRE

    Vondráčková, Jitka

    2010-01-01

    The aim of thesis entitled " The Development of Nežárka Cyclotourism" is using auditing methods to map current state of cycle paths and cycle routes around the river Nežárka,to identify the most appropriate line of the cycle route "Nežárka" and to propose The Cycle route "Nežárka" Master Plan. The theoretical part is focused on bicycle traffic and cycle tourism, particularly cycling infrastructure. The practical part of the thesis includes the line proposal of cycle route "Nežárka", the asses...

  10. Neuronal death induced by misfolded prion protein is due to NAD+ depletion and can be relieved in vitro and in vivo by NAD+ replenishment

    OpenAIRE

    Zhou, Minghai; Ottenberg, Gregory; Sferrazza, Gian Franco; Hubbs, Christopher; Fallahi, Mohammad; Rumbaugh, Gavin; Brantley, Alicia F.; Lasmézas, Corinne I.

    2015-01-01

    The mechanisms by which misfolded proteins trigger neurodegeneration remain unclear. Zhou et al. show that the misfolded prion protein TPrP triggers abnormal autophagy activation and neuronal death via NAD + depletion resulting from excessive PARP1-independent ADP-ribosylation. NAD + replenishment rescues prion protein-damaged neurons, suggesting neuroprotective potential in prion-mediated neurodegenerative diseases.

  11. The effector AvrRxo1 phosphorylates NAD in planta.

    Directory of Open Access Journals (Sweden)

    Teja Shidore

    2017-06-01

    Full Text Available Gram-negative bacterial pathogens of plants and animals employ type III secreted effectors to suppress innate immunity. Most characterized effectors work through modification of host proteins or transcriptional regulators, although a few are known to modify small molecule targets. The Xanthomonas type III secreted avirulence factor AvrRxo1 is a structural homolog of the zeta toxin family of sugar-nucleotide kinases that suppresses bacterial growth. AvrRxo1 was recently reported to phosphorylate the central metabolite and signaling molecule NAD in vitro, suggesting that the effector might enhance bacterial virulence on plants through manipulation of primary metabolic pathways. In this study, we determine that AvrRxo1 phosphorylates NAD in planta, and that its kinase catalytic sites are necessary for its toxic and resistance-triggering phenotypes. A global metabolomics approach was used to independently identify 3'-NADP as the sole detectable product of AvrRxo1 expression in yeast and bacteria, and NAD kinase activity was confirmed in vitro. 3'-NADP accumulated upon transient expression of AvrRxo1 in Nicotiana benthamiana and in rice leaves infected with avrRxo1-expressing strains of X. oryzae. Mutation of the catalytic aspartic acid residue D193 abolished AvrRxo1 kinase activity and several phenotypes of AvrRxo1, including toxicity in yeast, bacteria, and plants, suppression of the flg22-triggered ROS burst, and ability to trigger an R gene-mediated hypersensitive response. A mutation in the Walker A ATP-binding motif abolished the toxicity of AvrRxo1, but did not abolish the 3'-NADP production, virulence enhancement, ROS suppression, or HR-triggering phenotypes of AvrRxo1. These results demonstrate that a type III effector targets the central metabolite and redox carrier NAD in planta, and that this catalytic activity is required for toxicity and suppression of the ROS burst.

  12. Badania nad komunikacją międzykulturową

    DEFF Research Database (Denmark)

    Wilczewski, Michał; Søderberg, Anne-Marie

    2017-01-01

    jako badanie narracyjne. Proponujemy, by podejście narracyjne zostało wykorzystane w badaniach nad komunikacją międzykulturową, gdyż oferuje narzędzia dające dostęp do sposobów, w jakie uczestnicy komunikacji opowiadają o swoich międzykulturowych doświadczeniach, do refleksji na ich temat, więc metoda...

  13. NAD+ Modulates p53 DNA Binding Specificity and Function

    Science.gov (United States)

    McLure, Kevin G.; Takagi, Masatoshi; Kastan, Michael B.

    2004-01-01

    DNA damage induces p53 DNA binding activity, which affects tumorigenesis, tumor responses to therapies, and the toxicities of cancer therapies (B. Vogelstein, D. Lane, and A. J. Levine, Nature 408:307-310, 2000; K. H. Vousden and X. Lu, Nat. Rev. Cancer 2:594-604, 2002). Both transcriptional and transcription-independent activities of p53 contribute to DNA damage-induced cell cycle arrest, apoptosis, and aneuploidy prevention (M. B. Kastan et al., Cell 71:587-597, 1992; K. H. Vousden and X. Lu, Nat. Rev. Cancer 2:594-604, 2002). Small-molecule manipulation of p53 DNA binding activity has been an elusive goal, but here we show that NAD+ binds to p53 tetramers, induces a conformational change, and modulates p53 DNA binding specificity in vitro. Niacinamide (vitamin B3) increases the rate of intracellular NAD+ synthesis, alters radiation-induced p53 DNA binding specificity, and modulates activation of a subset of p53 transcriptional targets. These effects are likely due to a direct effect of NAD+ on p53, as a molecule structurally related to part of NAD+, TDP, also inhibits p53 DNA binding, and the TDP precursor, thiamine (vitamin B1), inhibits intracellular p53 activity. Niacinamide and thiamine affect two p53-regulated cellular responses to ionizing radiation: rereplication and apoptosis. Thus, niacinamide and thiamine form a novel basis for the development of small molecules that affect p53 function in vivo, and these results suggest that changes in cellular energy metabolism may regulate p53. PMID:15509798

  14. Nicotinamide riboside kinase structures reveal new pathways to NAD+.

    Directory of Open Access Journals (Sweden)

    Wolfram Tempel

    2007-10-01

    Full Text Available The eukaryotic nicotinamide riboside kinase (Nrk pathway, which is induced in response to nerve damage and promotes replicative life span in yeast, converts nicotinamide riboside to nicotinamide adenine dinucleotide (NAD+ by phosphorylation and adenylylation. Crystal structures of human Nrk1 bound to nucleoside and nucleotide substrates and products revealed an enzyme structurally similar to Rossmann fold metabolite kinases and allowed the identification of active site residues, which were shown to be essential for human Nrk1 and Nrk2 activity in vivo. Although the structures account for the 500-fold discrimination between nicotinamide riboside and pyrimidine nucleosides, no enzyme feature was identified to recognize the distinctive carboxamide group of nicotinamide riboside. Indeed, nicotinic acid riboside is a specific substrate of human Nrk enzymes and is utilized in yeast in a novel biosynthetic pathway that depends on Nrk and NAD+ synthetase. Additionally, nicotinic acid riboside is utilized in vivo by Urh1, Pnp1, and Preiss-Handler salvage. Thus, crystal structures of Nrk1 led to the identification of new pathways to NAD+.

  15. Nicotinamide riboside kinase structures reveal new pathways to NAD+.

    Science.gov (United States)

    Tempel, Wolfram; Rabeh, Wael M; Bogan, Katrina L; Belenky, Peter; Wojcik, Marzena; Seidle, Heather F; Nedyalkova, Lyudmila; Yang, Tianle; Sauve, Anthony A; Park, Hee-Won; Brenner, Charles

    2007-10-02

    The eukaryotic nicotinamide riboside kinase (Nrk) pathway, which is induced in response to nerve damage and promotes replicative life span in yeast, converts nicotinamide riboside to nicotinamide adenine dinucleotide (NAD+) by phosphorylation and adenylylation. Crystal structures of human Nrk1 bound to nucleoside and nucleotide substrates and products revealed an enzyme structurally similar to Rossmann fold metabolite kinases and allowed the identification of active site residues, which were shown to be essential for human Nrk1 and Nrk2 activity in vivo. Although the structures account for the 500-fold discrimination between nicotinamide riboside and pyrimidine nucleosides, no enzyme feature was identified to recognize the distinctive carboxamide group of nicotinamide riboside. Indeed, nicotinic acid riboside is a specific substrate of human Nrk enzymes and is utilized in yeast in a novel biosynthetic pathway that depends on Nrk and NAD+ synthetase. Additionally, nicotinic acid riboside is utilized in vivo by Urh1, Pnp1, and Preiss-Handler salvage. Thus, crystal structures of Nrk1 led to the identification of new pathways to NAD+.

  16. External NAD(P)H dehydrogenases in Acanthamoeba castellanii mitochondria.

    Science.gov (United States)

    Antos-Krzeminska, Nina; Jarmuszkiewicz, Wieslawa

    2014-09-01

    The mitochondrial respiratory chain of plants and some fungi contains multiple rotenone-insensitive NAD(P)H dehydrogenases, of which at least two are located on the outer surface of the inner membrane (i.e., external NADH and external NADPH dehydrogenases). Annotated sequences of the putative alternative NAD(P)H dehydrogenases of the protozoan Acanthamoeba castellanii demonstrated similarity to plant and fungal sequences. We also studied activity of these dehydrogenases in isolated A. castellanii mitochondria. External NADPH oxidation was observed for the first time in protist mitochondria. The coupling parameters were similar for external NADH oxidation and external NADPH oxidation, indicating similar efficiencies of ATP synthesis. Both external NADH oxidation and external NADPH oxidation had an optimal pH of 6.8 independent of relevant ubiquinol-oxidizing pathways, the cytochrome pathway or a GMP-stimulated alternative oxidase. The maximal oxidizing activity with external NADH was almost double that with external NADPH. However, a lower Michaelis constant (K(M)) value for external NADPH oxidation was observed compared to that for external NADH oxidation. Stimulation by Ca(2+) was approximately 10 times higher for external NADPH oxidation, while NADH dehydrogenase(s) appeared to be slightly dependent on Ca(2+). Our results indicate that external NAD(P)H dehydrogenases similar to those in plant and fungal mitochondria function in mitochondria of A. castellanii. Copyright © 2014 Elsevier GmbH. All rights reserved.

  17. Biosynthesis of NAD from nicotinic acid and nicotinamide by resting cells of Arthrobacter globiformis

    International Nuclear Information System (INIS)

    Kuwahara, Masaaki

    1978-01-01

    Isotopically labeled nicotinic acid and nicotinamide were incorporated into the metabolites of nicotinic acid-dependent pathway (Preiss-Handler pathway) of the NAD biosynthesis by resting cells of Arthrobacter globiformis. Azaserine and adenosine markedly stimulated the accumulation of NAD in the cells. Radioactive nicotinic acid and nicotinamide were also incorporated into an unknown compound when the cells were incubated in the presence of azaserine. Cell-free extract of the organism showed the NAD synthetase activity, which required ammonium ion and ATP for the amidation of deamido-NAD. Adenosine inhibited the enzyme activity. The organism possessed nicotinamidase, suggesting deamidation is the first step in the biosynthesis of NAD from nicotinamide. The activity was inhibited by NAD, NADP and NMN. (auth.)

  18. NAD+Intermediates: The Biology and Therapeutic Potential of NMN and NR.

    Science.gov (United States)

    Yoshino, Jun; Baur, Joseph A; Imai, Shin-Ichiro

    2017-12-14

    Research on the biology of NAD + has been gaining momentum, providing many critical insights into the pathogenesis of age-associated functional decline and diseases. In particular, two key NAD + intermediates, nicotinamide riboside (NR) and nicotinamide mononucleotide (NMN), have been extensively studied over the past several years. Supplementing these NAD + intermediates has shown preventive and therapeutic effects, ameliorating age-associated pathophysiologies and disease conditions. Although the pharmacokinetics and metabolic fates of NMN and NR are still under intensive investigation, these NAD + intermediates can exhibit distinct behavior, and their fates appear to depend on the tissue distribution and expression levels of NAD + biosynthetic enzymes, nucleotidases, and presumptive transporters for each. A comprehensive concept that connects NAD + metabolism to the control of aging and longevity in mammals has been proposed, and the stage is now set to test whether these exciting preclinical results can be translated to improve human health. Copyright © 2017 Elsevier Inc. All rights reserved.

  19. Therapeutic Potential of NAD-Boosting Molecules: The In Vivo Evidence.

    Science.gov (United States)

    Rajman, Luis; Chwalek, Karolina; Sinclair, David A

    2018-03-06

    Nicotinamide adenine dinucleotide (NAD), the cell's hydrogen carrier for redox enzymes, is well known for its role in redox reactions. More recently, it has emerged as a signaling molecule. By modulating NAD + -sensing enzymes, NAD + controls hundreds of key processes from energy metabolism to cell survival, rising and falling depending on food intake, exercise, and the time of day. NAD + levels steadily decline with age, resulting in altered metabolism and increased disease susceptibility. Restoration of NAD + levels in old or diseased animals can promote health and extend lifespan, prompting a search for safe and efficacious NAD-boosting molecules that hold the promise of increasing the body's resilience, not just to one disease, but to many, thereby extending healthy human lifespan. Copyright © 2018 Elsevier Inc. All rights reserved.

  20. Upregulation of mitochondrial NAD+ levels impairs the clonogenicity of SSEA1+ glioblastoma tumor-initiating cells

    OpenAIRE

    Son, Myung Jin; Ryu, Jae-Sung; Kim, Jae Yun; Kwon, Youjeong; Chung, Kyung-Sook; Mun, Seon Ju; Cho, Yee Sook

    2017-01-01

    Emerging evidence has emphasized the importance of cancer therapies targeting an abnormal metabolic state of tumor-initiating cells (TICs) in which they retain stem cell-like phenotypes and nicotinamide adenine dinucleotide (NAD+) metabolism. However, the functional role of NAD+ metabolism in regulating the characteristics of TICs is not known. In this study, we provide evidence that the mitochondrial NAD+ levels affect the characteristics of glioma-driven SSEA1+ TICs, including clonogenic gr...

  1. Yeast Life-Span Extension by Calorie Restriction Is Independent of NAD Fluctuation

    Science.gov (United States)

    Neves, Ana Rute; Lavu, Siva; Medvedik, Oliver; Taylor, Christopher; Howitz, Konrad T.; Santos, Helena; Sinclair, David A.

    2016-01-01

    Calorie restriction (CR) slows aging in numerous species. In the yeast Saccharomyces cerevisiae, this effect requires Sir2, a conserved NAD+-dependent deacetylase. We report that CR reduces nuclear NAD+ levels in vivo. Moreover, the activity of Sir2 and its human homologue SIRT1 are not affected by physiological alterations in the NAD+:NADH ratio. These data implicate alternate mechanisms of Sir2 regulation by CR. PMID:14605207

  2. Novel Type II and Monomeric NAD+ Specific Isocitrate Dehydrogenases: Phylogenetic Affinity, Enzymatic Characterization, and Evolutionary Implication

    OpenAIRE

    Wang, Peng; Lv, Changqi; Zhu, Guoping

    2015-01-01

    NAD+ use is an ancestral trait of isocitrate dehydrogenase (IDH), and the NADP+ phenotype arose through evolution as an ancient adaptation event. However, no NAD+-specific IDHs have been found among type II IDHs and monomeric IDHs. In this study, novel type II homodimeric NAD-IDHs from Ostreococcus lucimarinus CCE9901 IDH (OlIDH) and Micromonas sp. RCC299 (MiIDH), and novel monomeric NAD-IDHs from Campylobacter sp. FOBRC14 IDH (CaIDH) and Campylobacter curvus (CcIDH) were reported for the fir...

  3. The effect of gamma radiation and neocarzinostatin on NAD and ATP levels in mouse leukaemia cells

    International Nuclear Information System (INIS)

    Goodwin, P.M.; Lewis, P.J.; Davies, M.I.; Skidmore, C.J.; Shall, S.

    1978-01-01

    When mouse leukaemia cells are treated with γ-radiation or neocarzinostatin the intracelluiar NAD and ATP levels fall rapidly. It is shown that the ATP response is a consequence of the decreased NAD level. It is suggested that this low NAD level results in decreased glycolytic activity and that there is a subsequent accumulation of phosphorylated sugars associated with the fall in ATP. Under these extreme conditions, therefore, the NAD level probably regulates the rate of glycolysis in cells which are utilising a rapidly metabolisable sugar as their energy source. (Auth.)

  4. Engineering NAD+ availability for Escherichia coli whole-cell biocatalysis: a case study for dihydroxyacetone production.

    Science.gov (United States)

    Zhou, Yongjin J; Yang, Wei; Wang, Lei; Zhu, Zhiwei; Zhang, Sufang; Zhao, Zongbao K

    2013-11-09

    Whole-cell redox biocatalysis has been intensively explored for the production of valuable compounds because excellent selectivity is routinely achieved. Although the cellular cofactor level, redox state and the corresponding enzymatic activity are expected to have major effects on the performance of the biocatalysts, our ability remains limited to predict the outcome upon variation of those factors as well as the relationship among them. In order to investigate the effects of cofactor availability on whole-cell redox biocatalysis, we devised recombinant Escherichia coli strains for the production of dihydroxyacetone (DHA) catalyzed by the NAD+-dependent glycerol dehydrogenase (GldA). In this model system, a water-forming NAD+ oxidase (NOX) and a NAD+ transporter (NTT4) were also co-expressed for cofactor regeneration and extracellular NAD+ uptake, respectively. We found that cellular cofactor level, NAD+/NADH ratio and NOX activity were not only strain-dependent, but also growth condition-dependent, leading to significant differences in specific DHA titer among different whole-cell biocatalysts. The host E. coli DH5α had the highest DHA specific titer of 0.81 g/gDCW with the highest NAD+/NADH ratio of 6.7 and NOX activity of 3900 U. The biocatalyst had a higher activity when induced with IPTG at 37°C for 8 h compared with those at 30°C for 8 h and 18 h. When cells were transformed with the ntt4 gene, feeding NAD+ during the cell culture stage increased cellular NAD(H) level by 1.44 fold and DHA specific titer by 1.58 fold to 2.13 g/gDCW. Supplementing NAD+ during the biotransformation stage was also beneficial to cellular NAD(H) level and DHA production, and the highest DHA productivity reached 0.76 g/gDCW/h. Cellular NAD(H) level, NAD+/NADH ratio, and NOX and GldA activity dropped over time during the biotransformation process. High NAD+/NADH ratio driving by NOX was very important for DHA production. Once cofactor was efficiently cycled, high cellular

  5. Age related changes in NAD+ metabolism oxidative stress and Sirt1 activity in wistar rats.

    Directory of Open Access Journals (Sweden)

    Nady Braidy

    2011-04-01

    Full Text Available The cofactor nicotinamide adenine dinucleotide (NAD+ has emerged as a key regulator of metabolism, stress resistance and longevity. Apart from its role as an important redox carrier, NAD+ also serves as the sole substrate for NAD-dependent enzymes, including poly(ADP-ribose polymerase (PARP, an important DNA nick sensor, and NAD-dependent histone deacetylases, Sirtuins which play an important role in a wide variety of processes, including senescence, apoptosis, differentiation, and aging. We examined the effect of aging on intracellular NAD+ metabolism in the whole heart, lung, liver and kidney of female wistar rats. Our results are the first to show a significant decline in intracellular NAD+ levels and NAD:NADH ratio in all organs by middle age (i.e.12 months compared to young (i.e. 3 month old rats. These changes in [NAD(H] occurred in parallel with an increase in lipid peroxidation and protein carbonyls (o- and m- tyrosine formation and decline in total antioxidant capacity in these organs. An age dependent increase in DNA damage (phosphorylated H2AX was also observed in these same organs. Decreased Sirt1 activity and increased acetylated p53 were observed in organ tissues in parallel with the drop in NAD+ and moderate over-expression of Sirt1 protein. Reduced mitochondrial activity of complex I-IV was also observed in aging animals, impacting both redox status and ATP production. The strong positive correlation observed between DNA damage associated NAD+ depletion and Sirt1 activity suggests that adequate NAD+ concentrations may be an important longevity assurance factor.

  6. Hepatic NAD(+) deficiency as a therapeutic target for non-alcoholic fatty liver disease in ageing.

    Science.gov (United States)

    Zhou, Can-Can; Yang, Xi; Hua, Xia; Liu, Jian; Fan, Mao-Bing; Li, Guo-Qiang; Song, Jie; Xu, Tian-Ying; Li, Zhi-Yong; Guan, Yun-Feng; Wang, Pei; Miao, Chao-Yu

    2016-08-01

    Ageing is an important risk factor of non-alcoholic fatty liver disease (NAFLD). Here, we investigated whether the deficiency of nicotinamide adenine dinucleotide (NAD(+) ), a ubiquitous coenzyme, links ageing with NAFLD. Hepatic concentrations of NAD(+) , protein levels of nicotinamide phosphoribosyltransferase (NAMPT) and several other critical enzymes regulating NAD(+) biosynthesis, were compared in middle-aged and aged mice or patients. The influences of NAD(+) decline on the steatosis and steatohepatitis were evaluated in wild-type and H247A dominant-negative, enzymically-inactive NAMPT transgenic mice (DN-NAMPT) given normal or high-fat diet (HFD). Hepatic NAD(+) level decreased in aged mice and humans. NAMPT-controlled NAD(+) salvage, but not de novo biosynthesis pathway, was compromised in liver of elderly mice and humans. Given normal chow, middle-age DN-NAMPT mice displayed systemic NAD(+) reduction and had moderate NAFLD phenotypes, including lipid accumulation, enhanced oxidative stress, triggered inflammation and impaired insulin sensitivity in liver. All these NAFLD phenotypes, especially release of pro-inflammatory factors, Kupffer cell accumulation, monocytes infiltration, NLRP3 inflammasome pathway and hepatic fibrosis (Masson's staining and α-SMA staining), deteriorated further under HFD challenge. Oral administration of nicotinamide riboside, a natural NAD(+) precursor, completely corrected these NAFLD phenotypes induced by NAD(+) deficiency alone or HFD, whereas adenovirus-mediated SIRT1 overexpression only partially rescued these phenotypes. These results provide the first evidence that ageing-associated NAD(+) deficiency is a critical risk factor for NAFLD, and suggest that supplementation with NAD(+) substrates may be a promising therapeutic strategy to prevent and treat NAFLD. © 2016 The British Pharmacological Society.

  7. The study of NAD-malic enzyme in Amaranthus cruentus L. under drought.

    Science.gov (United States)

    Babayev, Hasan; Mehvaliyeva, Ulduza; Aliyeva, Minakhanym; Feyziyev, Yashar; Guliyev, Novruz

    2014-08-01

    Decarboxylating NAD-malate dehydrogenase (NAD-malic enzyme, NAD-ME, EC 1.1.1.39) has been investigated under a long-term drought during pre-anthesis, anthesis and seed-formation phases of ontogenesis of a NAD-ME type C4 plant Amaranthus cruentus L. using cytosol, chloroplast and mitochondrial fractions of mesophyll (M) and bundle sheath (BS) cells. We detected several molecular forms of NAD-ME with different subcellular localization patterns in the studied phases of amaranth ontogenesis. However, no enzyme activity was observed experimentally in chloroplasts of M and BS cells. In the pre-anthesis phase NAD-ME isoform with molecular weight of ∼115 kDa was found in cytosol of M and BS cells of control and drought-exposed plants. One of NAD-ME isoforms with molecular weight of 110 kDa was located in mitochondria of BS cells of control and drought-exposed plants, and a new isoform of ∼121 kDa was formed in mitochondria of BS cells under the influence of drought. After resuming watering this isoform (∼121 kDa) disappeared again. Approximately 90.6% and 9.4% of the total NAD-ME activity were localized in mitochondrial stroma and cytosol of BS cells, respectively, while in mesophyll cells 100% activity was found in cytosol fractions. The reaction catalyzed by NAD-ME follows Michaelis-Menten equation. NAD(+), l-malate and Mn(2+) activate this enzyme in mitochondria. Appearance of the ∼121 kDa isoform of NAD-ME in the mitochondrial fraction of BS cells under drought and its disappearance after resuming watering could be attributed to one of the protection functions of plants. Copyright © 2014 Elsevier Masson SAS. All rights reserved.

  8. Monitoring of mercury concentration in atmosphere in Usti nad Labem

    International Nuclear Information System (INIS)

    Synek, V.; Baloch, T.; Otcenasek, J.; Kremlova, S.; Subrt, P.

    2007-01-01

    This study elaborates the observation of mercury pollution of the atmosphere in the city of Usti nad Labem. The biggest source of the polluting mercury in Usti nad Labem is the chlor-alkali production in the factory of Spolchemie Inc. The method of mercury determination applied is based on capturing the mercury contented in a volume of the air on an amalgamator and measuring the mercury by an atomic absorption spectrometer (Perkin -Elmer 4100ZL) equipped with a special adapter after a thermal release of the mercury from the amalgamator. The basic characteristics of this method were evaluated; e.g. the limit of detection and limit of determination are, respectively, 0.43 and 1.4 ng/m 3 , the relative expanded uncertainty is 28 %. The work gives results of long-term (1998-2006) observations in a few localities in Usti nad Labem situated in various distances from the mercury source (e.g. means of 28.6 and 14.1 ng/m3 were obtained, respectively, in places 350 and 700 m far from the electrolysis plant) and also in a different city (Duchcov). The cases with a higher mercury concentration are very frequent so the sets of the obtained results have lognormal distributions. This study statistically compares the total level and variability of the mercury concentrations in the time series. It also investigates their trends, correlations between them and meteorological influences upon the levels of mercury concentration in the air. The effect of the mercury emission from the chlor-alkali plant is dominant. It as the only factor determines when the cases with a high mercury concentration in the atmosphere occur. (author)

  9. Catalases are NAD(PH-dependent tellurite reductases.

    Directory of Open Access Journals (Sweden)

    Iván L Calderón

    2006-12-01

    Full Text Available Reactive oxygen species damage intracellular targets and are implicated in cancer, genetic disease, mutagenesis, and aging. Catalases are among the key enzymatic defenses against one of the most physiologically abundant reactive oxygen species, hydrogen peroxide. The well-studied, heme-dependent catalases accelerate the rate of the dismutation of peroxide to molecular oxygen and water with near kinetic perfection. Many catalases also bind the cofactors NADPH and NADH tenaciously, but, surprisingly, NAD(PH is not required for their dismutase activity. Although NAD(PH protects bovine catalase against oxidative damage by its peroxide substrate, the catalytic role of the nicotinamide cofactor in the function of this enzyme has remained a biochemical mystery to date. Anions formed by heavy metal oxides are among the most highly reactive, natural oxidizing agents. Here, we show that a natural isolate of Staphylococcus epidermidis resistant to tellurite detoxifies this anion thanks to a novel activity of its catalase, and that a subset of both bacterial and mammalian catalases carry out the NAD(PH-dependent reduction of soluble tellurite ion (TeO(3(2- to the less toxic, insoluble metal, tellurium (Te(o, in vitro. An Escherichia coli mutant defective in the KatG catalase/peroxidase is sensitive to tellurite, and expression of the S. epidermidis catalase gene in a heterologous E. coli host confers increased resistance to tellurite as well as to hydrogen peroxide in vivo, arguing that S. epidermidis catalase provides a physiological line of defense against both of these strong oxidizing agents. Kinetic studies reveal that bovine catalase reduces tellurite with a low Michaelis-Menten constant, a result suggesting that tellurite is among the natural substrates of this enzyme. The reduction of tellurite by bovine catalase occurs at the expense of producing the highly reactive superoxide radical.

  10. Lucius Annaeus SENECA: On Providence (Why the Good Suffer Misfortunes Although There Is a Providence (translation

    Directory of Open Access Journals (Sweden)

    Dragica Fabjan

    2004-12-01

    Full Text Available Vprašal si me, Lucilij, kako to, da se dobrim ljudem pripeti toliko hudega, če pa svetu vlada Previdnost. O tem bi bilo primerneje govoriti v posebni razpravi, kjer bi skušal dokazati, da ves svet vodi božja Previdnost in da Bog bdi nad nami. Tebi pa je ljubše, če iz celote iztrgam le delček in ovržem en sam ugovor, ne da bi se dotaknil spornega vprašanja. Zato bom zagovarjal ravnanje bogov, kar ni tako težko.

  11. Marlène Mocquet

    OpenAIRE

    Saudrais, Anthony

    2012-01-01

    Les œuvres (de 2009 à 2011) ici réunies réinterprètent le divorce des idées reçues entre l’art figuratif et l’art abstrait. La retranscription d’une conversation, justement intitulée « Les Gigognes », entre l’artiste et Gaëlle Rageot-Deshayes, aide à cerner les références de Marlène Mocquet, la matérialité de ses œuvres, « le principe de la poupée russe [qui] gouverne [ses] images » (p.12) ou encore le jeu des titres et du récit imaginaire [« La Fraise dans ma bouche » (2009) ; « Lire l’oisea...

  12. Znečištění ovzduší v Praze, Teplicích a v Prachaticích v uplynulých 15 letech. Porovnání dat a úvaha nad změnami a trendy

    Czech Academy of Sciences Publication Activity Database

    Beneš, I.; Skorkovský, J.; Novák, J.; Šrám, Radim

    5-6, - (2010), s. 18-23 ISSN 1211-0337 R&D Projects: GA MŽP(CZ) SP/1B3/8/08; GA MŽP(CZ) SP/1B3/50/07 Institutional research plan: CEZ:AV0Z50390512 Keywords : air pollution * sulfur dioxide * carbon monoxide Subject RIV: DN - Health Impact of the Environment Quality

  13. MiniBooNE Oscillation Results

    International Nuclear Information System (INIS)

    Djurcic, Zelimir

    2009-01-01

    These proceedings summarize the MiniBooNE ν μ → ν e results, describe the first (bar ν) μ → (bar ν) e result, and current analysis effort with the NuMI neutrinos detected in the miniBooNE detector

  14. MiniBooNE overview and status

    Indian Academy of Sciences (India)

    physics pp. 611-614. MiniBooNE overview and status. P KAsPER (for the BooNE Collaboration). Fermilab, P.O. Box 500, Batavia, IL 60510-0500, USA. Abstract. Recent discoveries in the neutrino sector have opened a new frontier in high- energy physics and cosmology. Evidence from neutrino oscillation experiments from.

  15. The Leishmania nicotinamidase is essential for NAD+ production and parasite proliferation.

    Science.gov (United States)

    Gazanion, E; Garcia, D; Silvestre, R; Gérard, C; Guichou, J F; Labesse, G; Seveno, M; Cordeiro-Da-Silva, A; Ouaissi, A; Sereno, D; Vergnes, B

    2011-10-01

    NAD+ is a central cofactor that plays important roles in cellular metabolism and energy production in all living cells. Genomics-based reconstruction of NAD+ metabolism revealed that Leishmania protozoan parasites are NAD+ auxotrophs. Consequently, these parasites require assimilating NAD+ precursors (nicotinamide, nicotinic acid, nicotinamide riboside) from their host environment to synthesize NAD+ by a salvage pathway. Nicotinamidase is a key enzyme of this salvage pathway that catalyses conversion of nicotinamide (NAm) to nicotinic acid (Na), and that is absent in higher eukaryotes. We present here the biochemical and functional characterizations of the Leishmania infantum nicotinamidase (LiPNC1). Generation of Lipnc1 null mutants leads to a decrease in NAD+ content, associated with a metabolic shutdown-like phenotype with an extensive lag phase of growth. Both phenotypes could be rescued by an add-back construct or by addition of exogenous Na. In addition, Lipnc1 null mutants were unable to establish a sustained infection in a murine experimental model. Altogether, these results illustrate that NAD+ homeostasis is a fundamental component of Leishmania biology and virulence, and that NAm constitutes its main NAD+ source in the mammalian host. The crystal structure of LiPNC1 we solved allows now the design of rational inhibitors against this new promising therapeutic target. © 2011 Blackwell Publishing Ltd.

  16. Nicotinamide Riboside Is a Major NAD+ Precursor Vitamin in Cow Milk.

    Science.gov (United States)

    Trammell, Samuel Aj; Yu, Liping; Redpath, Philip; Migaud, Marie E; Brenner, Charles

    2016-05-01

    Nicotinamide riboside (NR) is a recently discovered NAD(+) precursor vitamin with a unique biosynthetic pathway. Although the presence of NR in cow milk has been known for more than a decade, the concentration of NR with respect to the other NAD(+) precursors was unknown. We aimed to determine NAD(+) precursor vitamin concentration in raw samples of milk from individual cows and from commercially available cow milk. LC tandem mass spectrometry and isotope dilution technologies were used to quantify NAD(+) precursor vitamin concentration and to measure NR stability in raw and commercial milk. Nuclear magnetic resonance (NMR) spectroscopy was used to test for NR binding to substances in milk. Cow milk typically contained ∼12 μmol NAD(+) precursor vitamins/L, of which 60% was present as nicotinamide and 40% was present as NR. Nicotinic acid and other NAD(+) metabolites were below the limits of detection. Milk from samples testing positive for Staphylococcus aureus contained lower concentrations of NR (Spearman ρ = -0.58, P = 0.014), and NR was degraded by S. aureus Conventional milk contained more NR than milk sold as organic. Nonetheless, NR was stable in organic milk and exhibited an NMR spectrum consistent with association with a protein fraction in skim milk. NR is a major NAD(+) precursor vitamin in cow milk. Control of S. aureus may be important to preserve the NAD(+) precursor vitamin concentration of milk. © 2016 American Society for Nutrition.

  17. 33 CFR 110.168 - Hampton Roads, Virginia and adjacent waters (Datum: NAD 83).

    Science.gov (United States)

    2010-07-01

    ... 33 Navigation and Navigable Waters 1 2010-07-01 2010-07-01 false Hampton Roads, Virginia and adjacent waters (Datum: NAD 83). 110.168 Section 110.168 Navigation and Navigable Waters COAST GUARD..., Virginia and adjacent waters (Datum: NAD 83). (a) Anchorage Grounds—(1) Anchorage A [Naval Anchorage]. The...

  18. Deficiency of the Mitochondrial NAD Kinase Causes Stress-Induced Hepatic Steatosis in Mice

    NARCIS (Netherlands)

    Zhang, Kezhong; Kim, Hyunbae; Fu, Zhiyao; Qiu, Yining; Yang, Zhao; Wang, Jiemei; Zhang, Deqiang; Tong, Xin; Yin, Lei; Li, Jing; Wu, Jianmei; Qi, Nathan R.; Houten, Sander M.; Zhang, Ren

    2018-01-01

    The mitochondrial nicotinamide adenine dinucleotide (NAD) kinase (NADK2, also called MNADK) catalyzes phosphorylation of NAD to yield NADP. Little is known about the functions of mitochondrial NADP and MNADK in liver physiology and pathology. We investigated the effects of reduced mitochondrial NADP

  19. Properties of an NAD(H)-containing methanol dehydrogenase and its activator protein from Bacillus methanolicus

    NARCIS (Netherlands)

    Arfman, Nico; Hektor, Harm J.; Bystrykh, Leonid V.; Govorukhina, Natalya I.; Dijkhuizen, Lubbert; Frank, Johannes

    1997-01-01

    Oxidation of C1-C4 primary alcohols in thermotolerant Bacillus methanolicus strains is catalyzed by an NAD-dependent methanol dehydrogenase (MDH), composed of ten identical 43000-Mr subunits. Each MDH subunit contains a tightly, but non-covalently, bound NAD(H) molecule, in addition to 1 Zn2+ and

  20. Two NAD-linked redox shuttles maintain the peroxisomal redox balance in Saccharomyces cerevisiae

    NARCIS (Netherlands)

    Al-Saryi, Nadal A.; Al-Hejjaj, Murtakab Y.; van Roermund, Carlo W. T.; Hulmes, Georgia E.; Ekal, Lakhan; Payton, Chantell; Wanders, Ronald J. A.; Hettema, Ewald H.

    2017-01-01

    In Saccharomyces cerevisiae, peroxisomes are the sole site of fatty acid beta-oxidation. During this process, NAD(+) is reduced to NADH. When cells are grown on oleate medium, peroxisomal NADH is reoxidised to NAD(+) by malate dehydrogenase (Mdh3p) and reduction equivalents are transferred to the

  1. NADS: A Web Applet for Manipulation and Graphical Viewing of Nuclear Data

    International Nuclear Information System (INIS)

    McKinley, M S; Beck, B R; McNabb, D P

    2004-01-01

    We have developed a program called NADS (Nuclear and Atomic Data System) which provides a web-based, user-friendly interface for viewing nuclear data. NADS uses a client/server model. The client is a Java applet that runs in a web browser. The server is a Python code that delivers pointwise data to the applet per user request and then plots the data. The data is also stored in tables for viewing and modifying. NADS can display 2-D, 3-D and 4-D (time sliced) data in a powerful, user-friendly environment. Currently, evaluated nuclear data are available from ENDF/B-V, ENDF/B-VI, JENDL, JEF and Lawrence Livermore National Laboratory's ENDL databases. LLNL's ENDL database has data for neutron, gamma and charged particles as projectiles. In addition to displaying and saving data, NADS has the capability to perform computations with the data. NADS is accessible over the Internet at http://nuclear.llnl.gov/

  2. A conserved NAD+binding pocket that regulates protein-protein interactions during aging.

    Science.gov (United States)

    Li, Jun; Bonkowski, Michael S; Moniot, Sébastien; Zhang, Dapeng; Hubbard, Basil P; Ling, Alvin J Y; Rajman, Luis A; Qin, Bo; Lou, Zhenkun; Gorbunova, Vera; Aravind, L; Steegborn, Clemens; Sinclair, David A

    2017-03-24

    DNA repair is essential for life, yet its efficiency declines with age for reasons that are unclear. Numerous proteins possess Nudix homology domains (NHDs) that have no known function. We show that NHDs are NAD + (oxidized form of nicotinamide adenine dinucleotide) binding domains that regulate protein-protein interactions. The binding of NAD + to the NHD domain of DBC1 (deleted in breast cancer 1) prevents it from inhibiting PARP1 [poly(adenosine diphosphate-ribose) polymerase], a critical DNA repair protein. As mice age and NAD + concentrations decline, DBC1 is increasingly bound to PARP1, causing DNA damage to accumulate, a process rapidly reversed by restoring the abundance of NAD + Thus, NAD + directly regulates protein-protein interactions, the modulation of which may protect against cancer, radiation, and aging. Copyright © 2017, American Association for the Advancement of Science.

  3. The NAD(+) precursor nicotinamide riboside enhances oxidative metabolism and protects against high-fat diet-induced obesity

    NARCIS (Netherlands)

    Cantó, Carles; Houtkooper, Riekelt H.; Pirinen, Eija; Youn, Dou Y.; Oosterveer, Maaike H.; Cen, Yana; Fernandez-Marcos, Pablo J.; Yamamoto, Hiroyasu; Andreux, Pénélope A.; Cettour-Rose, Philippe; Gademann, Karl; Rinsch, Chris; Schoonjans, Kristina; Sauve, Anthony A.; Auwerx, Johan

    2012-01-01

    As NAD(+) is a rate-limiting cosubstrate for the sirtuin enzymes, its modulation is emerging as a valuable tool to regulate sirtuin function and, consequently, oxidative metabolism. In line with this premise, decreased activity of PARP-1 or CD38-both NAD(+) consumers-increases NAD(+)

  4. Phosphoribosyl diphosphate synthetase-independent NAD de novo synthesis in Escherichia coli: a new phenotype of phosphate regulon mutants

    DEFF Research Database (Denmark)

    Hove-Jensen, Bjarne

    1996-01-01

    reaction, an enzyme of the NAD de novo pathway. Several NAD-independent mutants of a host from which prs had been deleted were isolated; all of them were shown to have lesions in the pstSCAB-phoU operon, in which mutations lead to derepression of the Pho regulon. In addition NAD-independent growth...

  5. Sources and implications of NADH/NAD+ redox imbalance in diabetes and its complications

    Directory of Open Access Journals (Sweden)

    Wu J

    2016-05-01

    Full Text Available Jinzi Wu,1Zhen Jin,1Hong Zheng,1,2Liang-Jun Yan1 1Department of Pharmaceutical Sciences, UNT System College of Pharmacy, University of North Texas Health Science Center, Fort Worth, TX, USA; 2Department of Basic Theory of Traditional Chinese Medicine, College of Basic Medicine, Shandong University of Traditional Chinese Medicine, Jinan, People’s Republic of China Abstract: NAD+ is a fundamental molecule in metabolism and redox signaling. In diabetes and its complications, the balance between NADH and NAD+ can be severely perturbed. On one hand, NADH is overproduced due to influx of hyperglycemia to the glycolytic and Krebs cycle pathways and activation of the polyol pathway. On the other hand, NAD+ can be diminished or depleted by overactivation of poly ADP ribose polymerase that uses NAD+ as its substrate. Moreover, sirtuins, another class of enzymes that also use NAD+ as their substrate for catalyzing protein deacetylation reactions, can also affect cellular content of NAD+. Impairment of NAD+ regeneration enzymes such as lactate dehydrogenase in erythrocytes and complex I in mitochondria can also contribute to NADH accumulation and NAD+ deficiency. The consequence of NADH/NAD+ redox imbalance is initially reductive stress that eventually leads to oxidative stress and oxidative damage to macromolecules, including DNA, lipids, and proteins. Accordingly, redox imbalance-triggered oxidative damage has been thought to be a major factor contributing to the development of diabetes and its complications. Future studies on restoring NADH/NAD+ redox balance could provide further insights into design of novel antidiabetic strategies. Keywords: mitochondria, complex I, reactive oxygen species, polyol pathway, poly ADP ribosylation, sirtuins, oxidative stress, oxidative damage

  6. NAD(H) and NADP(H) Redox Couples and Cellular Energy Metabolism.

    Science.gov (United States)

    Xiao, Wusheng; Wang, Rui-Sheng; Handy, Diane E; Loscalzo, Joseph

    2018-01-20

    The nicotinamide adenine dinucleotide (NAD + )/reduced NAD + (NADH) and NADP + /reduced NADP + (NADPH) redox couples are essential for maintaining cellular redox homeostasis and for modulating numerous biological events, including cellular metabolism. Deficiency or imbalance of these two redox couples has been associated with many pathological disorders. Recent Advances: Newly identified biosynthetic enzymes and newly developed genetically encoded biosensors enable us to understand better how cells maintain compartmentalized NAD(H) and NADP(H) pools. The concept of redox stress (oxidative and reductive stress) reflected by changes in NAD(H)/NADP(H) has increasingly gained attention. The emerging roles of NAD + -consuming proteins in regulating cellular redox and metabolic homeostasis are active research topics. The biosynthesis and distribution of cellular NAD(H) and NADP(H) are highly compartmentalized. It is critical to understand how cells maintain the steady levels of these redox couple pools to ensure their normal functions and simultaneously avoid inducing redox stress. In addition, it is essential to understand how NAD(H)- and NADP(H)-utilizing enzymes interact with other signaling pathways, such as those regulated by hypoxia-inducible factor, to maintain cellular redox homeostasis and energy metabolism. Additional studies are needed to investigate the inter-relationships among compartmentalized NAD(H)/NADP(H) pools and how these two dinucleotide redox couples collaboratively regulate cellular redox states and cellular metabolism under normal and pathological conditions. Furthermore, recent studies suggest the utility of using pharmacological interventions or nutrient-based bioactive NAD + precursors as therapeutic interventions for metabolic diseases. Thus, a better understanding of the cellular functions of NAD(H) and NADP(H) may facilitate efforts to address a host of pathological disorders effectively. Antioxid. Redox Signal. 28, 251-272.

  7. The MiniBooNE detector

    International Nuclear Information System (INIS)

    Aguilar-Arevalo, A.A.; Anderson, C.E.; Bartoszek, L.M.; Bazarko, A.O.; Brice, S.J.; Brown, B.C.; Bugel, L.; Cao, J.; Coney, L.; Conrad, J.M.; Cox, D.C.; Curioni, A.; Djurcic, Z.; Finley, D.A.; Fleming, B.T.; Ford, R.; Garcia, F.G.; Garvey, G.T.; Green, C.; Green, J.A.

    2009-01-01

    The MiniBooNE neutrino detector was designed and built to look for ν μ →ν e oscillations in the (sin 2 2θ,Δm 2 ) parameter space region where the LSND experiment reported a signal. The MiniBooNE experiment used a beam energy and baseline that were an order of magnitude larger than those of LSND so that the backgrounds and systematic errors would be completely different. This paper provides a detailed description of the design, function, and performance of the MiniBooNE detector.

  8. Tributyltin induces mitochondrial fission through NAD-IDH dependent mitofusin degradation in human embryonic carcinoma cells.

    Science.gov (United States)

    Yamada, Shigeru; Kotake, Yaichiro; Nakano, Mizuho; Sekino, Yuko; Kanda, Yasunari

    2015-08-01

    Organotin compounds, such as tributyltin (TBT), are well-known endocrine disruptors. TBT acts at the nanomolar level through genomic pathways via the peroxisome proliferator activated receptor (PPAR)/retinoid X receptor (RXR). We recently reported that TBT inhibits cell growth and the ATP content in the human embryonic carcinoma cell line NT2/D1 via a non-genomic pathway involving NAD(+)-dependent isocitrate dehydrogenase (NAD-IDH), which metabolizes isocitrate to α-ketoglutarate. However, the molecular mechanisms by which NAD-IDH mediates TBT toxicity remain unclear. In the present study, we evaluated the effects of TBT on mitochondrial NAD-IDH and energy production. Staining with MitoTracker revealed that nanomolar TBT levels induced mitochondrial fragmentation. TBT also degraded the mitochondrial fusion proteins, mitofusins 1 and 2. Interestingly, apigenin, an inhibitor of NAD-IDH, mimicked the effects of TBT. Incubation with an α-ketoglutarate analogue partially recovered TBT-induced mitochondrial dysfunction, supporting the involvement of NAD-IDH. Our data suggest that nanomolar TBT levels impair mitochondrial quality control via NAD-IDH in NT2/D1 cells. Thus, mitochondrial function in embryonic cells could be used to assess cytotoxicity associated with metal exposure.

  9. Nicotinamidase participates in the salvage pathway of NAD biosynthesis in Arabidopsis.

    Science.gov (United States)

    Wang, Guodong; Pichersky, Eran

    2007-03-01

    Nicotinamide adenine dinucleotide (NAD) and nicotinamide adenine dinucleotide phosphate (NADP), which is derived from NAD, have important roles as a redox carriers in metabolism. A combination of de novo and salvage pathways contribute to the biosynthesis of NAD in all organisms. The pathways and enzymes of the NAD salvage pathway in yeast and animals, which diverge at nicotinamide, have been extensively studied. Yeast cells convert nicotinamide to nicotinic acid, while mammals lack the enzyme nicotinamidase and instead convert nicotinamide to nicotinamide mononucleotide. Here we show that Arabidopsis thaliana gene At2g22570 encodes a nicotinamidase, which is expressed in all tissues, with the highest levels observed in roots and stems. The 244-residue protein, designated AtNIC1, converts nicotinamide to nicotinic acid and has a Km value of 118 +/- 17 microM and a Kcat value of 0.93 +/- 0.13 sec(-1). Plants homozygous for a null AtNIC1 allele, nic1-1, have lower levels of NAD and NADP under normal growth conditions, indicating that AtNIC1 participates in a yeast-type NAD salvage pathway. Mutant plants also exhibit hypersensitivity to treatments of abscisic acid and NaCl, which is correlated with their inability to increase the cellular levels of NAD(H) under these growth conditions, as occurs in wild-type plants. We also show that the growth of the roots of wild-type but not nic1-1 mutant plants is inhibited and distorted by nicotinamide.

  10. Photolabeling of Glu-129 of the S-1 subunit of pertussis toxin with NAD

    Energy Technology Data Exchange (ETDEWEB)

    Barbieri, J.T.; Mende-Mueller, L.M.; Rappuoli, R.; Collier, R.J. (Medical College of Wisconsin, Milwaukee (USA))

    1989-11-01

    UV irradiation was shown to induce efficient transfer of radiolabel from nicotinamide-labeled NAD to a recombinant protein (C180 peptide) containing the catalytic region of the S-1 subunit of pertussis toxin. Incorporation of label from (3H-nicotinamide)NAD was efficient (0.5 to 0.6 mol/mol of protein) relative to incorporation from (32P-adenylate)NAD (0.2 mol/mol of protein). Label from (3H-nicotinamide)NAD was specifically associated with Glu-129. Replacement of Glu-129 with glycine or aspartic acid made the protein refractory to photolabeling with (3H-nicotinamide)NAD, whereas replacement of a nearby glutamic acid, Glu-139, with serine did not. Photolabeling of the C180 peptide with NAD is similar to that observed with diphtheria toxin and exotoxin A of Pseudomonas aeruginosa, in which the nicotinamide portion of NAD is transferred to Glu-148 and Glu-553, respectively, in the two toxins. These results implicate Glu-129 of the S-1 subunit as an active-site residue and a potentially important site for genetic modification of pertussis toxin for development of an acellular vaccine against Bordetella pertussis.

  11. Photolabeling of Glu-129 of the S-1 subunit of pertussis toxin with NAD

    International Nuclear Information System (INIS)

    Barbieri, J.T.; Mende-Mueller, L.M.; Rappuoli, R.; Collier, R.J.

    1989-01-01

    UV irradiation was shown to induce efficient transfer of radiolabel from nicotinamide-labeled NAD to a recombinant protein (C180 peptide) containing the catalytic region of the S-1 subunit of pertussis toxin. Incorporation of label from [3H-nicotinamide]NAD was efficient (0.5 to 0.6 mol/mol of protein) relative to incorporation from [32P-adenylate]NAD (0.2 mol/mol of protein). Label from [3H-nicotinamide]NAD was specifically associated with Glu-129. Replacement of Glu-129 with glycine or aspartic acid made the protein refractory to photolabeling with [3H-nicotinamide]NAD, whereas replacement of a nearby glutamic acid, Glu-139, with serine did not. Photolabeling of the C180 peptide with NAD is similar to that observed with diphtheria toxin and exotoxin A of Pseudomonas aeruginosa, in which the nicotinamide portion of NAD is transferred to Glu-148 and Glu-553, respectively, in the two toxins. These results implicate Glu-129 of the S-1 subunit as an active-site residue and a potentially important site for genetic modification of pertussis toxin for development of an acellular vaccine against Bordetella pertussis

  12. Ketone-Based Metabolic Therapy: Is Increased NAD+ a Primary Mechanism?

    Science.gov (United States)

    Elamin, Marwa; Ruskin, David N; Masino, Susan A; Sacchetti, Paola

    2017-01-01

    The ketogenic diet's (KD) anticonvulsant effects have been well-documented for nearly a century, including in randomized controlled trials. Some patients become seizure-free and some remain so after diet cessation. Many recent studies have explored its expanded therapeutic potential in diverse neurological disorders, yet no mechanism(s) of action have been established. The diet's high fat, low carbohydrate composition reduces glucose utilization and promotes the production of ketone bodies. Ketone bodies are a more efficient energy source than glucose and improve mitochondrial function and biogenesis. Cellular energy production depends on the metabolic coenzyme nicotinamide adenine dinucleotide (NAD), a marker for mitochondrial and cellular health. Furthermore, NAD activates downstream signaling pathways (such as the sirtuin enzymes) associated with major benefits such as longevity and reduced inflammation; thus, increasing NAD is a coveted therapeutic endpoint. Based on differential NAD + utilization during glucose- vs. ketone body-based acetyl-CoA generation for entry into the tricarboxylic cycle, we propose that a KD will increase the NAD + /NADH ratio. When rats were fed ad libitum KD, significant increases in hippocampal NAD + /NADH ratio and blood ketone bodies were detected already at 2 days and remained elevated at 3 weeks, indicating an early and persistent metabolic shift. Based on diverse published literature and these initial data we suggest that increased NAD during ketolytic metabolism may be a primary mechanism behind the beneficial effects of this metabolic therapy in a variety of brain disorders and in promoting health and longevity.

  13. Fumarate analogs act as allosteric inhibitors of the human mitochondrial NAD(P)+-dependent malic enzyme.

    Science.gov (United States)

    Hsieh, Ju-Yi; Liu, Jyung-Hurng; Yang, Pai-Chun; Lin, Chi-Li; Liu, Guang-Yaw; Hung, Hui-Chih

    2014-01-01

    Human mitochondrial NAD(P)+-dependent malic enzyme (m-NAD(P)-ME) is allosterically activated by the four-carbon trans dicarboxylic acid, fumarate. Previous studies have suggested that the dicarboxylic acid in a trans conformation around the carbon-carbon double bond is required for the allosteric activation of the enzyme. In this paper, the allosteric effects of fumarate analogs on m-NAD(P)-ME are investigated. Two fumarate-insensitive mutants, m-NAD(P)-ME_R67A/R91A and m-NAD(P)-ME_K57S/E59N/K73E/D102S, as well as c-NADP-ME, were used as the negative controls. Among these analogs, mesaconate, trans-aconitate, monomethyl fumarate and monoethyl fumarate were allosteric activators of the enzyme, while oxaloacetate, diethyl oxalacetate, and dimethyl fumarate were found to be allosteric inhibitors of human m-NAD(P)-ME. The IC50 value for diethyl oxalacetate was approximately 2.5 mM. This paper suggests that the allosteric inhibitors may impede the conformational change from open form to closed form and therefore inhibit m-NAD(P)-ME enzyme activity.

  14. Allosteric substrate inhibition of Arabidopsis NAD-dependent malic enzyme 1 is released by fumarate.

    Science.gov (United States)

    Tronconi, Marcos Ariel; Wheeler, Mariel Claudia Gerrard; Martinatto, Andrea; Zubimendi, Juan Pablo; Andreo, Carlos Santiago; Drincovich, María Fabiana

    2015-03-01

    Plant mitochondria can use L-malate and fumarate, which accumulate in large levels, as respiratory substrates. In part, this property is due to the presence of NAD-dependent malic enzymes (NAD-ME) with particular biochemical characteristics. Arabidopsis NAD-ME1 exhibits a non-hyperbolic behavior for the substrate L-malate, and its activity is strongly stimulated by fumarate. Here, the possible structural connection between these properties was explored through mutagenesis, kinetics, and fluorescence studies. The results indicated that NAD-ME1 has a regulatory site for L-malate that can also bind fumarate. L-Malate binding to this site elicits a sigmoidal and low substrate-affinity response, whereas fumarate binding turns NAD-ME1 into a hyperbolic and high substrate affinity enzyme. This effect was also observed when the allosteric site was either removed or altered. Hence, fumarate is not really an activator, but suppresses the inhibitory effect of l-malate. In addition, residues Arg50, Arg80 and Arg84 showed different roles in organic acid binding. These residues form a triad, which is the basis of the homo and heterotrophic effects that characterize NAD-ME1. The binding of L-malate and fumarate at the same allosteric site is herein reported for a malic enzyme and clearly indicates an important role of NAD-ME1 in processes that control flow of C4 organic acids in Arabidopsis mitochondrial metabolism. Copyright © 2014 Elsevier Ltd. All rights reserved.

  15. Fumarate analogs act as allosteric inhibitors of the human mitochondrial NAD(P+-dependent malic enzyme.

    Directory of Open Access Journals (Sweden)

    Ju-Yi Hsieh

    Full Text Available Human mitochondrial NAD(P+-dependent malic enzyme (m-NAD(P-ME is allosterically activated by the four-carbon trans dicarboxylic acid, fumarate. Previous studies have suggested that the dicarboxylic acid in a trans conformation around the carbon-carbon double bond is required for the allosteric activation of the enzyme. In this paper, the allosteric effects of fumarate analogs on m-NAD(P-ME are investigated. Two fumarate-insensitive mutants, m-NAD(P-ME_R67A/R91A and m-NAD(P-ME_K57S/E59N/K73E/D102S, as well as c-NADP-ME, were used as the negative controls. Among these analogs, mesaconate, trans-aconitate, monomethyl fumarate and monoethyl fumarate were allosteric activators of the enzyme, while oxaloacetate, diethyl oxalacetate, and dimethyl fumarate were found to be allosteric inhibitors of human m-NAD(P-ME. The IC50 value for diethyl oxalacetate was approximately 2.5 mM. This paper suggests that the allosteric inhibitors may impede the conformational change from open form to closed form and therefore inhibit m-NAD(P-ME enzyme activity.

  16. Muscle type-specific responses to NAD+ salvage biosynthesis promote muscle function in Caenorhabditis elegans.

    Science.gov (United States)

    Vrablik, Tracy L; Wang, Wenqing; Upadhyay, Awani; Hanna-Rose, Wendy

    2011-01-15

    Salvage biosynthesis of nicotinamide adenine dinucleotide (NAD(+)) from nicotinamide (NAM) lowers NAM levels and replenishes the critical molecule NAD(+) after it is hydrolyzed. This pathway is emerging as a regulator of multiple biological processes. Here we probe the contribution of the NAM-NAD(+) salvage pathway to muscle development and function using Caenorhabditis elegans. C. elegans males with mutations in the nicotinamidase pnc-1, which catalyzes the first step of this NAD(+) salvage pathway, cannot mate due to a spicule muscle defect. Multiple muscle types are impaired in the hermaphrodites, including body wall muscles, pharyngeal muscles and vulval muscles. An active NAD(+) salvage pathway is required for optimal function of each muscle cell type. However, we found surprising muscle-cell-type specificity in terms of both the timing and relative sensitivity to perturbation of NAD(+) production or NAM levels. Active NAD(+) biosynthesis during development is critical for function of the male spicule protractor muscles during adulthood, but these muscles can surprisingly do without salvage biosynthesis in adulthood under the conditions examined. The body wall muscles require ongoing NAD(+) salvage biosynthesis both during development and adulthood for maximum function. The vulval muscles do not function in the presence of elevated NAM concentrations, but NAM supplementation is only slightly deleterious to body wall muscles during development or upon acute application in adults. Thus, the pathway plays distinct roles in different tissues. As NAM-NAD(+) biosynthesis also impacts muscle differentiation in vertebrates, we propose that similar complexities may be found among vertebrate muscle cell types. Copyright © 2010 Elsevier Inc. All rights reserved.

  17. Alteration in substrate specificity of horse liver alcohol dehydrogenase by an acyclic nicotinamide analog of NAD(+).

    Science.gov (United States)

    Malver, Olaf; Sebastian, Mina J; Oppenheimer, Norman J

    2014-11-01

    A new, acyclic NAD-analog, acycloNAD(+) has been synthesized where the nicotinamide ribosyl moiety has been replaced by the nicotinamide (2-hydroxyethoxy)methyl moiety. The chemical properties of this analog are comparable to those of β-NAD(+) with a redox potential of -324mV and a 341nm λmax for the reduced form. Both yeast alcohol dehydrogenase (YADH) and horse liver alcohol dehydrogenase (HLADH) catalyze the reduction of acycloNAD(+) by primary alcohols. With HLADH 1-butanol has the highest Vmax at 49% that of β-NAD(+). The primary deuterium kinetic isotope effect is greater than 3 indicating a significant contribution to the rate limiting step from cleavage of the carbon-hydrogen bond. The stereochemistry of the hydride transfer in the oxidation of stereospecifically deuterium labeled n-butanol is identical to that for the reaction with β-NAD(+). In contrast to the activity toward primary alcohols there is no detectable reduction of acycloNAD(+) by secondary alcohols with HLADH although these alcohols serve as competitive inhibitors. The net effect is that acycloNAD(+) has converted horse liver ADH from a broad spectrum alcohol dehydrogenase, capable of utilizing either primary or secondary alcohols, into an exclusively primary alcohol dehydrogenase. This is the first example of an NAD analog that alters the substrate specificity of a dehydrogenase and, like site-directed mutagenesis of proteins, establishes that modifications of the coenzyme distance from the active site can be used to alter enzyme function and substrate specificity. These and other results, including the activity with α-NADH, clearly demonstrate the promiscuity of the binding interactions between dehydrogenases and the riboside phosphate of the nicotinamide moiety, thus greatly expanding the possibilities for the design of analogs and inhibitors of specific dehydrogenases. Copyright © 2014 Elsevier B.V. All rights reserved.

  18. Web life: Mahalo.ne.Trash

    Science.gov (United States)

    2014-05-01

    Mahalo.ne.Trash is the personal blog of John Asher Johnson, an astronomer at Harvard University in the US who began blogging in 2007, when he was about to begin a postdoctoral fellowship at the University of Hawaii.

  19. UCB-NE-107 user's manual

    International Nuclear Information System (INIS)

    Lee, W.W.L.

    1989-03-01

    The purpose of this manual is to provide users of UCB-NE-107 with the information necessary to use UCB-NE-107 effectively. UCB-NE-107 is a computer code for calculating the fractional rate of readily soluble radionuclides that are released from nuclear waste emplaced in water-saturated porous media. Waste placed in such environments will gradually dissolve. For many species such as actinides and rare earths, the process of dissolution is governed by the exterior flow field, and the chemical reaction rate or leaching rate. However, for readily soluble species such as 135 Cs, 137 Cs, and 129 I, it has been observed that their dissolution rates are rapid. UCB-NE-107 is a code for calculating the release rate at the waste/rock interface, to check compliance with the US Nuclear Regulatory Commission's (USNRC) subsystem performance objective. It is an implementation of the analytic solution given below. 5 refs., 2 figs

  20. Is any awareness necessary for an Ne?

    Directory of Open Access Journals (Sweden)

    Shani eShalgi

    2012-05-01

    Full Text Available The Error-Related Negativity (Ne or ERN is a reliable electrophysiological index of error processing, which has been found to be independent of whether a subject is aware of an error or not. A large Ne was equally seen after errors that were consciously detected (Aware errors and those that were not (Unaware errors, compared to a small negativity for correct responses (CRN. This suggests a dissociation between an automatic, preconscious error processing mechanism and subjective evaluation. A common concern regarding this finding is that subjects could have been somewhat aware of their errors, but did not report them due to lack of confidence. Here we tested this possibility directly using a betting paradigm which allowed us to separate occasions in which the subjects were confident of their response and trials in which they were unsure. In a choice reaction time task, subjects directly judged the accuracy of each response (correct or error and then bet on this judgment using a high, medium or low amount of money. The bets were used to determine the level of confidence the subjects had of their response. The average across all subjects regardless of confidence (betting measure replicated the reported finding of an equal Ne for Aware and Unaware errors which was larger than the CRN. However, when Ne measurement was confined to high confidence (high bet trials in confident subjects, a prominent Ne was seen only for Aware errors, while confident Unaware errors (i.e., error trials on which subjects made high bets that they were correct elicited a response that did not differ from the CRN elicited by truly correct answers. In contrast, for low confidence trials in unconfident subjects, an intermediate and equal Ne/CRN was elicited by Correct responses, Aware and Unaware errors. These results provide direct evidence that the Ne is related to error awareness, and suggest the amplitude of the Ne/CRN depends on individual differences in error reporting

  1. Nicotinic acid, nicotinamide, and nicotinamide riboside: a molecular evaluation of NAD+ precursor vitamins in human nutrition.

    Science.gov (United States)

    Bogan, Katrina L; Brenner, Charles

    2008-01-01

    Although baseline requirements for nicotinamide adenine dinucleotide (NAD+) synthesis can be met either with dietary tryptophan or with less than 20 mg of daily niacin, which consists of nicotinic acid and/or nicotinamide, there is growing evidence that substantially greater rates of NAD+ synthesis may be beneficial to protect against neurological degeneration, Candida glabrata infection, and possibly to enhance reverse cholesterol transport. The distinct and tissue-specific biosynthetic and/or ligand activities of tryptophan, nicotinic acid, nicotinamide, and the newly identified NAD+ precursor, nicotinamide riboside, reviewed herein, are responsible for vitamin-specific effects and side effects. Because current data suggest that nicotinamide riboside may be the only vitamin precursor that supports neuronal NAD+ synthesis, we present prospects for human nicotinamide riboside supplementation and propose areas for future research.

  2. Mitochondrial type II NAD(PH dehydrogenases in fungal cell death

    Directory of Open Access Journals (Sweden)

    A. Pedro Gonçalves

    2015-03-01

    Full Text Available During aerobic respiration, cells produce energy through oxidative phosphorylation, which includes a specialized group of multi-subunit complexes in the inner mitochondrial membrane known as the electron transport chain. However, this canonical pathway is branched into single polypeptide alternative routes in some fungi, plants, protists and bacteria. They confer metabolic plasticity, allowing cells to adapt to different environmental conditions and stresses. Type II NAD(PH dehydrogenases (also called alternative NAD(PH dehydrogenases are non-proton pumping enzymes that bypass complex I. Recent evidence points to the involvement of fungal alternative NAD(PH dehydrogenases in the process of programmed cell death, in addition to their action as overflow systems upon oxidative stress. Consistent with this, alternative NAD(PH dehydrogenases are phylogenetically related to cell death - promoting proteins of the apoptosis-inducing factor (AIF-family.

  3. US Coast Guard Stations in Louisiana, Geographic NAD83, USCG [coast_guard_stations_USCG_1997

    Data.gov (United States)

    Louisiana Geographic Information Center — This is is a point dataset for the locations and attributes of eight US Coast Guard stations in Louisiana. The attributes include name, address, latitude (NAD27),...

  4. Mutations that Allow SIR2 Orthologs to Function in a NAD+-Depleted Environment

    Directory of Open Access Journals (Sweden)

    Caitlin R. Ondracek

    2017-03-01

    Full Text Available Sirtuin enzymes depend on NAD+ to catalyze protein deacetylation. Therefore, the lowering of NAD+ during aging leads to decreased sirtuin activity and may speed up aging processes in laboratory animals and humans. In this study, we used a genetic screen to identify two mutations in the catalytic domain of yeast Sir2 that allow the enzyme to function in an NAD+-depleted environment. These mutant enzymes give rise to a significant increase of yeast replicative lifespan and increase deacetylation by the Sir2 ortholog, SIRT1, in mammalian cells. Our data suggest that these mutations increase the stability of the conserved catalytic sirtuin domain, thereby increasing the catalytic efficiency of the mutant enzymes. Our approach to identifying sirtuin mutants that permit function in NAD+-limited environments may inform the design of small molecules that can maintain sirtuin activity in aging organisms.

  5. Vibrio Phage KVP40 Encodes a Functional NAD+Salvage Pathway.

    Science.gov (United States)

    Lee, Jae Yun; Li, Zhiqun; Miller, Eric S

    2017-05-01

    The genome of T4-type Vibrio bacteriophage KVP40 has five genes predicted to encode proteins of pyridine nucleotide metabolism, of which two, nadV and natV , would suffice for an NAD + salvage pathway. NadV is an apparent nicotinamide phosphoribosyltransferase (NAmPRTase), and NatV is an apparent bifunctional nicotinamide mononucleotide adenylyltransferase (NMNATase) and nicotinamide-adenine dinucleotide pyrophosphatase (Nudix hydrolase). Genes encoding the predicted salvage pathway were cloned and expressed in Escherichia coli , the proteins were purified, and their enzymatic properties were examined. KVP40 NadV NAmPRTase is active in vitro , and a clone complements a Salmonella mutant defective in both the bacterial de novo and salvage pathways. Similar to other NAmPRTases, the KVP40 enzyme displayed ATPase activity indicative of energy coupling in the reaction mechanism. The NatV NMNATase activity was measured in a coupled reaction system demonstrating NAD + biosynthesis from nicotinamide, phosphoribosyl pyrophosphate, and ATP. The NatV Nudix hydrolase domain was also shown to be active, with preferred substrates of ADP-ribose, NAD + , and NADH. Expression analysis using reverse transcription-quantitative PCR (qRT-PCR) and enzyme assays of infected Vibrio parahaemolyticus cells demonstrated nadV and natV transcription during the early and delayed-early periods of infection when other KVP40 genes of nucleotide precursor metabolism are expressed. The distribution and phylogeny of NadV and NatV proteins among several large double-stranded DNA (dsDNA) myophages, and also those from some very large siphophages, suggest broad relevance of pyridine nucleotide scavenging in virus-infected cells. NAD + biosynthesis presents another important metabolic resource control point by large, rapidly replicating dsDNA bacteriophages. IMPORTANCE T4-type bacteriophages enhance DNA precursor synthesis through reductive reactions that use NADH/NADPH as the electron donor and NAD

  6. Isotope partitioning for NAD-malic enzyme from Ascaris suum confirms a steady-state random kinetic mechanism

    Energy Technology Data Exchange (ETDEWEB)

    Chen, C.Y.; Harris, B.G.; Cook, P.F.

    1988-01-12

    Isotope partitioning studies beginning with E-(/sup 14/C)NAD, E-(/sup 14/C) malate, E-(/sup 14/C) NAD-Mg/sup 2 +/, and E-Mg-(/sup 14/C)malate suggest a steady-state random mechanism for the NAD-malic enzyme. Isotope trapping beginning with E-(/sup 14/C)NAD and with varying concentrations of Mg/sup 2 +/ and malate in the chase solution indicates that Mg/sup 2 +/ is added in rapid equilibrium and must be added prior to malate for productive ternary complex formation. Equal percentage trapping from E-(/sup 14/C)NAD-Mg and E-Mg-(/sup 14/C) malate indicates the mechanism is steady-state random with equal off-rates for NAD and malate from E-NAD-Mg-malate. The off-rates for both do not change significantly in the ternary E-Mg-malate and E-NAD-Mg complexes, nor does the off-rate change for NAD from E-NAD. No trapping of malate was obtained from E-(/sup 14/C) malate, suggesting that this complex is nonproductive. A quantitative analysis of the data allows an estimation of values for a number of the rate constants along the reaction pathway.

  7. Isotope partitioning for NAD-malic enzyme from Ascaris suum confirms a steady-state random kinetic mechanism

    International Nuclear Information System (INIS)

    Chen, C.Y.; Harris, B.G.; Cook, P.F.

    1988-01-01

    Isotope partitioning studies beginning with E-[ 14 C]NAD, E-[ 14 C] malate, E-[ 14 C] NAD-Mg 2+ , and E-Mg-[ 14 C]malate suggest a steady-state random mechanism for the NAD-malic enzyme. Isotope trapping beginning with E-[ 14 C]NAD and with varying concentrations of Mg 2+ and malate in the chase solution indicates that Mg 2+ is added in rapid equilibrium and must be added prior to malate for productive ternary complex formation. Equal percentage trapping from E-[ 14 C]NAD-Mg and E-Mg-[ 14 C] malate indicates the mechanism is steady-state random with equal off-rates for NAD and malate from E-NAD-Mg-malate. The off-rates for both do not change significantly in the ternary E-Mg-malate and E-NAD-Mg complexes, nor does the off-rate change for NAD from E-NAD. No trapping of malate was obtained from E-[ 14 C] malate, suggesting that this complex is nonproductive. A quantitative analysis of the data allows an estimation of values for a number of the rate constants along the reaction pathway

  8. MicroBooNE: The Search For The MiniBooNE Low Energy Excess

    Energy Technology Data Exchange (ETDEWEB)

    Kaleko, David [Columbia Univ., New York, NY (United States)

    2017-01-01

    This thesis describes work towards the search for a low energy excess in MicroBooNE. What MicroBooNE is, what the low energy excess is, and how one searches for the latter in the former will be described in detail.

  9. Faktor-faktor Yang Berhubungan Dengan Kejadian Underweight Pada Anak Usia 24-59 Bulan Di Nanggroe Aceh Darussalam (Nad): Analisis Data Surkesda Nad 2006

    OpenAIRE

    Mulyati, Sri; Sandjaja, Sandjaja; Tjandrarini, Dwi Hapsari

    2008-01-01

    Determinant Factors of The Incidence of Underweight Children Ageg 24-59 Month In Nanggroe Aceh Darussalam (NAD).Background: Surkesda NAD 2006 as post tsunami household health survey and covering all 21 districts/cities had assessed child nutritional status (underweight) for age 24-59 months, with cut-off point <-2.00 SD. A total sample of 1,437 children aged 0-59 months was included in the assessment.Objectives: The objective of this study is to analyze child nutritional status and factors...

  10. NAD-dependent isocitrate dehydrogenase as a novel target of tributyltin in human embryonic carcinoma cells

    Science.gov (United States)

    Yamada, Shigeru; Kotake, Yaichiro; Demizu, Yosuke; Kurihara, Masaaki; Sekino, Yuko; Kanda, Yasunari

    2014-08-01

    Tributyltin (TBT) is known to cause developmental defects as endocrine disruptive chemicals (EDCs). At nanomoler concentrations, TBT actions were mediated by genomic pathways via PPAR/RXR. However, non-genomic target of TBT has not been elucidated. To investigate non-genomic TBT targets, we performed comprehensive metabolomic analyses using human embryonic carcinoma NT2/D1 cells. We found that 100 nM TBT reduced the amounts of α-ketoglutarate, succinate and malate. We further found that TBT decreased the activity of NAD-dependent isocitrate dehydrogenase (NAD-IDH), which catalyzes the conversion of isocitrate to α-ketoglutarate in the TCA cycle. In addition, TBT inhibited cell growth and enhanced neuronal differentiation through NAD-IDH inhibition. Furthermore, studies using bacterially expressed human NAD-IDH and in silico simulations suggest that TBT inhibits NAD-IDH due to a possible interaction. These results suggest that NAD-IDH is a novel non-genomic target of TBT at nanomolar levels. Thus, a metabolomic approach may provide new insights into the mechanism of EDC action.

  11. A Regulatory Role of NAD Redox Status on Flavin Cofactor Homeostasis in S. cerevisiae Mitochondria

    Directory of Open Access Journals (Sweden)

    Teresa Anna Giancaspero

    2013-01-01

    Full Text Available Flavin adenine dinucleotide (FAD and nicotinamide adenine dinucleotide (NAD are two redox cofactors of pivotal importance for mitochondrial functionality and cellular redox balance. Despite their relevance, the mechanism by which intramitochondrial NAD(H and FAD levels are maintained remains quite unclear in Saccharomyces cerevisiae. We investigated here the ability of isolated mitochondria to degrade externally added FAD and NAD (in both its reduced and oxidized forms. A set of kinetic experiments demonstrated that mitochondrial FAD and NAD(H destroying enzymes are different from each other and from the already characterized NUDIX hydrolases. We studied here, in some detail, FAD pyrophosphatase (EC 3.6.1.18, which is inhibited by NAD+ and NADH according to a noncompetitive inhibition, with Ki values that differ from each other by an order of magnitude. These findings, together with the ability of mitochondrial FAD pyrophosphatase to metabolize endogenous FAD, presumably deriving from mitochondrial holoflavoproteins destined to degradation, allow for proposing a novel possible role of mitochondrial NAD redox status in regulating FAD homeostasis and/or flavoprotein degradation in S. cerevisiae.

  12. Crystal structure of NAD+-dependent Peptoniphilus asaccharolyticus glutamate dehydrogenase reveals determinants of cofactor specificity.

    Science.gov (United States)

    Oliveira, Tânia; Panjikar, Santosh; Carrigan, John B; Hamza, Muaawia; Sharkey, Michael A; Engel, Paul C; Khan, Amir R

    2012-02-01

    Glutamate dehydrogenases (EC 1.4.1.2-4) catalyse the oxidative deamination of l-glutamate to α-ketoglutarate using NAD(P) as a cofactor. The bacterial enzymes are hexamers and each polypeptide consists of an N-terminal substrate-binding (Domain I) followed by a C-terminal cofactor-binding segment (Domain II). The reaction takes place at the junction of the two domains, which move as rigid bodies and are presumed to narrow the cleft during catalysis. Distinct signature sequences in the nucleotide-binding domain have been linked to NAD(+) vs. NADP(+) specificity, but they are not unambiguous predictors of cofactor preferences. Here, we have determined the crystal structure of NAD(+)-specific Peptoniphilus asaccharolyticus glutamate dehydrogenase in the apo state. The poor quality of native crystals was resolved by derivatization with selenomethionine, and the structure was solved by single-wavelength anomalous diffraction methods. The structure reveals an open catalytic cleft in the absence of substrate and cofactor. Modeling of NAD(+) in Domain II suggests that a hydrophobic pocket and polar residues contribute to nucleotide specificity. Mutagenesis and isothermal titration calorimetry studies of a critical glutamate at the P7 position of the core fingerprint confirms its role in NAD(+) binding. Finally, the cofactor binding site is compared with bacterial and mammalian enzymes to understand how the amino acid sequences and three-dimensional structures may distinguish between NAD(+) vs. NADP(+) recognition. Copyright © 2011 Elsevier Inc. All rights reserved.

  13. MacroH2A1.1 regulates mitochondrial respiration by limiting nuclear NAD+ consumption.

    Science.gov (United States)

    Posavec Marjanović, Melanija; Hurtado-Bagès, Sarah; Lassi, Maximilian; Valero, Vanesa; Malinverni, Roberto; Delage, Hélène; Navarro, Miriam; Corujo, David; Guberovic, Iva; Douet, Julien; Gama-Perez, Pau; Garcia-Roves, Pablo M; Ahel, Ivan; Ladurner, Andreas G; Yanes, Oscar; Bouvet, Philippe; Suelves, Mònica; Teperino, Raffaele; Pospisilik, J Andrew; Buschbeck, Marcus

    2017-11-01

    Histone variants are structural components of eukaryotic chromatin that can replace replication-coupled histones in the nucleosome. The histone variant macroH2A1.1 contains a macrodomain capable of binding NAD + -derived metabolites. Here we report that macroH2A1.1 is rapidly induced during myogenic differentiation through a switch in alternative splicing, and that myotubes that lack macroH2A1.1 have a defect in mitochondrial respiratory capacity. We found that the metabolite-binding macrodomain was essential for sustained optimal mitochondrial function but dispensable for gene regulation. Through direct binding, macroH2A1.1 inhibits basal poly-ADP ribose polymerase 1 (PARP-1) activity and thus reduces nuclear NAD + consumption. The resultant accumulation of the NAD + precursor NMN allows for maintenance of mitochondrial NAD + pools that are critical for respiration. Our data indicate that macroH2A1.1-containing chromatin regulates mitochondrial respiration by limiting nuclear NAD + consumption and establishing a buffer of NAD + precursors in differentiated cells.

  14. Involvement of cytosolic NAD+ glycohydrolase in cyclic ADP-ribose metabolism.

    Science.gov (United States)

    Matsumura, N; Tanuma, S

    1998-12-18

    The NAD+ glycohydrolase homogeneously purified from bovine brain cytosol was found to catalyze the synthesis and hydrolysis of cyclic ADP-ribose (cADPR). Although the formation of cADPR from NAD+ does not exceed about 2% of the reaction products, the cyclase activity is clearly evidenced by its conversion of NGD+ to cyclic GDP-ribose (cGDPR), which cannot be hydrolyzed to GDPR. Importantly, a steep increase in cADPR hydrolytic activity was observed at cADPR concentrations above 60 microM, which could be reproduced on a Hill curve with a Hill coefficient of 2. Thus, the allosteric binding of cADPR to the NAD+ glycohydrolase (E) molecule promotes the hydrolysis of cADPR. These results suggest that NAD+ hydrolysis to ADPR and nicotinamide catalyzed by the NAD+ glycohydrolase occurs through the formation of a cADPR. E. cADP-ribosyl complex. The low production of cADPR by NAD+ glycohydrolase compared with invertebrate ADP-ribosyl cyclase is believed to be attributable to the fast hydrolysis of cADPR by the allosteric effect of cADPR bound to the same enzyme that produces it. Copyright 1998 Academic Press.

  15. Reaction cross section for Ne isotopes

    International Nuclear Information System (INIS)

    Panda, R.N.; Sahu, B.K.; Patra, S.K.

    2012-01-01

    In the present contribution, first the bulk properties are calculated, such as binding energy (BE), root mean square charge radius r ch , matter radius r m and quadrupole deformation parameter β 2 for 18-32 Ne isotopes in the Relativistic mean field (RMF) and effective field theory motivated RMF (E-RMF) formalisms . Then the total nuclear reaction cross section σR is analyzes for the scattering of 20 Ne and 28-32 Ne from a 12 C target at 240 MeV/nucleon by using the RMF model. Thus the objective of the present study is to calculate the bulk properties as well as a systematic analysis of σR over a range of neutron rich nuclei in the frame work of Glauber model

  16. MicroBooNE: The Search For The MiniBooNE Low Energy Excess

    Science.gov (United States)

    Kaleko, David

    This thesis describes work towards the search for a low energy excess of electromagnetic events in the MicroBooNE detector. A background primer on the current state of neutrino physics is provided, including a description of the MiniBooNE detector and its published observation of an excess of electromagnetic events at low energies. A description of the MicroBooNE Liquid Argon Time Projection Chamber (LArTPC) detector is given, along with a description of the event selection and reconstruction algorithms developed to select electron neutrino charge-current interactions. A MiniBooNE-like signal is simulated in MicroBooNE with assumptions about the origin of the excess, and the sensitivity to observe such a signal above backgrounds in MicroBooNE is computed. An additional analysis is presented which constrains a dominant background in the MicroBooNE low energy excess search: the beam-intrinsic electron neutrino interactions which come from kaon decay in the beam-line. An essential step in this analysis is to reconstruct the energy of muon neutrino charge-current interactions in which the muon produced in the interaction escapes the detector. A publication detailing the algorithm which leverages the phenomenon of multiple Coulomb scattering to reconstruct the energy of escaping muons is provided as an appendix.

  17. A bionanohybrid ZnAl-NADS ecological pesticide as a treatment for soft rot disease in potato (Solanum tuberosum L.).

    Science.gov (United States)

    Morales-Irigoyen, Erika Elizabeth; de Las Mercedes Gómez-Y-Gómez, Yolanda; Flores-Moreno, Jorge Luis; Franco-Hernández, Marina Olivia

    2017-09-18

    Pectobacterium carotovorum (Pc) is a phytopathogenic strain that causes soft rot disease in potato (Solanum tuberosum L.), resulting in postharvest losses. Chemical control is effective for managing this disease, but overdoses cause adverse effects. Because farmers insist on using chemical agents for crop protection, it is necessary to develop more effective pesticides in which the active compound released can be regulated. In this context, we proposed the synthesis of ZnAl-NADS, in which nalidixic acid sodium salt (NADS) is linked to a ZnAl-NO 3 layered double hydroxide (LDH) host as a nanocarrier. XRD, FT-IR, and SEM analyses confirmed the successful intercalation of NADS into the interplanar LDH space. The drug release profile indicated that the maximum release was completed in 70 or 170 min for free NADS (alone) or for NADS released from ZnAl-NADS, respectively. This slow release was attributed to strong electrostatic interactions between the drug and the anion exchanger. A modulated release is preferable to the action of the bulk NADS, showing increased effectiveness and minimizing the amount of the chemical available to pollute the soil and the water. The fitting data from modified Freundlich and parabolic diffusion models explain the release behavior of the NADS, suggesting that the drug released from ZnAl-NADS bionanohybrid was carried out from the interlamellar sites, according to the ion exchange diffusion process also involving intraparticle diffusion (coeffect). ZnAl-NADS was tested in vitro against Escherichia coli (Ec) and Pc and exhibited bacteriostatic and biocidal effects at 0.025 and 0.075 mg mL -1 , respectively. ZnAl-NADS was also tested in vivo as an ecological pesticide for combating potato soft rot and was found to delay typical disease symptoms. In conclusion, ZnAl-NADS can potentially be used to control pests, infestation, and plant disease.

  18. Insight into molecular and functional properties of NMNAT3 reveals new hints of NAD homeostasis within human mitochondria

    OpenAIRE

    Felici, R; Lapucci, A; Ramazzotti, M; Chiarugi, A.

    2013-01-01

    Among the enzymes involved in NAD homeostasis, nicotinamide mononucleotide adenylyltransferases (NMNAT1-3) are central to intracellular NAD formation. Although NMNAT3 is postulated to be a mitochondrial enzyme contributing to NAD-dependent organelle functioning, information on endogenous proteins is lacking. We report that in human cells a single gene nmnat3 localized on chromosome 3 codes for two mRNA splice variants NMNATv1 and FKSG76, whereas the previously reported NMNAT3v2 transcript is ...

  19. The NAD+ metabolism of Leishmania, notably the enzyme nicotinamidase involved in NAD+ salvage, offers prospects for development of anti-parasite chemotherapy.

    Science.gov (United States)

    Michels, Paul A M; Avilán, Luisana

    2011-10-01

    NAD+ plays multiple, essential roles in the cell. As a cofactor in many redox reactions it is key in the cellular energy metabolism and as a substrate it participates in many reactions leading to a variety of covalent modifications of enzymes with major roles in regulation of expression and metabolism. Cells may have the ability to produce this metabolite either via alternative de novo synthesis pathways and/or by different salvage pathways. In this issue of Molecular Microbiology, Gazanion et al. (2011) demonstrate that Leishmania species can only rely on the salvage of NAD+ building blocks. One of the enzymes involved, nicotinamidase, is absent from human cells. The enzyme is important for growth of Leishmania infantum and essential for establishing an infection. The crystal structure of the parasite protein has been solved and shows prospects for design of inhibitors to be used as leads for development of new drugs. Indeed, NAD+ metabolism is currently being considered as a promising drug target in various diseases and the vulnerability of Leishmania for interference of this metabolism has been proved in previous work by the same group, by showing that administration of NAD+ precursors has detrimental effect on the pathogenic, amastigote stage of this parasite. © 2011 Blackwell Publishing Ltd.

  20. Generation, Release, and Uptake of the NAD Precursor Nicotinic Acid Riboside by Human Cells*

    Science.gov (United States)

    Kulikova, Veronika; Shabalin, Konstantin; Nerinovski, Kirill; Dölle, Christian; Niere, Marc; Yakimov, Alexander; Redpath, Philip; Khodorkovskiy, Mikhail; Migaud, Marie E.; Ziegler, Mathias; Nikiforov, Andrey

    2015-01-01

    NAD is essential for cellular metabolism and has a key role in various signaling pathways in human cells. To ensure proper control of vital reactions, NAD must be permanently resynthesized. Nicotinamide and nicotinic acid as well as nicotinamide riboside (NR) and nicotinic acid riboside (NAR) are the major precursors for NAD biosynthesis in humans. In this study, we explored whether the ribosides NR and NAR can be generated in human cells. We demonstrate that purified, recombinant human cytosolic 5′-nucleotidases (5′-NTs) CN-II and CN-III, but not CN-IA, can dephosphorylate the mononucleotides nicotinamide mononucleotide and nicotinic acid mononucleotide (NAMN) and thus catalyze NR and NAR formation in vitro. Similar to their counterpart from yeast, Sdt1, the human 5′-NTs require high (millimolar) concentrations of nicotinamide mononucleotide or NAMN for efficient catalysis. Overexpression of FLAG-tagged CN-II and CN-III in HEK293 and HepG2 cells resulted in the formation and release of NAR. However, NAR accumulation in the culture medium of these cells was only detectable under conditions that led to increased NAMN production from nicotinic acid. The amount of NAR released from cells engineered for increased NAMN production was sufficient to maintain viability of surrounding cells unable to use any other NAD precursor. Moreover, we found that untransfected HeLa cells produce and release sufficient amounts of NAR and NR under normal culture conditions. Collectively, our results indicate that cytosolic 5′-NTs participate in the conversion of NAD precursors and establish NR and NAR as integral constituents of human NAD metabolism. In addition, they point to the possibility that different cell types might facilitate each other's NAD supply by providing alternative precursors. PMID:26385918

  1. Generation, Release, and Uptake of the NAD Precursor Nicotinic Acid Riboside by Human Cells.

    Science.gov (United States)

    Kulikova, Veronika; Shabalin, Konstantin; Nerinovski, Kirill; Dölle, Christian; Niere, Marc; Yakimov, Alexander; Redpath, Philip; Khodorkovskiy, Mikhail; Migaud, Marie E; Ziegler, Mathias; Nikiforov, Andrey

    2015-11-06

    NAD is essential for cellular metabolism and has a key role in various signaling pathways in human cells. To ensure proper control of vital reactions, NAD must be permanently resynthesized. Nicotinamide and nicotinic acid as well as nicotinamide riboside (NR) and nicotinic acid riboside (NAR) are the major precursors for NAD biosynthesis in humans. In this study, we explored whether the ribosides NR and NAR can be generated in human cells. We demonstrate that purified, recombinant human cytosolic 5'-nucleotidases (5'-NTs) CN-II and CN-III, but not CN-IA, can dephosphorylate the mononucleotides nicotinamide mononucleotide and nicotinic acid mononucleotide (NAMN) and thus catalyze NR and NAR formation in vitro. Similar to their counterpart from yeast, Sdt1, the human 5'-NTs require high (millimolar) concentrations of nicotinamide mononucleotide or NAMN for efficient catalysis. Overexpression of FLAG-tagged CN-II and CN-III in HEK293 and HepG2 cells resulted in the formation and release of NAR. However, NAR accumulation in the culture medium of these cells was only detectable under conditions that led to increased NAMN production from nicotinic acid. The amount of NAR released from cells engineered for increased NAMN production was sufficient to maintain viability of surrounding cells unable to use any other NAD precursor. Moreover, we found that untransfected HeLa cells produce and release sufficient amounts of NAR and NR under normal culture conditions. Collectively, our results indicate that cytosolic 5'-NTs participate in the conversion of NAD precursors and establish NR and NAR as integral constituents of human NAD metabolism. In addition, they point to the possibility that different cell types might facilitate each other's NAD supply by providing alternative precursors. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

  2. Ketone-Based Metabolic Therapy: Is Increased NAD+ a Primary Mechanism?

    Directory of Open Access Journals (Sweden)

    Marwa Elamin

    2017-11-01

    Full Text Available The ketogenic diet’s (KD anticonvulsant effects have been well-documented for nearly a century, including in randomized controlled trials. Some patients become seizure-free and some remain so after diet cessation. Many recent studies have explored its expanded therapeutic potential in diverse neurological disorders, yet no mechanism(s of action have been established. The diet’s high fat, low carbohydrate composition reduces glucose utilization and promotes the production of ketone bodies. Ketone bodies are a more efficient energy source than glucose and improve mitochondrial function and biogenesis. Cellular energy production depends on the metabolic coenzyme nicotinamide adenine dinucleotide (NAD, a marker for mitochondrial and cellular health. Furthermore, NAD activates downstream signaling pathways (such as the sirtuin enzymes associated with major benefits such as longevity and reduced inflammation; thus, increasing NAD is a coveted therapeutic endpoint. Based on differential NAD+ utilization during glucose- vs. ketone body-based acetyl-CoA generation for entry into the tricarboxylic cycle, we propose that a KD will increase the NAD+/NADH ratio. When rats were fed ad libitum KD, significant increases in hippocampal NAD+/NADH ratio and blood ketone bodies were detected already at 2 days and remained elevated at 3 weeks, indicating an early and persistent metabolic shift. Based on diverse published literature and these initial data we suggest that increased NAD during ketolytic metabolism may be a primary mechanism behind the beneficial effects of this metabolic therapy in a variety of brain disorders and in promoting health and longevity.

  3. Metabolic Response to NAD Depletion across Cell Lines Is Highly Variable.

    Directory of Open Access Journals (Sweden)

    Yang Xiao

    Full Text Available Nicotinamide adenine dinucleotide (NAD is a cofactor involved in a wide range of cellular metabolic processes and is a key metabolite required for tumor growth. NAMPT, nicotinamide phosphoribosyltransferase, which converts nicotinamide (NAM to nicotinamide mononucleotide (NMN, the immediate precursor of NAD, is an attractive therapeutic target as inhibition of NAMPT reduces cellular NAD levels and inhibits tumor growth in vivo. However, there is limited understanding of the metabolic response to NAD depletion across cancer cell lines and whether all cell lines respond in a uniform manner. To explore this we selected two non-small cell lung carcinoma cell lines that are sensitive to the NAMPT inhibitor GNE-617 (A549, NCI-H1334, one that shows intermediate sensitivity (NCI-H441, and one that is insensitive (LC-KJ. Even though NAD was reduced in all cell lines there was surprising heterogeneity in their metabolic response. Both sensitive cell lines reduced glycolysis and levels of di- and tri-nucleotides and modestly increased oxidative phosphorylation, but they differed in their ability to combat oxidative stress. H1334 cells activated the stress kinase AMPK, whereas A549 cells were unable to activate AMPK as they contain a mutation in LKB1, which prevents activation of AMPK. However, A549 cells increased utilization of the Pentose Phosphate pathway (PPP and had lower reactive oxygen species (ROS levels than H1334 cells, indicating that A549 cells are better able to modulate an increase in oxidative stress. Inherent resistance of LC-KJ cells is associated with higher baseline levels of NADPH and a delayed reduction of NAD upon NAMPT inhibition. Our data reveals that cell lines show heterogeneous response to NAD depletion and that the underlying molecular and genetic framework in cells can influence the metabolic response to NAMPT inhibition.

  4. PGC1α-dependent NAD biosynthesis links oxidative metabolism to renal protection

    Science.gov (United States)

    Tran, Mei T.; Zsengeller, Zsuzsanna K.; Berg, Anders H.; Khankin, Eliyahu V.; Bhasin, Manoj K.; Kim, Wondong; Clish, Clary B.; Stillman, Isaac E.; Karumanchi, S. Ananth; Rhee, Eugene P.; Parikh, Samir M.

    2016-01-01

    The energetic burden of continuously concentrating solutes against gradients along the tubule may render the kidney especially vulnerable to ischemia. Indeed, acute kidney injury (AKI) affects 3% of all hospitalized patients.1,2 Here we show that the mitochondrial biogenesis regulator, PGC1α,3,4 is a pivotal determinant of renal recovery from injury by regulating NAD biosynthesis. Following renal ischemia, PGC1α−/− mice developed local deficiency of the NAD precursor niacinamide (Nam), marked fat accumulation, and failure to re-establish normal function. Remarkably, exogenous Nam improved local NAD levels, fat accumulation, and renal function in post-ischemic PGC1α−/− mice. Inducible tubular transgenic mice (iNephPGC1α) recapitulated the effects of Nam supplementation, including more local NAD and less fat accumulation with better renal function after ischemia. PGC1α coordinately upregulated the enzymes that synthesize NAD de novo from amino acids whereas PGC1α deficiency or AKI attenuated the de novo pathway. Nam enhanced NAD via the enzyme NAMPT and augmented production of the fat breakdown product beta-hydroxybutyrate (β-OHB), leading to increased prostaglandin PGE2, a secreted autocoid that maintains renal function.5 Nam treatment reversed established ischemic AKI and also prevented AKI in an unrelated toxic model. Inhibition of β-OHB signaling or prostaglandins similarly abolished PGC1α-dependent renoprotection. Given the importance of mitochondrial health in aging and the function of metabolically active organs, the results implicate Nam and NAD as key effectors for achieving PGC1α-dependent stress resistance. PMID:26982719

  5. NAD+ metabolite levels as a function of vitamins and calorie restriction: evidence for different mechanisms of longevity

    Directory of Open Access Journals (Sweden)

    Song Peng

    2010-02-01

    Full Text Available Abstract Background NAD+ is a coenzyme for hydride transfer enzymes and a substrate for sirtuins and other NAD+-dependent ADPribose transfer enzymes. In wild-type Saccharomyces cerevisiae, calorie restriction accomplished by glucose limitation extends replicative lifespan in a manner that depends on Sir2 and the NAD+ salvage enzymes, nicotinic acid phosphoribosyl transferase and nicotinamidase. Though alterations in the NAD+ to nicotinamide ratio and the NAD+ to NADH ratio are anticipated by models to account for the effects of calorie restriction, the nature of a putative change in NAD+ metabolism requires analytical definition and quantification of the key metabolites. Results Hydrophilic interaction chromatography followed by tandem electrospray mass spectrometry were used to identify the 12 compounds that constitute the core NAD+ metabolome and 6 related nucleosides and nucleotides. Whereas yeast extract and nicotinic acid increase net NAD+ synthesis in a manner that can account for extended lifespan, glucose restriction does not alter NAD+ or nicotinamide levels in ways that would increase Sir2 activity. Conclusions The results constrain the possible mechanisms by which calorie restriction may regulate Sir2 and suggest that provision of vitamins and calorie restriction extend lifespan by different mechanisms.

  6. Comparative Metabolomic Profiling Reveals That Dysregulated Glycolysis Stemming from Lack of Salvage NAD+ Biosynthesis Impairs Reproductive Development in Caenorhabditis elegans.

    Science.gov (United States)

    Wang, Wenqing; McReynolds, Melanie R; Goncalves, Jimmy F; Shu, Muya; Dhondt, Ineke; Braeckman, Bart P; Lange, Stephanie E; Kho, Kelvin; Detwiler, Ariana C; Pacella, Marisa J; Hanna-Rose, Wendy

    2015-10-23

    Temporal developmental progression is highly coordinated in Caenorhabditis elegans. However, loss of nicotinamidase PNC-1 activity slows reproductive development, uncoupling it from its typical progression relative to the soma. Using LC/MS we demonstrate that pnc-1 mutants do not salvage the nicotinamide released by NAD(+) consumers to resynthesize NAD(+), resulting in a reduction in global NAD(+) bioavailability. We manipulate NAD(+) levels to demonstrate that a minor deficit in NAD(+) availability is incompatible with a normal pace of gonad development. The NAD(+) deficit compromises NAD(+) consumer activity, but we surprisingly found no functional link between consumer activity and reproductive development. As a result we turned to a comparative metabolomics approach to identify the cause of the developmental phenotype. We reveal widespread metabolic perturbations, and using complementary pharmacological and genetic approaches, we demonstrate that a glycolytic block accounts for the slow pace of reproductive development. Interestingly, mitochondria are protected from both the deficiency in NAD(+) biosynthesis and the effects of reduced glycolytic output. We suggest that compensatory metabolic processes that maintain mitochondrial activity in the absence of efficient glycolysis are incompatible with the requirements for reproductive development, which requires high levels of cell division. In addition to demonstrating metabolic requirements for reproductive development, this work also has implications for understanding the mechanisms behind therapeutic interventions that target NAD(+) salvage biosynthesis for the purposes of inhibiting tumor growth. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

  7. Insight into molecular and functional properties of NMNAT3 reveals new hints of NAD homeostasis within human mitochondria.

    Directory of Open Access Journals (Sweden)

    Roberta Felici

    Full Text Available Among the enzymes involved in NAD homeostasis, nicotinamide mononucleotide adenylyltransferases (NMNAT1-3 are central to intracellular NAD formation. Although NMNAT3 is postulated to be a mitochondrial enzyme contributing to NAD-dependent organelle functioning, information on endogenous proteins is lacking. We report that in human cells a single gene nmnat3 localized on chromosome 3 codes for two mRNA splice variants NMNATv1 and FKSG76, whereas the previously reported NMNAT3v2 transcript is not present. However, NMNAT3v1 and FKSG76 proteins are not detectable, consistent with the finding that an upstream ORF in their mRNAs negatively regulates translation. NMNAT3v1 transfection demonstrates that the protein is cytosolic and inactive, whereas FKSG76 is mitochondrial but operates NAD cleavage rather than synthesis. In keeping with the lack of NMNAT3, we show that extracellular NAD, but not its metabolic precursors, sustains mitochondrial NAD pool in an ATP-independent manner. Data of the present study modify the scenario of the origin of mitochondrial NAD by showing that, in human cells, NMNAT3 is absent in mitochondria, and, akin to plants and yeast, cytosolic NAD maintains the mitochondrial NAD pool.

  8. Insight into molecular and functional properties of NMNAT3 reveals new hints of NAD homeostasis within human mitochondria.

    Science.gov (United States)

    Felici, Roberta; Lapucci, Andrea; Ramazzotti, Matteo; Chiarugi, Alberto

    2013-01-01

    Among the enzymes involved in NAD homeostasis, nicotinamide mononucleotide adenylyltransferases (NMNAT1-3) are central to intracellular NAD formation. Although NMNAT3 is postulated to be a mitochondrial enzyme contributing to NAD-dependent organelle functioning, information on endogenous proteins is lacking. We report that in human cells a single gene nmnat3 localized on chromosome 3 codes for two mRNA splice variants NMNATv1 and FKSG76, whereas the previously reported NMNAT3v2 transcript is not present. However, NMNAT3v1 and FKSG76 proteins are not detectable, consistent with the finding that an upstream ORF in their mRNAs negatively regulates translation. NMNAT3v1 transfection demonstrates that the protein is cytosolic and inactive, whereas FKSG76 is mitochondrial but operates NAD cleavage rather than synthesis. In keeping with the lack of NMNAT3, we show that extracellular NAD, but not its metabolic precursors, sustains mitochondrial NAD pool in an ATP-independent manner. Data of the present study modify the scenario of the origin of mitochondrial NAD by showing that, in human cells, NMNAT3 is absent in mitochondria, and, akin to plants and yeast, cytosolic NAD maintains the mitochondrial NAD pool.

  9. Formation of nicotinamide ribose diphosphate ribose, a new metabolite of the NAD pathway, by growing mycelium of Aspergillus niger

    International Nuclear Information System (INIS)

    Kuwahara, Masaaki

    1976-01-01

    A new step of NAD metabolism was shown in Aspergillus niger. Radioactive nicotinic acid and nicotinamide were incorporated into nicotinamide ribose diphosphate ribose (NAm-RDPR), which had been isolated from the culture filtrate. Its content in the culture medium increased with an increase of culture time, and this compound was proved to be a terminal metabolite in the NAD pathway. The experimental results also showed that the Preiss-Handler pathway and the NAD cycling system function in the NAD biosynthesis in A. niger. A part of the radioactive precursors was also incorporated into an unknown compound. (auth.)

  10. Neutron spectrometer using NE218 liquid scintillator

    International Nuclear Information System (INIS)

    Dance, J.B.; Francois, P.E.

    1976-01-01

    A neutron spectrometer has been constructed using NE218 liquid scintillator. Discrimination against electron-gamma events was obtained usng a charge-comparison pulse shape discrimination system. The resolution obtained was about 0.25 MeV F.W.H.M. at 2.0 MeV

  11. MiniBooNE overview and status

    Indian Academy of Sciences (India)

    P KAsPER (for the BooNE Collaboration). Fermilab, P.O. Box 500, Batavia, IL 60510-0500, USA. Abstract. Recent discoveries in the neutrino sector have opened a new frontier in high- energy physics and cosmology. Evidence from neutrino oscillation experiments from around the world indicate that neutrinos oscillate ...

  12. Mini Networked Screens (MiNeS)

    NARCIS (Netherlands)

    Prins, C.A.; Maris, M.; Breen, P.C.; Versteeg, N.; Terwisga, P.F. van; Ort, C.M.; Blok, J.J.

    2005-01-01

    The forward areas for an LPD in littoral waters can be full of surprises. A novel concept is presented for a networked screen consisting of elements of increasing capability to provide a progressive response to the threat. This MiNeS concept substantially improves the capability of the LPD as an

  13. The DAΦNE cryogenic system

    International Nuclear Information System (INIS)

    Modena, M.

    1997-12-01

    The DAΦNE Project utilises superconductivity technology for a total of six superconducting magnets: the two Experiment magnets (KLOE and FINUDA) and the four Compensator Solenoid magnets needed to compensate the magnetic effect of the Experiment magnets on the electron and positron beams. This effect, on beams of 510 MeV (nominal DAΦNE Energy), is expected to be relevant, especially with the aim of achieving a very high luminosity, which is the main target of the Project. The KLOE superconducting magnet has two possible working positions: the first in the DAΦNE Hall, when the Experiment will be in operation, and the second one in the KLOE Assembly Hall. This second position is the first to be utilised for the KLOE magnet Acceptance Test and magnetic field mapping, prior to the mounting of all the experimental apparatus inside the magnet. This note intends to present the DAΦNE Cryogenic System and how the authors have converged to the definition of a common Cryogenic System compatible with all the six superconducting magnets

  14. NAD+-Glycohydrolase Promotes Intracellular Survival of Group A Streptococcus.

    Directory of Open Access Journals (Sweden)

    Onkar Sharma

    2016-03-01

    Full Text Available A global increase in invasive infections due to group A Streptococcus (S. pyogenes or GAS has been observed since the 1980s, associated with emergence of a clonal group of strains of the M1T1 serotype. Among other virulence attributes, the M1T1 clone secretes NAD+-glycohydrolase (NADase. When GAS binds to epithelial cells in vitro, NADase is translocated into the cytosol in a process mediated by streptolysin O (SLO, and expression of these two toxins is associated with enhanced GAS intracellular survival. Because SLO is required for NADase translocation, it has been difficult to distinguish pathogenic effects of NADase from those of SLO. To resolve the effects of the two proteins, we made use of anthrax toxin as an alternative means to deliver NADase to host cells, independently of SLO. We developed a novel method for purification of enzymatically active NADase fused to an amino-terminal fragment of anthrax toxin lethal factor (LFn-NADase that exploits the avid, reversible binding of NADase to its endogenous inhibitor. LFn-NADase was translocated across a synthetic lipid bilayer in vitro in the presence of anthrax toxin protective antigen in a pH-dependent manner. Exposure of human oropharyngeal keratinocytes to LFn-NADase in the presence of protective antigen resulted in cytosolic delivery of NADase activity, inhibition of protein synthesis, and cell death, whereas a similar construct of an enzymatically inactive point mutant had no effect. Anthrax toxin-mediated delivery of NADase in an amount comparable to that observed during in vitro infection with live GAS rescued the defective intracellular survival of NADase-deficient GAS and increased the survival of SLO-deficient GAS. Confocal microscopy demonstrated that delivery of LFn-NADase prevented intracellular trafficking of NADase-deficient GAS to lysosomes. We conclude that NADase mediates cytotoxicity and promotes intracellular survival of GAS in host cells.

  15. NAD deamidation "a new reaction" by an enzyme from Aspergillus terreus DSM 826.

    Science.gov (United States)

    Elzainy, Tahany A; Ali, Thanaa H

    2005-02-01

    NAD deamidation is a non-previously recognized reaction. This reaction has been found to be catalyzed by extracts of Aspergillus terreus DSM 826. Conversion of NAD to the biosynthetic intermediate, deamido NAD, by these extracts, at the optimum pH and temperature did not exceed about 55 of the amount of the substrate added. Completion of the reaction was achieved when the extracts were pre-heated at 50 degrees C for 15 min in absence of the substrate. In a very similar manner, the extracts catalyzed hydrolytic cleavage of the amide linkages of different biomolecules such as nicotinamide, nicotinamide riboside, nicotinamide mononucleotide, L-glutamine, L-asparagine and acetamide. Polyacrylamide was also deamidated under the same conditions. In addition, complete dephosphorylation of the dinucleotide molecule was also effected by the same extracts. Separation of the NAD deamidating enzyme from the NAD dephosphorylating enzyme was achieved on using either DEAE - Sephadex A-25 or Sephadex G-200 column chromatography. The obtained phosphohydrolase-free-deamidase showed optimum activity at pH 8 of 0.1 M phosphate buffer and 50 degrees C. It exhibited broad substrate specificity and hyperbolic substrate saturation kinetics. It was isosterically inhibited by the product of its activity and this inhibition was prevented by heating the extracts at 50 degrees C for 15 min. Its activity was not affected in presence of sodium fluoride, partially inhibited in presence of magnesium chloride and was retained in the freezer for some months.

  16. NMN/NaMN adenylyltransferase (NMNAT) and NAD kinase (NADK) inhibitors: chemistry and potential therapeutic applications.

    Science.gov (United States)

    Petrelli, R; Felczak, K; Cappellacci, L

    2011-01-01

    Nicotinamide adenine dinucleotide (NAD(+)) has a crucial role in many cellular processes, both as a coenzyme for redox reactions and as a substrate to donate ADP-ribose units. Thus, enzymes involved in NAD(+) metabolism are attractive targets for drug discovery against a variety of human diseases. Herein we focus on two of them: NMN/NaMN adenylyltransferase (NMNAT) and NAD kinase (NADK). NMNAT is a key enzyme in all organisms catalyzing coupling of ATP and NMN or NaMN yielding NAD or NaAD, respectively. NADKs are ubiquitous enzymes involved in the last step of the biosynthesis of NADP. They phosphorylate NAD to produce NADP using ATP (or inorganic polyphosphates) in the presence of Mg(2+). No other pathway of NADP biosynthesis has been found in prokaryotic or eukaryotic cells. In this review we provide a comprehensive summary of NMNAT and NADK inhibitors highlighting their chemical modifications by different synthetic approaches, and structure-activity relationships depending on their potential therapeutic applications.

  17. Mobile phones electromagnetic radiation and NAD+-dependent isocitrate dehydrogenase as a mitochondrial marker in asthenozoospermia

    Directory of Open Access Journals (Sweden)

    Abeer M. Hagras

    2016-12-01

    Full Text Available NAD+-dependent Isocitrate Dehydrogenase (NAD+-IDH could be one of the cell phone radiation targets. Enzyme activity alteration may lead to decline in sperm motility during radio-frequency electromagnetic waves (RF-EMW exposure. The current case control study aimed to investigate the possible relationship between mitochondrial NAD+-IDH activity in human seminal plasma and sperm motility among asthenozoospermic cellular phone users. A total number of ninety idiopathic infertile males referred from the Department of Dermatology and Andrology, were enrolled in this study. NAD+-IDH activity was measured in human seminal plasma by spectrophotometer. Computer-aided sperm analysis (CASA following WHO criteria has been used for semen analyses. The results showed that IDH activity was increased in patients with prolonged cell phone daily use ≥4 h/day. Its level, correlated negatively with either the motility ratio percentages (r = −0.46, p < 0.001 or the progressive motility percentages (r = −0.50, p < 0.001 in the study groups. The current study suggests that NAD+-IDH in human seminal plasma could be one of seminal plasma biomarkers reflecting the mitochondrial function of spermatozoa. Alteration of its level could reflect the defective motility of sperms among some cases of cellular phone users.

  18. The plant mitochondrial mat-r gene/nad1 gene complex

    Energy Technology Data Exchange (ETDEWEB)

    Wolstenhome, D.R.

    1996-12-31

    We have completed sequencing segments of the maize mitochondrial (mt) DNA that contains all five of the exons (A-E) of the gene (nad1) for subunit I of the respiratory chain NADH dehydrogenase. Analysis of these sequences indicates that exons B and C are joined by a continuous group II intron, but the remaining exons are associated with partial group II introns and are encoded at widely separated locations in the maize mtDNA molecule. We have shown that mature transcripts of the maize nad1 gene contain 23 edited nucleotides, and that transcripts of maize and soybean mat-r genes contain 15 and 14 edits, respectively. The majority of edits in nad1 transcripts result in amino acid replacements that increase similarity between the maize NAD1 protein and NAD1 proteins of other plant species and of animal species. We found that the intron between exons b and c is not edited. From data obtained using PCR and sequencing we have shown that transcripts containing all possible exon combinations exist in maize mitochondria.

  19. Nivation forms and processes in unconsolidated sediments, NE Greenland

    DEFF Research Database (Denmark)

    Christiansen, Hanne Hvidtfeldt

    1998-01-01

    Nivation, Nivation Hollow, Nival Backwall Faliure, Active layer Interflow, Pronival alluvial fans, NE Greenland......Nivation, Nivation Hollow, Nival Backwall Faliure, Active layer Interflow, Pronival alluvial fans, NE Greenland...

  20. Isonicotinamide Enhances Sir2 Protein-mediated Silencing and Longevity in Yeast by Raising Intracellular NAD+ Concentration*

    Science.gov (United States)

    McClure, Julie M.; Wierman, Margaret B.; Maqani, Nazif; Smith, Jeffrey S.

    2012-01-01

    Sirtuins are an evolutionarily conserved family of NAD+-dependent protein deacetylases that function in the regulation of gene transcription, cellular metabolism, and aging. Their activity requires the maintenance of an adequate intracellular NAD+ concentration through the combined action of NAD+ biosynthesis and salvage pathways. Nicotinamide (NAM) is a key NAD+ precursor that is also a byproduct and feedback inhibitor of the deacetylation reaction. In Saccharomyces cerevisiae, the nicotinamidase Pnc1 converts NAM to nicotinic acid (NA), which is then used as a substrate by the NAD+ salvage pathway enzyme NA phosphoribosyltransferase (Npt1). Isonicotinamide (INAM) is an isostere of NAM that stimulates yeast Sir2 deacetylase activity in vitro by alleviating the NAM inhibition. In this study, we determined that INAM stimulates Sir2 through an additional mechanism in vivo, which involves elevation of the intracellular NAD+ concentration. INAM enhanced normal silencing at the rDNA locus but only partially suppressed the silencing defects of an npt1Δ mutant. Yeast cells grown in media lacking NA had a short replicative life span, which was extended by INAM in a SIR2-dependent manner and correlated with increased NAD+. The INAM-induced increase in NAD+ was strongly dependent on Pnc1 and Npt1, suggesting that INAM increases flux through the NAD+ salvage pathway. Part of this effect was mediated by the NR salvage pathways, which generate NAM as a product and require Pnc1 to produce NAD+. We also provide evidence suggesting that INAM influences the expression of multiple NAD+ biosynthesis and salvage pathways to promote homeostasis during stationary phase. PMID:22539348

  1. Isonicotinamide enhances Sir2 protein-mediated silencing and longevity in yeast by raising intracellular NAD+ concentration.

    Science.gov (United States)

    McClure, Julie M; Wierman, Margaret B; Maqani, Nazif; Smith, Jeffrey S

    2012-06-15

    Sirtuins are an evolutionarily conserved family of NAD(+)-dependent protein deacetylases that function in the regulation of gene transcription, cellular metabolism, and aging. Their activity requires the maintenance of an adequate intracellular NAD(+) concentration through the combined action of NAD(+) biosynthesis and salvage pathways. Nicotinamide (NAM) is a key NAD(+) precursor that is also a byproduct and feedback inhibitor of the deacetylation reaction. In Saccharomyces cerevisiae, the nicotinamidase Pnc1 converts NAM to nicotinic acid (NA), which is then used as a substrate by the NAD(+) salvage pathway enzyme NA phosphoribosyltransferase (Npt1). Isonicotinamide (INAM) is an isostere of NAM that stimulates yeast Sir2 deacetylase activity in vitro by alleviating the NAM inhibition. In this study, we determined that INAM stimulates Sir2 through an additional mechanism in vivo, which involves elevation of the intracellular NAD(+) concentration. INAM enhanced normal silencing at the rDNA locus but only partially suppressed the silencing defects of an npt1Δ mutant. Yeast cells grown in media lacking NA had a short replicative life span, which was extended by INAM in a SIR2-dependent manner and correlated with increased NAD(+). The INAM-induced increase in NAD(+) was strongly dependent on Pnc1 and Npt1, suggesting that INAM increases flux through the NAD(+) salvage pathway. Part of this effect was mediated by the NR salvage pathways, which generate NAM as a product and require Pnc1 to produce NAD(+). We also provide evidence suggesting that INAM influences the expression of multiple NAD(+) biosynthesis and salvage pathways to promote homeostasis during stationary phase.

  2. Lääne-Virumaa TOP 100 aastal 2000

    Index Scriptorium Estoniae

    2001-01-01

    Lääne-Virumaa edukamad ettevõtted; Lääne-Virumaa käibe TOP 100; Käibe kasvu TOP 20; Käibe languse TOP 10; Kasumi TOP 20; Kasumi kasvu TOP 20; Rentabluse TOP 20; ROA TOP 20; Kasumi languse TOP 10; Kahjumi TOP 10; Lääne-Virumaa käibelt suuremate ettevõtete finantsandmed. Lääne-Virumaa ettevõtete üldandmed

  3. Yrast and high spin states in 22Ne

    International Nuclear Information System (INIS)

    Szanto, E.M.; Toledo, A.S. de

    1982-08-01

    High spin states in 22 Ne have been investigated by the reactions 11 B( 13 C,d) 22 Ne and 13 C( 11 B,d) 22 Ne up to E* approximately=19 MeV. Yrast states were observed at 11.02 MeV (8 + ) and 15.46 MeV (10 + ) excitation energy. A backbending in 22 Ne is observed around spin 8 + . The location of high spin states I [pt

  4. NAD+ Is a Food Component That Promotes Exit from Dauer Diapause in Caenorhabditis elegans.

    Directory of Open Access Journals (Sweden)

    Mykola Mylenko

    Full Text Available The free-living soil nematode Caenorhabditis elegans adapts its development to the availability of food. When food is scarce and population density is high, worms enter a developmentally arrested non-feeding diapause stage specialized for long-term survival called the dauer larva. When food becomes available, they exit from the dauer stage, resume growth and reproduction. It has been postulated that compound(s present in food, referred to as the "food signal", promote exit from the dauer stage. In this study, we have identified NAD+ as a component of bacterial extract that promotes dauer exit. NAD+, when dissolved in alkaline medium, causes opening of the mouth and ingestion of food. We also show that to initiate exit from the dauer stage in response to NAD+ worms require production of serotonin. Thus, C. elegans can use redox cofactors produced by dietary organisms to sense food.

  5. Pharmacological NAD-Boosting Strategies Improve Mitochondrial Homeostasis in Human Complex I-Mutant Fibroblasts.

    Science.gov (United States)

    Felici, Roberta; Lapucci, Andrea; Cavone, Leonardo; Pratesi, Sara; Berlinguer-Palmini, Rolando; Chiarugi, Alberto

    2015-06-01

    Mitochondrial disorders are devastating genetic diseases for which efficacious therapies are still an unmet need. Recent studies report that increased availability of intracellular NAD obtained by inhibition of the NAD-consuming enzyme poly(ADP-ribose) polymerase (PARP)-1 or supplementation with the NAD-precursor nicotinamide riboside (NR) ameliorates energetic derangement and symptoms in mouse models of mitochondrial disorders. Whether these pharmacological approaches also improve bioenergetics of human cells harboring mitochondrial defects is unknown. It is also unclear whether the same signaling cascade is prompted by PARP-1 inhibitors and NR supplementation to improve mitochondrial homeostasis. Here, we show that human fibroblasts mutant for the NADH dehydrogenase (ubiquinone) Fe-S protein 1 (NDUFS1) subunit of respiratory complex I have similar ATP, NAD, and mitochondrial content compared with control cells, but show reduced mitochondrial membrane potential. Interestingly, mutant cells also show increased transcript levels of mitochondrial DNA but not nuclear DNA respiratory complex subunits, suggesting activation of a compensatory response. At variance with prior work in mice, however, NR supplementation, but not PARP-1 inhibition, increased intracellular NAD content in NDUFS1 mutant human fibroblasts. Conversely, PARP-1 inhibitors, but not NR supplementation, increased transcription of mitochondrial transcription factor A and mitochondrial DNA-encoded respiratory complexes constitutively induced in mutant cells. Still, both NR and PARP-1 inhibitors restored mitochondrial membrane potential and increased organelle content as well as oxidative activity of NDUFS1-deficient fibroblasts. Overall, data provide the first evidence that in human cells harboring a mitochondrial respiratory defect exposure to NR or PARP-1, inhibitors activate different signaling pathways that are not invariantly prompted by NAD increases, but equally able to improve energetic

  6. Ozone affects pollen viability and NAD(P)H oxidase release from Ambrosia artemisiifolia pollen

    Energy Technology Data Exchange (ETDEWEB)

    Pasqualini, Stefania, E-mail: spas@unipg.it [Department of Applied Biology, University of Perugia, Perugia (Italy); Tedeschini, Emma; Frenguelli, Giuseppe [Department of Applied Biology, University of Perugia, Perugia (Italy); Wopfner, Nicole; Ferreira, Fatima [Department of Molecular Biology, CD Laboratory for Allergy Diagnosis and Therapy, University of Salzburg, Salzburg (Austria); D' Amato, Gennaro [Division of Respiratory and Allergic Diseases, ' A. Cardarelli' High Speciality Hospital, Naples (Italy); Ederli, Luisa [Department of Applied Biology, University of Perugia, Perugia (Italy)

    2011-10-15

    Air pollution is frequently proposed as a cause of the increased incidence of allergy in industrialised countries. We investigated the impact of ozone (O{sub 3}) on reactive oxygen species (ROS) and allergen content of ragweed pollen (Ambrosia artemisiifolia). Pollen was exposed to acute O{sub 3} fumigation, with analysis of pollen viability, ROS and nitric oxide (NO) content, activity of nicotinamide adenine dinucleotide phosphate (NAD[P]H) oxidase, and expression of major allergens. There was decreased pollen viability after O{sub 3} fumigation, which indicates damage to the pollen membrane system, although the ROS and NO contents were not changed or were only slightly induced, respectively. Ozone exposure induced a significant enhancement of the ROS-generating enzyme NAD(P)H oxidase. The expression of the allergen Amb a 1 was not affected by O{sub 3}, determined from the mRNA levels of the major allergens. We conclude that O{sub 3} can increase ragweed pollen allergenicity through stimulation of ROS-generating NAD(P)H oxidase. - Highlights: > O{sub 3} reduces the viability of ragweed pollen. > ROS and allergens of ragweed pollen were not affected by O{sub 3} exposure. > O{sub 3} enhances the activity of the ROS-generating enzyme NAD(P)H oxidase. > O{sub 3} increases ragweed pollen allergenicity through NAD(P)H-oxidase stimulation. - This study focuses on the effects of the atmospheric pollutant ozone on ROS content and NAD(P)H oxidase activity of ragweed pollen grains.

  7. Dunnione ameliorates cisplatin-induced small intestinal damage by modulating NAD{sup +} metabolism

    Energy Technology Data Exchange (ETDEWEB)

    Pandit, Arpana; Kim, Hyung-Jin; Oh, Gi-Su; Shen, AiHua; Lee, Su-Bin; Khadka, Dipendra; Lee, SeungHoon [Center for Metabolic Function Regulation & Department of Microbiology, Wonkwang University, Iksan, Jeonbuk 570-749 (Korea, Republic of); Shim, Hyeok; Yang, Sei-Hoon; Cho, Eun-Young [Department of Internal Medicine, School of Medicine, Wonkwang University, Iksan, Jeonbuk 570-749 (Korea, Republic of); Kwon, Kang-Beom [Department of Oriental Medical Physiology, School of Oriental Medicine, Wonkwang University, Iksan, Jeonbuk 570-749 (Korea, Republic of); Kwak, Tae Hwan [PAEAN Biotechnology, 160 Techno-2 Street, Yuseong-gu, Daejeon 305-500 (Korea, Republic of); Choe, Seong-Kyu; Park, Raekil [Center for Metabolic Function Regulation & Department of Microbiology, Wonkwang University, Iksan, Jeonbuk 570-749 (Korea, Republic of); So, Hong-Seob, E-mail: jeanso@wku.ac.kr [Center for Metabolic Function Regulation & Department of Microbiology, Wonkwang University, Iksan, Jeonbuk 570-749 (Korea, Republic of)

    2015-11-27

    Although cisplatin is a widely used anticancer drug for the treatment of a variety of tumors, its use is critically limited because of adverse effects such as ototoxicity, nephrotoxicity, neuropathy, and gastrointestinal damage. Cisplatin treatment increases oxidative stress biomarkers in the small intestine, which may induce apoptosis of epithelial cells and thereby elicit damage to the small intestine. Nicotinamide adenine dinucleotide (NAD{sup +}) is a cofactor for various enzymes associated with cellular homeostasis. In the present study, we demonstrated that the hyper-activation of poly(ADP-ribose) polymerase-1 (PARP-1) is closely associated with the depletion of NAD{sup +} in the small intestine after cisplatin treatment, which results in downregulation of sirtuin1 (SIRT1) activity. Furthermore, a decrease in SIRT1 activity was found to play an important role in cisplatin-mediated small intestinal damage through nuclear factor (NF)-κB p65 activation, facilitated by its acetylation increase. However, use of dunnione as a strong substrate for the NADH:quinone oxidoreductase 1 (NQO1) enzyme led to an increase in intracellular NAD{sup +} levels and prevented the cisplatin-induced small intestinal damage correlating with the modulation of PARP-1, SIRT1, and NF-κB. These results suggest that direct modulation of cellular NAD{sup +} levels by pharmacological NQO1 substrates could be a promising therapeutic approach for protecting against cisplatin-induced small intestinal damage. - Highlights: • NAD{sup +} acts as a cofactor for numerous enzymes including Sirtuins and PARP. • Up-regulation of SIRT1 could attenuate the cisplatin-induced intestinal damage. • Modulation of the cellular NAD{sup +} could be a promising therapeutic approach.

  8. Regulation of the intersubunit ammonia tunnel in Mycobacterium tuberculosis glutamine-dependent NAD[superscript +] synthetase

    Energy Technology Data Exchange (ETDEWEB)

    Chuenchor, Watchalee; Doukov, Tzanko I.; Resto, Melissa; Chang, Andrew; Gerratana, Barbara (SSRL); (Maryland)

    2012-08-31

    Glutamine-dependent NAD{sup +} synthetase is an essential enzyme and a validated drug target in Mycobacterium tuberculosis (mtuNadE). It catalyses the ATP-dependent formation of NAD{sup +} from NaAD{sup +} (nicotinic acid-adenine dinucleotide) at the synthetase active site and glutamine hydrolysis at the glutaminase active site. An ammonia tunnel 40 {angstrom} (1 {angstrom} = 0.1 nm) long allows transfer of ammonia from one active site to the other. The enzyme displays stringent kinetic synergism; however, its regulatory mechanism is unclear. In the present paper, we report the structures of the inactive glutaminase C176A variant in an apo form and in three synthetase-ligand complexes with substrates (NaAD{sup +}/ATP), substrate analogue {l_brace}NaAD{sup +}/AMP-CPP (adenosine 5'-[{alpha},{beta}-methylene]triphosphate){r_brace} and intermediate analogues (NaAD{sup +}/AMP/PPi), as well as the structure of wild-type mtuNadE in a product complex (NAD{sup +}/AMP/PPi/glutamate). This series of structures provides snapshots of the ammonia tunnel during the catalytic cycle supported also by kinetics and mutagenesis studies. Three major constriction sites are observed in the tunnel: (i) at the entrance near the glutaminase active site; (ii) in the middle of the tunnel; and (iii) at the end near the synthetase active site. Variation in the number and radius of the tunnel constrictions is apparent in the crystal structures and is related to ligand binding at the synthetase domain. These results provide new insight into the regulation of ammonia transport in the intermolecular tunnel of mtuNadE.

  9. Identification of a magnesium-dependent NAD(P)(H)-binding domain in the nicotinoprotein methanol dehydrogenase from Bacillus methanolicus

    NARCIS (Netherlands)

    Hektor, HJ; Kloosterman, H; Dijkhuizen, L

    2002-01-01

    The Bacillus methanolicus methanol dehydrogenase (MDH) is a decameric nicotinoprotein alcohol dehydrogenase (family III) with one Zn2+ ion, one or two Mg2+ ions, and a tightly bound cofactor NAD(H) per subunit. The Mg2+ ions are essential for binding of cofactor NAD(H) in MDH. A B. methanolicus

  10. Defective mitophagy in XPA via PARP-1 hyperactivation and NAD(+)/SIRT1 reduction

    DEFF Research Database (Denmark)

    Fang, Evandro Fei; Scheibye-Knudsen, Morten; Brace, Lear E

    2014-01-01

    with excessive cleavage of PINK1 and increased mitochondrial membrane potential. The mitochondrial abnormalities appear to be caused by decreased activation of the NAD(+)-SIRT1-PGC-1α axis triggered by hyperactivation of the DNA damage sensor PARP-1. This phenotype is rescued by PARP-1 inhibition...... or by supplementation with NAD(+) precursors that also rescue the lifespan defect in xpa-1 nematodes. Importantly, this pathogenesis appears common to ataxia-telangiectasia and Cockayne syndrome, two other DNA repair disorders with neurodegeneration, but absent in XPC, a DNA repair disorder without neurodegeneration...

  11. Status of KM3NeT

    Directory of Open Access Journals (Sweden)

    Riccobene G.

    2016-01-01

    Full Text Available The recent observation of cosmic neutrinos by IceCube has pushed the quest towards the identification of cosmic sources of high-energy particles. The KM3NeT Collaboration is now ready to launch the massive construction of detection units to be installed in deep sea to build a km-cubic size neutrino telescope. The main elements of the detector, the status of the project and the expected perfomances are briefly reported.

  12. NAD+-Dependent Activation of Sirt1 Corrects the Phenotype in a Mouse Model of Mitochondrial Disease

    Science.gov (United States)

    Cerutti, Raffaele; Pirinen, Eija; Lamperti, Costanza; Marchet, Silvia; Sauve, Anthony A.; Li, Wei; Leoni, Valerio; Schon, Eric A.; Dantzer, Françoise; Auwerx, Johan; Viscomi, Carlo; Zeviani, Massimo

    2014-01-01

    Summary Mitochondrial disorders are highly heterogeneous conditions characterized by defects of the mitochondrial respiratory chain. Pharmacological activation of mitochondrial biogenesis has been proposed as an effective means to correct the biochemical defects and ameliorate the clinical phenotype in these severely disabling, often fatal, disorders. Pathways related to mitochondrial biogenesis are targets of Sirtuin1, a NAD+-dependent protein deacetylase. As NAD+ boosts the activity of Sirtuin1 and other sirtuins, intracellular levels of NAD+ play a key role in the homeostatic control of mitochondrial function by the metabolic status of the cell. We show here that supplementation with nicotinamide riboside, a natural NAD+ precursor, or reduction of NAD+ consumption by inhibiting the poly(ADP-ribose) polymerases, leads to marked improvement of the respiratory chain defect and exercise intolerance of the Sco2 knockout/knockin mouse, a mitochondrial disease model characterized by impaired cytochrome c oxidase biogenesis. This strategy is potentially translatable into therapy of mitochondrial disorders in humans. PMID:24814483

  13. High-resolution structures of Thermus thermophilus enoyl-acyl carrier protein reductase in the apo form, in complex with NAD+ and in complex with NAD+ and triclosan

    International Nuclear Information System (INIS)

    Otero, José M.; Noël, Ann-Josée; Guardado-Calvo, Pablo; Llamas-Saiz, Antonio L.; Wende, Wolfgang; Schierling, Benno; Pingoud, Alfred; Raaij, Mark J. van

    2012-01-01

    T. thermophilus enoyl-acyl carrier protein reductase was crystallized in the apo form, with NAD + bound and with NAD + and the inhibitor triclosan bound. The structures were solved by molecular replacement and refined at 1.50, 1.86 and 1.90 Å resolution, respectively. The structures are described, analysed and compared with those of enoyl-acyl carrier protein reductases from other species. Enoyl-acyl carrier protein reductase (ENR; the product of the fabI gene) is an important enzyme that is involved in the type II fatty-acid-synthesis pathway of bacteria, plants, apicomplexan protozoa and mitochondria. Harmful pathogens such as Mycobacterium tuberculosis and Plasmodium falciparum use the type II fatty-acid-synthesis system, but not mammals or fungi, which contain a type I fatty-acid-synthesis pathway consisting of one or two multifunctional enzymes. For this reason, specific inhibitors of ENR are attractive antibiotic candidates. Triclosan, a broad-range antibacterial agent, binds to ENR, inhibiting fatty-acid synthesis. As humans do not have an ENR enzyme, they are not affected. Here, high-resolution structures of Thermus thermophilus (Tth) ENR in the apo form, bound to NAD + and bound to NAD + plus triclosan are reported. Differences from and similarities to other known ENR structures are reported; in general, the structures are very similar. The cofactor-binding site is also very similar to those of other ENRs and, as reported for other species, triclosan leads to greater ordering of the loop that covers the cofactor-binding site, which, together with the presence of triclosan itself, presumably provides tight binding of the dinucleotide, preventing cycling of the cofactor. Differences between the structures of Tth ENR and other ENRs are the presence of an additional β-sheet at the N-terminus and a larger number of salt bridges and side-chain hydrogen bonds. These features may be related to the high thermal stability of Tth ENR

  14. Performance Analysis of Different NeQuick Ionospheric Model Parameters

    Directory of Open Access Journals (Sweden)

    WANG Ningbo

    2017-04-01

    Full Text Available Galileo adopts NeQuick model for single-frequency ionospheric delay corrections. For the standard operation of Galileo, NeQuick model is driven by the effective ionization level parameter Az instead of the solar activity level index, and the three broadcast ionospheric coefficients are determined by a second-polynomial through fitting the Az values estimated from globally distributed Galileo Sensor Stations (GSS. In this study, the processing strategies for the estimation of NeQuick ionospheric coefficients are discussed and the characteristics of the NeQuick coefficients are also analyzed. The accuracy of Global Position System (GPS broadcast Klobuchar, original NeQuick2 and fitted NeQuickC as well as Galileo broadcast NeQuickG models is evaluated over the continental and oceanic regions, respectively, in comparison with the ionospheric total electron content (TEC provided by global ionospheric maps (GIM, GPS test stations and JASON-2 altimeter. The results show that NeQuickG can mitigate ionospheric delay by 54.2%~65.8% on a global scale, and NeQuickC can correct for 71.1%~74.2% of the ionospheric delay. NeQuick2 performs at the same level with NeQuickG, which is a bit better than that of GPS broadcast Klobuchar model.

  15. Estimation of Avalanche Hazard in the Settlement of Magurka using Elba+ Model / Posúdenie Ohrozenosti Osady Magurky Lavínami S Použitím Modelu Elba+

    Directory of Open Access Journals (Sweden)

    Bartík Martin

    2013-06-01

    Full Text Available V našej študií sme sa zamerali na moderné softvérové aplikácie umožňujúce simuláciu lavíny. Použili sme model ELBA+, pomocou ktorého sme snažili zhodnotiť ohrozenosť horského prostredia v okolí starej banskej osady Magurka (1 036 m n. m., ktorá leží v závere Ľupčianskej doline pod hlavným hrebeňom Nízkych Tatier. Do histórie sa zapísala hlavne lavína v doline Ďurková zo 14. marca 1970, ktorá dodnes patrí medzi najväčšie lavíny zaznamenané na Slovensku. S použitím archívnych údajov Strediska lavínovej prevencie v Jasnej sme sa pokúsili o jej čo najvernejšiu rekonštrukciu. Následne sme sa pokúsili o simuláciu lavíny v doline Viedenka pri použití rovnakej výšky snehu v odtrhovom pásme ako sa predpokladá pri lavíne v roku 1970. Keďže časť osady Magurka je situovaných pri ústí tejto doliny, ktorá má priamejší aj kratší priebeh ako dolina Ďurková, skúmali sme možnosť jej zásahu lavínou.

  16. Sterile Neutrino Searches in MiniBooNE and MicroBooNE

    Energy Technology Data Exchange (ETDEWEB)

    Ignarra, Christina M. [Massachusetts Inst. of Technology (MIT), Cambridge, MA (United States)

    2014-09-01

    Tension among recent short baseline neutrino experiments has pointed toward the possible need for the addition of one or more sterile (non-interacting) neutrino states into the existing neutrino oscillation framework. This thesis first presents the motivation for sterile neutrino models by describing the short-baseline anomalies that can be addressed with them. This is followed by a discussion of the phenomenology of these models. The MiniBooNE experiment and results are then described in detail, particularly the most recent antineutrino analysis. This will be followed by a discussion of global fits to world data, including the anomalous data sets. Lastly, future experiments will be addressed, especially focusing on the MicroBooNE experiment and light collection studies. In particular, understanding the degradation source of TPB, designing the TPB-coated plates for MicroBooNE and developing lightguide collection systems will be discussed. We find an excess of events in the MiniBooNE antineutrino mode results consistent with the LSND anomaly, but one that has a different energy dependence than the low-energy excess reported in neutrino mode. This disagreement creates tension within global fits which include up to three sterile neutrinos. The low-energy excess will be addressed by the MicroBooNE experiment, which is expected to start taking data in early 2015. Tension among existing experiments calls for additional, more decisive future experiments.

  17. Sterile Neutrino Searches in MiniBooNE and MicroBooNE

    Energy Technology Data Exchange (ETDEWEB)

    Ignarra, Christina M. [Massachusetts Inst. of Technology (MIT), Cambridge, MA (United States)

    2014-09-01

    Tension among recent short baseline neutrino experiments has pointed toward the possible need for the addition of one or more sterile (non-interacting) neutrino states into the existing neutrino oscillation framework. This thesis fi rst presents the motivation for sterile neutrino models by describing the short-baseline anomalies that can be addressed with them. This is followed by a discussion of the phenomenology of these models. The MiniBooNE experiment and results are then described in detail, particularly the most recent antineutrino analysis. This will be followed by a discussion of global fits to world data, including the anomalous data sets. Lastly, future experiments will be addressed, especially focusing on the MicroBooNE experiment and light collection studies. In particular, understanding the degradation source of TPB, designing the TPB-coated plates for MicroBooNE and developing lightguide collection systems will be discussed. We find an excess of events in the MiniBooNE antineutrino mode results consistent with the LSND anomaly, but one that has a di fferent energy dependence than the low-energy excess reported in neutrino mode. This disagreement creates tension within global fi ts which include up to three sterile neutrinos. The low-energy excess will be addressed by the MicroBooNE experiment, which is expected to start taking data in early 2015. Tension among existing experiments calls for additional, more decisive future experiments.

  18. Homogeneous nucleation rates of nitric acid dihydrate (NAD at simulated stratospheric conditions – Part II: Modelling

    Directory of Open Access Journals (Sweden)

    O. Möhler

    2006-01-01

    Full Text Available Activation energies ΔGact for the nucleation of nitric acid dihydrate (NAD in supercooled binary HNO3/H2O solution droplets were calculated from volume-based nucleation rate measurements using the AIDA (Aerosol, Interactions, and Dynamics in the Atmosphere aerosol chamber of Forschungszentrum Karlsruhe. The experimental conditions covered temperatures T between 192 and 197 K, NAD saturation ratios SNAD between 7 and 10, and nitric acid molar fractions of the nucleating sub-micron sized droplets between 0.26 and 0.28. Based on classical nucleation theory, a new parameterisation for ΔGact=A×(T ln SNAD−2+B is fitted to the experimental data with A=2.5×106 kcal K2 mol−1 and B=11.2−0.1(T−192 kcal mol−1. A and B were chosen to also achieve good agreement with literature data of ΔGact. The parameter A implies, for the temperature and composition range of our analysis, a mean interface tension σsl=51 cal mol−1 cm−2 between the growing NAD germ and the supercooled solution. A slight temperature dependence of the diffusion activation energy is represented by the parameter B. Investigations with a detailed microphysical process model showed that literature formulations of volume-based (Salcedo et al., 2001 and surface-based (Tabazadeh et al., 2002 nucleation rates significantly overestimate NAD formation rates when applied to the conditions of our experiments.

  19. Dinucleoside polyphosphate NAD analogs as potential NMN adenylyltransferase inhibitors. Synthesis and biological evaluation.

    Science.gov (United States)

    Franchetti, P; Cappellacci, L; Pasqualini, M; Grifantini, M; Lorenzi, T; Raffaelli, N; Magni, G

    2003-01-01

    Two dinucleoside polyphosphate NAD analogs, P1-(adenosine-5')-P3-(nicotinamide riboside-5')triphosphate (Np3A, 1) and P1-(adenosine-5')-P4-(nicotinamide riboside-5')tetraphosphate (Np4A, 2), were synthesized and tested as inhibitors of both microbial and human recombinant NMN adenylyltransferase. Compounds 1 and 2 proved to be selective inhibitors of microbial enzymes.

  20. Synthesis and biological evaluation of NAD analogs as human pyridine nucleotide adenylyltransferase inhibitors.

    Science.gov (United States)

    Franchetti, Palmarisa; Petrelli, R; Cappellacci, L; Pasqualini, M; Vita, P; Sorci, L; Mazzola, F; Raffaelli, N; Magni, Giulio

    2005-01-01

    NAD analogs modified at the ribose adenylyl moiety, named N-2'-MeAD and Na-2'-MeAD, were synthesized as ligands of pyridine nucleotide (NMN/NaMN) adenylyltransferase (NMNAT). Both dinucleotides resulted selective inhibitors against human NMNAT-3 isoenzyme.

  1. NAD+-dependent SIRT1 deacetylase participates in epigenetic reprogramming during endotoxin tolerance.

    Science.gov (United States)

    Liu, Tie Fu; Yoza, Barbara K; El Gazzar, Mohamed; Vachharajani, Vidula T; McCall, Charles E

    2011-03-18

    Gene-selective epigenetic reprogramming and shifts in cellular bioenergetics develop when Toll-like receptors (TLR) recognize and respond to systemic life-threatening infections. Using a human monocyte cell model of endotoxin tolerance and human leukocytes from acute systemic inflammation with sepsis, we report that energy sensor sirtuin 1 (SIRT1) coordinates the epigenetic and bioenergy shifts. After TLR4 signaling, SIRT1 rapidly accumulated at the promoters of TNF-α and IL-1β, but not IκBα; SIRT1 promoter binding was dependent on its co-factor, NAD(+). During this initial process, SIRT1 deacetylated RelA/p65 lysine 310 and nucleosomal histone H4 lysine 16 to promote termination of NFκB-dependent transcription. SIRT1 then remained promoter bound and recruited de novo induced RelB, which directed assembly of the mature transcription repressor complex that generates endotoxin tolerance. SIRT1 also promoted de novo expression of RelB. During sustained endotoxin tolerance, nicotinamide phosphoribosyltransferase (Nampt), the rate-limiting enzyme for endogenous production of NAD(+), and SIRT1 expression increased. The elevation of SIRT1 required protein stabilization and enhanced translation. To support the coordination of bioenergetics in human sepsis, we observed elevated NAD(+) levels concomitant with SIRT1 and RelB accumulation at the TNF-α promoter of endotoxin tolerant sepsis blood leukocytes. We conclude that TLR4 stimulation and human sepsis activate pathways that couple NAD(+) and its sensor SIRT1 with epigenetic reprogramming.

  2. Recycling of NAD(P) by multienzyme systems immobilized by microencapsulation in artificial cells.

    Science.gov (United States)

    Chang, T M

    1987-01-01

    Multistep enzyme systems can be immobilized in solution within semipermeable microcapsules. With the ability to recycle cofactors, a number of potentially useful systems have been made possible. Furthermore NAD+ can be retained inside the microcapsules by two approaches. (1) NAD+ can be linked to macromolecules such as dextran or polyethyleneimine. However, in this form, there are significant increases in steric hindrance and diffusion restrictions. (2) "Artificial cells" consisting of lipid-polyamide membrane microcapsules containing multienzyme systems, cofactors, and substrates can retain NAD+ in the free form. Analogous to the intracellular environments of red blood cells, free NAD+ in solution inside the microcapsules is effectively recycled by the multistep enzyme systems which are also in solution. Enzymes in the microcapsules are in high concentrations and in close proximity to one another. Any number and any concentration of different enzyme systems can be microencapsulated all within one artificial cell, within the limit of solubility of the total amount of enzymes. Products of sequential reactions inside the microcapsules are at much higher concentrations than outside. All these factors result in an optimal intracellular environment for multistep enzyme reactions. External substrates in the form of lipophilic or small hydrophilic molecules can equilibrate across the membrane to participate as initial substrates in the multistep reactions in the microcapsules. A number of potential applications are possible using this approach. The lipid-polyamide membrane artificial cell can also be used in basic research as a biochemical cell model for the simpler types of biological cells such as erythrocytes.

  3. Pancreatic Beta-Cell Purification by Altering FAD and NAD(PH Metabolism

    Directory of Open Access Journals (Sweden)

    P. de Vos

    2008-07-01

    Full Text Available Isolation of primary beta cells from other cells within in the pancreatic islets is of importance for many fields of islet research. However, up to now, no satisfactory method has been developed that gained high numbers of viable beta cells, without considerable alpha-cell contamination. In this study, we investigated whether rat beta cells can be isolated from nonbeta endocrine cells by manipulating the flavin adenine dinucleotide (FAD and nicotinamide-adenine dinucleotide phosphate (NAD(PH autofluorescence. Beta cells were isolated from dispersed islets by flow cytometry, based on their high FAD and NAD(PH fluorescence. To improve beta cell yield and purity, the cellular FAD and NAD(PH contents were altered by preincubation in culture media containing varying amounts of D-glucose and amino acids. Manipulation of the cellular FAD and NAD(PH fluorescence improves beta cell yield and purity after sorting. This method is also a fast and reliable method to measure beta cell functional viability. A conceivable application is assessing beta cell viability before transplantation.

  4. Clinical heterogeneity of mitochondrial NAD kinase deficiency caused by a NADK2 start loss variant

    NARCIS (Netherlands)

    Pomerantz, Daniel J.; Ferdinandusse, Sacha; Cogan, Joy; Cooper, David N.; Reimschisel, Tyler; Robertson, Amy; Bican, Anna; McGregor, Tracy; Gauthier, Jackie; Millington, David S.; Andrae, Jaime L. W.; Tschannen, Michael R.; Helbling, Daniel C.; Demos, Wendy M.; Denis, Simone; Wanders, Ronald J. A.; Newman, John N.; Hamid, Rizwan; Phillips, John A.

    2018-01-01

    Mitochondrial NAD kinase deficiency (NADK2D, OMIM #615787) is a rare autosomal recessive disorder of NADPH biosynthesis that can cause hyperlysinemia and dienoyl-CoA reductase deficiency (DECRD, OMIM #616034). NADK2 deficiency has been reported in only three unrelated patients. Two had severe,

  5. NAD(P)H regeneration is the key for heterolactic fermentation of hexoses in Oenococcus oeni.

    Science.gov (United States)

    Maicas, Sergi; Ferrer, Sergi; Pardo, Isabel

    2002-01-01

    Oenococcus oeni (formerly Leuconostoc oenos) can perform malolactic fermentation, converting L-malate to L-lactate and carbon dioxide, in wines. The energy and redox potential required to support the growth of the micro-organism are supplied mainly by the consumption of carbohydrates via the heterolactic pathway. In the first steps of hexose metabolism two molecules of NAD(P)(+) are consumed, which must be regenerated in later reactions. The aim of this work was to test if aerobic growth of O. oeni promotes higher cell yields than anaerobic conditions, as has been shown for other lactic acid bacteria. O. oeni M42 was found to grow poorly under aerobic conditions with glucose as the only carbohydrate in the medium. It was demonstrated that O(2) inactivates the enzymes of the ethanol-forming pathway, one of the two pathways which reoxidizes NAD(P)(+) cofactors in the heterolactic catabolism of glucose. These results suggest that the regeneration of cofactors is the limiting factor for the aerobic consumption of glucose. When external electron acceptors, such as fructose or pyruvate, were added to glucose-containing culture medium the growth of O. oeni was stimulated slightly; fructose was converted to mannitol, oxidizing two molecules of NAD(P)H, and pyruvate was transformed to lactate, enabling the regeneration of NAD(+). The addition of cysteine seemed to suppress the inactivation of the ethanol-forming pathway enzymes by O(2), enabling glucose consumption in aerobic conditions to reach similar rates to those found in anaerobic conditions.

  6. NAD(+) Replenishment Improves Lifespan and Healthspan in Ataxia Telangiectasia Models via Mitophagy and DNA Repair

    DEFF Research Database (Denmark)

    Fang, Evandro Fei; Kassahun, Henok; Croteau, Deborah L

    2016-01-01

    function, delay memory loss, and extend lifespan in both animal models. Mechanistically, treatments that increase intracellular NAD(+) also stimulate neuronal DNA repair and improve mitochondrial quality via mitophagy. This work links two major theories on aging, DNA damage accumulation, and mitochondrial...

  7. Prezentace zříceniny hradu Kamýku nad Vltavou

    Czech Academy of Sciences Publication Activity Database

    Durdík, Tomáš; Girsa, V.; Hanzl, M.

    2010-01-01

    Roč. 70, č. 3 (2010), 175-178,231,233-234 ISSN 1210-5538 R&D Projects: GA MK DB06P01OPP004 Institutional research plan: CEZ:AV0Z80020508 Keywords : castle * castellology * Bohemia * Kamýk nad Vltavou * medieval archaeology * architecture * monument care * presentation Subject RIV: AC - Archeology, Anthropology, Ethnology

  8. Live cell imaging of cytosolic NADH/NAD+ ratio in hepatocytes and liver slices.

    Science.gov (United States)

    Masia, Ricard; McCarty, William J; Lahmann, Carolina; Luther, Jay; Chung, Raymond T; Yarmush, Martin L; Yellen, Gary

    2018-01-01

    Fatty liver disease (FLD), the most common chronic liver disease in the United States, may be caused by alcohol or the metabolic syndrome. Alcohol is oxidized in the cytosol of hepatocytes by alcohol dehydrogenase (ADH), which generates NADH and increases cytosolic NADH/NAD + ratio. The increased ratio may be important for development of FLD, but our ability to examine this question is hindered by methodological limitations. To address this, we used the genetically encoded fluorescent sensor Peredox to obtain dynamic, real-time measurements of cytosolic NADH/NAD + ratio in living hepatocytes. Peredox was expressed in dissociated rat hepatocytes and HepG2 cells by transfection, and in mouse liver slices by tail-vein injection of adeno-associated virus (AAV)-encoded sensor. Under control conditions, hepatocytes and liver slices exhibit a relatively low (oxidized) cytosolic NADH/NAD + ratio as reported by Peredox. The ratio responds rapidly and reversibly to substrates of lactate dehydrogenase (LDH) and sorbitol dehydrogenase (SDH). Ethanol causes a robust dose-dependent increase in cytosolic NADH/NAD + ratio, and this increase is mitigated by the presence of NAD + -generating substrates of LDH or SDH. In contrast to hepatocytes and slices, HepG2 cells exhibit a relatively high (reduced) ratio and show minimal responses to substrates of ADH and SDH. In slices, we show that comparable results are obtained with epifluorescence imaging and two-photon fluorescence lifetime imaging (2p-FLIM). Live cell imaging with Peredox is a promising new approach to investigate cytosolic NADH/NAD + ratio in hepatocytes. Imaging in liver slices is particularly attractive because it allows preservation of liver microanatomy and metabolic zonation of hepatocytes. NEW & NOTEWORTHY We describe and validate a new approach for measuring free cytosolic NADH/NAD + ratio in hepatocytes and liver slices: live cell imaging with the fluorescent biosensor Peredox. This approach yields dynamic, real

  9. Role of residues in the adenosine binding site of NAD of the Ascaris suum malic enzyme.

    Science.gov (United States)

    Aktas, Deniz F; Cook, Paul F

    2008-12-01

    Ascaris suum mitochondrial malic enzyme catalyzes the divalent metal ion dependent conversion of l-malate to pyruvate and CO(2), with concomitant reduction of NAD(P) to NAD(P)H. In this study, some of the residues that form the adenosine binding site of NAD were mutated to determine their role in binding of the cofactor and/or catalysis. D361, which is completely conserved among species, is located in the dinucleotide-binding Rossmann fold and makes a salt bridge with R370, which is also highly conserved. D361 was mutated to E, A and N. R370 was mutated to K and A. D361E and A mutant enzymes were inactive, likely a result of the increase in the volume in the case of the D361E mutant enzyme that caused clashes with the surrounding residues, and loss of the ionic interaction between D361 and R370, for D361A. Although the K(m) for the substrates and isotope effect values did not show significant changes for the D361N mutant enzyme, V/E(t) decreased by 1400-fold. Data suggested the nonproductive binding of the cofactor, giving a low fraction of active enzyme. The R370K mutant enzyme did not show any significant changes in the kinetic parameters, while the R370A mutant enzyme gave a slight change in V/E(t), contrary to expectations. Overall, results suggest that the salt bridge between D361 and R370 is important for maintaining the productive conformation of the NAD binding site. Mutation of residues involved leads to nonproductive binding of NAD. The interaction stabilizes one of the Rossmann fold loops that NAD binds. Mutation of H377 to lysine, which is conserved in NADP-specific malic enzymes and proposed to be a cofactor specificity determinant, did not cause a shift in cofactor specificity of the Ascaris malic enzyme from NAD to NADP. However, it is confirmed that this residue is an important second layer residue that affects the packing of the first layer residues that directly interact with the cofactor.

  10. Nicotinamidase modulation of NAD+ biosynthesis and nicotinamide levels separately affect reproductive development and cell survival in C. elegans.

    Science.gov (United States)

    Vrablik, Tracy L; Huang, Li; Lange, Stephanie E; Hanna-Rose, Wendy

    2009-11-01

    Nicotinamide adenine dinucleotide (NAD(+)) is a central molecule in cellular metabolism and an obligate co-substrate for NAD(+)-consuming enzymes, which regulate key biological processes such as longevity and stress responses. Although NAD(+) biosynthesis has been intensely studied, little analysis has been done in developmental models. We have uncovered novel developmental roles for a nicotinamidase (PNC), the first enzyme in the NAD(+) salvage pathway of invertebrates. Mutations in the Caenorhabditis elegans nicotinamidase PNC-1 cause developmental and functional defects in the reproductive system; the development of the gonad is delayed, four uterine cells die by necrosis and the mutant animals are egg-laying defective. The temporal delay in gonad development results from depletion of the salvage pathway product NAD(+), whereas the uv1 cell necrosis and egg-laying defects result from accumulation of the substrate nicotinamide. Thus, regulation of both substrate and product level is key to the biological activity of PNC-1. We also find that diet probably affects the levels of these metabolites, as it affects phenotypes. Finally, we identified a secreted isoform of PNC-1 and confirmed its extracellular localization and functional activity in vivo. We demonstrate that nicotinamide phosphoribosyltransferase (Nampt), the equivalent enzyme in nicotinamide recycling to NAD(+) in vertebrates, can functionally substitute for PNC-1. As Nampt is also secreted, we postulate an evolutionarily conserved extracellular role for NAD(+) biosynthetic enzymes during development and physiology.

  11. Novel assay for simultaneous measurement of pyridine mononucleotides synthesizing activities allows dissection of the NAD(+) biosynthetic machinery in mammalian cells.

    Science.gov (United States)

    Zamporlini, Federica; Ruggieri, Silverio; Mazzola, Francesca; Amici, Adolfo; Orsomando, Giuseppe; Raffaelli, Nadia

    2014-11-01

    The redox coenzyme NAD(+) is also a rate-limiting co-substrate for several enzymes that consume the molecule, thus rendering its continuous re-synthesis indispensable. NAD(+) biosynthesis has emerged as a therapeutic target due to the relevance of NAD(+) -consuming reactions in complex intracellular signaling networks whose alteration leads to many neurologic and metabolic disorders. Distinct metabolic routes, starting from various precursors, are known to support NAD(+) biosynthesis with tissue/cell-specific efficiencies, probably reflecting differential expression of the corresponding rate-limiting enzymes, i.e. nicotinamide phosphoribosyltransferase, quinolinate phosphoribosyltransferase, nicotinate phosphoribosyltransferase and nicotinamide riboside kinase. Understanding the contribution of these enzymes to NAD(+) levels depending on the tissue/cell type and metabolic status is necessary for the rational design of therapeutic strategies aimed at modulating NAD(+) availability. Here we report a simple, fast and sensitive coupled fluorometric assay that enables simultaneous determination of the four activities in whole-cell extracts and biological fluids. Its application to extracts from various mouse tissues, human cell lines and plasma yielded for the first time an overall picture of the tissue/cell-specific distribution of the activities of the various enzymes. The screening enabled us to gather novel findings, including (a) the presence of quinolinate phosphoribosyltransferase and nicotinamide riboside kinase in all examined tissues/cell lines, indicating that quinolinate and nicotinamide riboside are relevant NAD(+) precursors, and (b) the unexpected occurrence of nicotinate phosphoribosyltransferase in human plasma. © 2014 FEBS.

  12. Tributyltin induces G2/M cell cycle arrest via NAD(+)-dependent isocitrate dehydrogenase in human embryonic carcinoma cells.

    Science.gov (United States)

    Asanagi, Miki; Yamada, Shigeru; Hirata, Naoya; Itagaki, Hiroshi; Kotake, Yaichiro; Sekino, Yuko; Kanda, Yasunari

    2016-04-01

    Organotin compounds, such as tributyltin (TBT), are well-known endocrine-disrupting chemicals (EDCs). We have recently reported that TBT induces growth arrest in the human embryonic carcinoma cell line NT2/D1 at nanomolar levels by inhibiting NAD(+)-dependent isocitrate dehydrogenase (NAD-IDH), which catalyzes the irreversible conversion of isocitrate to α-ketoglutarate. However, the molecular mechanisms by which NAD-IDH mediates TBT toxicity remain unclear. In the present study, we examined whether TBT at nanomolar levels affects cell cycle progression in NT2/D1 cells. Propidium iodide staining revealed that TBT reduced the ratio of cells in the G1 phase and increased the ratio of cells in the G2/M phase. TBT also reduced cell division cycle 25C (cdc25C) and cyclin B1, which are key regulators of G2/M progression. Furthermore, apigenin, an inhibitor of NAD-IDH, mimicked the effects of TBT. The G2/M arrest induced by TBT was abolished by NAD-IDHα knockdown. Treatment with a cell-permeable α-ketoglutarate analogue recovered the effect of TBT, suggesting the involvement of NAD-IDH. Taken together, our data suggest that TBT at nanomolar levels induced G2/M cell cycle arrest via NAD-IDH in NT2/D1 cells. Thus, cell cycle analysis in embryonic cells could be used to assess cytotoxicity associated with nanomolar level exposure of EDCs.

  13. X-ray Ne/O Ratio in Cataclysmic Variables

    Science.gov (United States)

    Schlegel, Eric M.; Rana, V.; Singh, K.; Girish, V.; Barrett, P.

    2006-12-01

    The Ne/O ratio has recently become a topic of considerable debate given the difficulties and discrepancies in recent measures of the Ne abundance in the Sun. We describe the results of an analysis of the permitted branches of the X-ray triplet transitions of Ne and O using the available spectra from the Chandra X-ray Observatory's High Energy Transmission Grating observations of cataclysmic variables. We compare our results with those obtained from the Sun and nearby stars.

  14. The hyperfine field on 19Ne in Fe

    International Nuclear Information System (INIS)

    Ekwall, B.; Johansson, K.; Lidbjoerk, P.; Lindgren, B.; Bedi, S.

    1984-04-01

    The hyperfine field on 19 Ne in iron was measured with the TDPAD method. At 77 K the 19 Ne hyperfine field in iron was found to be -4.48(7) T. The temperature dependence of the hyperfine field in iron was studied between 77 K and 650 K. It was not possible to observe any hyperfine field on 19 Ne in nickel, using the same method. (author)

  15. L'hydrogène Hydrogen

    Directory of Open Access Journals (Sweden)

    Balaceanu J. C.

    2006-11-01

    Full Text Available La crise pétrolière et le bouleversement du classement économique des énergies primaires qu'elle entraîne conjuguent leurs effets avec ceux d'une sensibilisation de l'opinion au respect de l'environnement pour favoriser l'avènement industriel d'innovations scientifiques et techniques dont l'intervention n'était prévisible que dans un avenir de plusieurs décennies. Le développement de l'énergie électrique nucléaire, qui actuellement s'impose économiquement, implique, pour élargir la pénétration de cette forme d'énergie à toutes les utilisations, une énergie chimique relais permettant un stockage et une régulation de la production; l'hydro- gène obtenu par électrolyse de l'eau semble pouvoir constituer ce combustible relais dans un délai raisonnable en tenant compte des contraintes de pollution. La chaleur nucléaire soulève a fortiori des problèmes identiques, elle peut théoriquement par dissociation thermique étagée de l'eau liquide fournir de l'hydrogène avec des rendements très satisfaisants, mais les problèmes de principe et de technologie posés par la mise en opération d'une suite de transformations chimiques et de séparations impliquant des composés particulièrement réactifs sont ardus et leur inventaire même n'est pas achevé. L'hydrogène, nouveau combustible polyvalent d'une industrie gazière perpétuelle, semble pouvoir bénéficier également, au niveau de son utilisation disséminée, de techniques nouvelles : stockages solides, turbines à hauts rendements, piles à combustible, qui ouvrent le marché de la traction et le marché électrique des installations isolées. Agent de réduction réactif et puissant, l'hydrogène peut également se substituer aux réducteurs conventionnels en métallurgie et donner une dimension nouvelle à l'hydrogénométallurgie par voie sèche ou par voie humide. Mais plus encore la mise en valeur économique des combustibles fossiles abondants . charbon, schistes

  16. Irène Jacob visits CERN

    CERN Multimedia

    CERN Bulletin

    2010-01-01

    French actress Irène Jacob, the daughter of physicist Maurice Jacob, visited the ATLAS and CMS control rooms on Monday 17 May together with Italian theatre actor-director Pippo Delbono, in search of inspiration for a short film. The film will be screened at the “nuit des particules” event accompanying this year’s ICHEP.   Pippo Delbono et Irène Jacob discussing their project. “La nuit des particules” (night of the particles) is an event open to the general public that is being organised for the evening of Tuesday, 27 July, to accompany the 35th International Conference on High Energy Physics (ICHEP). ICHEP is a major highlight in every physicist’s calendar, and this year’s edition is being held in Paris from 22 to 28 July. The short film will be screened during the evening, which will include a lecture and a show at the legendary Parisian cinema Le Grand Rex, with a colossal seating capacity of 2 700 spe...

  17. Lääne-Viru metskonnad osalevad pilootprojektis / Eva Klaas

    Index Scriptorium Estoniae

    Klaas, Eva, 1977-

    2008-01-01

    Riigimetsa majandamise keskuse Kirde regioonis alustati pilootprojektiga, mis seitsme metskonna territooriumil lahutab metsakasutuse ja -kasvatuse. Projektis osalevad Lääne-Virumaalt Triigi, Tudu ja Paasvere metskond

  18. Phase transition and angular momentum dependence of correlations in the rotational spectra of Ne20 and Ne22

    International Nuclear Information System (INIS)

    Satpathy, L.; Schmid, K.W.; Krewald, S.; Faessler, A.

    1974-01-01

    Multi-Configuration-Hartree-Fock (MCHF) calculations with angular momentum projection before the variation of the internal degree of freedom have been performed for the nuclei Ne 20 and Ne 22 . This procedure yields different correlated intrinsic states for the different members of a rotational band. Thus, the angular momentum dependence of correlations has been studied. Experimentally, the ground state spectra of Ne 20 and Ne 22 show properties similar to the phase transitions observed in some rare earth nuclei which have been well reproduced through the present calculations. The calculated spectra show a significant improvement compared to the ones obtained by variation before the angular momentum projection is effected. (author)

  19. 1:100,000 Grid of Louisiana, Geographic NAD83, LOSCO (1999)[quad100K_LOSCO_1999

    Data.gov (United States)

    Louisiana Geographic Information Center — This is a polygon dataset depicting the bounds of a regular 100K (60 minutes east-west by 30 minutes north-south) grid in geographic coordinates, NAD83, for the...

  20. A Novel Type II NAD+-Specific Isocitrate Dehydrogenase from the Marine Bacterium Congregibacter litoralis KT71.

    Directory of Open Access Journals (Sweden)

    Ming-Cai Wu

    Full Text Available In most living organisms, isocitrate dehydrogenases (IDHs convert isocitrate into ɑ-ketoglutarate (ɑ-KG. Phylogenetic analyses divide the IDH protein family into two subgroups: types I and II. Based on cofactor usage, IDHs are either NAD+-specific (NAD-IDH or NADP+-specific (NADP-IDH; NADP-IDH evolved from NAD-IDH. Type I IDHs include NAD-IDHs and NADP-IDHs; however, no type II NAD-IDHs have been reported to date. This study reports a novel type II NAD-IDH from the marine bacterium Congregibacter litoralis KT71 (ClIDH, GenBank accession no. EAQ96042. His-tagged recombinant ClIDH was produced in Escherichia coli and purified; the recombinant enzyme was NAD+-specific and showed no detectable activity with NADP+. The Km values of the enzyme for NAD+ were 262.6±7.4 μM or 309.1±11.2 μM with Mg2+ or Mn2+ as the divalent cation, respectively. The coenzyme specificity of a ClIDH Asp487Arg/Leu488His mutant was altered, and the preference of the mutant for NADP+ was approximately 24-fold higher than that for NAD+, suggesting that ClIDH is an NAD+-specific ancestral enzyme in the type II IDH subgroup. Gel filtration and analytical ultracentrifugation analyses revealed the homohexameric structure of ClIDH, which is the first IDH hexamer discovered thus far. A 163-amino acid segment of CIIDH is essential to maintain its polymerization structure and activity, as a truncated version lacking this region forms a non-functional monomer. ClIDH was dependent on divalent cations, the most effective being Mn2+. The maximal activity of purified recombinant ClIDH was achieved at 35°C and pH 7.5, and a heat inactivation experiment showed that a 20-min incubation at 33°C caused a 50% loss of ClIDH activity. The discovery of a NAD+-specific, type II IDH fills a gap in the current classification of IDHs, and sheds light on the evolution of type II IDHs.

  1. Investigations of the metabolism of NAD in embryonic neural tissue of mice after irradiation with X-rays

    International Nuclear Information System (INIS)

    Beuningen, M. van.

    1974-01-01

    Female mice of an institutes own inbred strain were killed on the 9th-13th day of pregnancy and the embryos were removed by caesarian section. The NAD content and protein content in the embryonic neural tissue of the mice increase the most from the 10th to 11th day. There is a relationship between NAD quantity and increase in size measured by the protein content. The enzymal activity of the NMN pyrophosphorylase runs parallel to the NAD rise and fall except for on the 11th day on which the enzyme increases further. The NAD biosynthesis from nicotinamide measured by the incorporation of 14-C nicotinamide in the NAD rises from the 10th to the 13th day. If one refers the incorporation to the protein content, however, the NAD synthesis falls from the 10th day onwards. An increase of the NAD content in the embryonic brain by the addition of nicotinamide in a high dose was not possible on the 10th and 12th day, whereas a clear increase was registered in the mother animal liver. Following an X-radiation with 200 R on the 9th day, the NAD content/brain dropped on the 11th day to its lowest point and had reached its normal value again on the 13th day, contrary to the protein content which only decreases on the 11th day. If one refers the NAD content, however, to protein quantity, then this only falls on the 10th day and rises on the 11th day almost to the normal value and has reached the latter by the 12th day. The NMN pyrophosphorylase activity falls on the 10th and 11th day, has its normal value on the 12th day and exceeds it on the 13th day. If one refers the enzyme activity to protein content, then it drops on the 10th day, reaches its lowest value on the 11th day, has its normal value on the 12th day and shoots above it on the 13th day. On the 10th day, the NAD content falls only after an X-ray with 200 R given on the 9th day, whereas the protein content remains constant. The NAD content does not change in the region of 50 to 150 R. (orig./LH) [de

  2. Effects of Kynurenine Pathway Metabolites on Intracellular NAD+ Synthesis and Cell Death in Human Primary Astrocytes and Neurons

    Directory of Open Access Journals (Sweden)

    Nady Braidy

    2009-01-01

    Full Text Available The kynurenine pathway (KP is a major route of L-tryptophan catabolism resulting in the production of the essential pyridine nucleotide nicotinamide adenine dinucleotide, (NAD+. Up-regulation of the KP during inflammation leads to the release of a number of biologically active metabolites into the brain. We hypothesised that while some of the extracellular KP metabolites may be beneficial for intracellular NAD+ synthesis and cell survival at physiological concentrations, they may contribute to neuronal and astroglial dysfunction and cell death at pathophysiological concentrations. In this study, we found that treatment of human primary neurons and astrocytes with 3-hydroxyanthranilic acid (3-HAA, 3-hydroxykynurenine (3-HK, quinolinic acid (QUIN, and picolinic acid (PIC at concentrations below 100 nM significantly increased intracellular NAD+ levels compared to non-treated cells. However, a dose dependent decrease in intracellular NAD+ levels and increased extracellular LDH activity was observed in human astrocytes and neurons treated with 3-HAA, 3-HK, QUIN and PIC at concentrations 100 nM and kynurenine (KYN, at concentrations above 1 μM. Intracellular NAD+ levels were unchanged in the presence of the neuroprotectant, kynurenic acid (KYNA, and a dose dependent increase in intracellular NAD+ levels was observed for TRP up to 1 mM. While anthranilic acid (AA increased intracellular NAD+ levels at concentration below 10 μM in astrocytes. NAD+ depletion and cell death was observed in AA treated neurons at concentrations above 500 nM. Therefore, the differing responses of astrocytes and neurons to an increase in KP metabolites should be considered when assessing KP toxicity during neuroinflammation.

  3. Effects of Kynurenine Pathway Metabolites on Intracellular NAD Synthesis and Cell Death in Human Primary Astrocytes and Neurons

    Directory of Open Access Journals (Sweden)

    Nady Braidy

    2009-01-01

    Full Text Available The kynurenine pathway (KP is a major route of L-tryptophan catabolism resulting in the production of the essential pyridine nucleotide nicotinamide adenine dinucleotide, (NAD + . Up-regulation of the KP during inflammation leads to the release of a number of biologically active metabolites into the brain. We hypothesised that while some of the extracellular KP metabolites may be beneficial for intracellular NAD + synthesis and cell survival at physiological concentrations, they may contribute to neuronal and astroglial dysfunction and cell death at pathophysiological concentrations. In this study, we found that treatment of human primary neurons and astrocytes with 3-hydroxyanthranilic acid (3-HAA, 3-hydroxykynurenine (3-HK, quinolinic acid (QUIN, and picolinic acid (PIC at concentrations below 100 nM significantly increased intracellular NAD + levels compared to non-treated cells. However, a dose dependent decrease in intracellular NAD + levels and increased extracellular LDH activity was observed in human astrocytes and neurons treated with 3-HAA, 3-HK, QUIN and PIC at concentrations >100 nM and kynurenine (KYN, at concentrations above 1 μM. Intracellular NAD + levels were unchanged in the presence of the neuroprotectant, kynurenic acid (KYNA, and a dose dependent increase in intracellular NAD + levels was observed for TRP up to 1 mM. While anthranilic acid (AA increased intracellular NAD + levels at concentration below 10 μM in astrocytes. NAD + depletion and cell death was observed in AA treated neurons at concentrations above 500 nM. Therefore, the differing responses of astrocytes and neurons to an increase in KP metabolites should be considered when assessing KP toxicity during neuroinflammation.

  4. Slowing ageing by design: the rise of NAD+ and sirtuin-activating compounds

    Science.gov (United States)

    Bonkowski, Michael S.; Sinclair, David A.

    2016-01-01

    The sirtuins (SIRT1–7) are a family of nicotinamide adenine dinucleotide (NAD+)-dependent deacylases with remarkable abilities to prevent diseases and even reverse aspects of ageing. Mice engineered to express additional copies of SIRT1 or SIRT6, or treated with sirtuin-activating compounds (STACs) such as resveratrol and SRT2104 or with NAD+ precursors, have improved organ function, physical endurance, disease resistance and longevity. Trials in non-human primates and in humans have indicated that STACs may be safe and effective in treating inflammatory and metabolic disorders, among others. These advances have demonstrated that it is possible to rationally design molecules that can alleviate multiple diseases and possibly extend lifespan in humans. PMID:27552971

  5. Flow-through 3D biofuel cell anode for NAD+-dependent enzymes

    International Nuclear Information System (INIS)

    Rincon, Rosalba A.; Lau, Carolin; Garcia, Kristen E.; Atanassov, Plamen

    2011-01-01

    NAD + -dependent enzymes require the presence of catalysts for cofactor regeneration in order to be employed in enzymatic biofuel cells. Poly-(methylene green) catalysts have proven to help the oxidation reaction of NADH allowing for the use of such enzymes in electrocatalytic oxidation reactions. In this paper we present the development of 3D anode based on NAD + -dependent malate dehydrogenase. The 3D material chosen was reticulated vitreous carbon (RVC) which was modified with poly-(MG) for NADH oxidation and it also accommodated the porous immobilization matrix for MDH consisting of MWCNTs embedded in chitosan; allowing for mass transport of the substrate to the electrode. Scanning electron microscopy was used in order to characterize the poly-(MG)-modified RVC, and electrochemical evaluation of the anode was performed.

  6. Calcium inhibition of the NAD+-linked isocitrate dehydrogenase from blowfly flight muscle mitochondria.

    Science.gov (United States)

    Bulos, B A; Thomas, B J; Sacktor, B

    1984-08-25

    Free Ca2+ was shown to inhibit the NAD+-isocitrate dehydrogenase from blowfly flight muscle mitochondria. Inhibition by free Ca2+ concentrations of 40 microM or greater was found in the absence or presence of ADP and citrate, two known activators of the enzyme. Calcium decreased the affinity of the enzyme for its substrate, the magnesium DL-isocitrate chelate; no change in the apparent V of the reaction was observed. Calcium was inhibitory when activity was measured in the presence of fixed concentrations of magnesium DL-isocitrate chelate in the presence of several fixed concentrations of either free isocitrate3-, an activator, or free Mg2+, an inhibitor of the enzyme. That NAD+-isocitrate dehydrogenase from blowfly flight muscle mitochondria was not activated by micromolar free Ca2+ is consistent with the view that calcium does not play a role in regulating the flux through the tricarboxylate cycle in this species.

  7. SIRT1-Mediated eNAMPT Secretion from Adipose Tissue Regulates Hypothalamic NAD+ and Function in Mice.

    Science.gov (United States)

    Yoon, Myeong Jin; Yoshida, Mitsukuni; Johnson, Sean; Takikawa, Akiko; Usui, Isao; Tobe, Kazuyuki; Nakagawa, Takashi; Yoshino, Jun; Imai, Shin-ichiro

    2015-05-05

    Nicotinamide phosphoribosyltransferase (NAMPT), the key NAD(+) biosynthetic enzyme, has two different forms, intra- and extracellular (iNAMPT and eNAMPT), in mammals. However, the significance of eNAMPT secretion remains unclear. Here we demonstrate that deacetylation of iNAMPT by the mammalian NAD(+)-dependent deacetylase SIRT1 predisposes the protein to secretion in adipocytes. NAMPT mutants reveal that SIRT1 deacetylates lysine 53 (K53) and enhances eNAMPT activity and secretion. Adipose tissue-specific Nampt knockout and knockin (ANKO and ANKI) mice show reciprocal changes in circulating eNAMPT, affecting hypothalamic NAD(+)/SIRT1 signaling and physical activity accordingly. The defect in physical activity observed in ANKO mice is ameliorated by nicotinamide mononucleotide (NMN). Furthermore, administration of a NAMPT-neutralizing antibody decreases hypothalamic NAD(+) production, and treating ex vivo hypothalamic explants with purified eNAMPT enhances NAD(+), SIRT1 activity, and neural activation. Thus, our findings indicate a critical role of adipose tissue as a modulator for the regulation of NAD(+) biosynthesis at a systemic level. Copyright © 2015 Elsevier Inc. All rights reserved.

  8. The NAD+ precursor nicotinamide riboside enhances oxidative metabolism and protects against high-fat diet induced obesity

    Science.gov (United States)

    Cantó, Carles; Houtkooper, Riekelt H.; Pirinen, Eija; Youn, Dou Y.; Oosterveer, Maaike H.; Cen, Yana; Fernandez-Marcos, Pablo J.; Yamamoto, Hiroyasu; Andreux, Pénélope A.; Cettour-Rose, Philippe; Gademann, Karl; Rinsch, Chris; Schoonjans, Kristina; Sauve, Anthony A.; Auwerx, Johan

    2013-01-01

    SUMMARY As NAD+ is a rate-limiting co-substrate for the sirtuin enzymes, its modulation is emerging as a valuable tool to regulate sirtuin function and, consequently, oxidative metabolism. In line with this premise, decreased activity of PARP-1 or CD38 —both NAD+ consumers— increases NAD+ bioavailability, resulting in SIRT1 activation and protection against metabolic disease. Here we evaluated whether similar effects could be achieved by increasing the supply of nicotinamide riboside (NR), a recently described natural NAD+ precursor with the ability to increase NAD+ levels, Sir2-dependent gene silencing and replicative lifespan in yeast. We show that NR supplementation in mammalian cells and mouse tissues increases NAD+ levels and activates SIRT1 and SIRT3, culminating in enhanced oxidative metabolism and protection against high fat diet-induced metabolic abnormalities. Consequently, our results indicate that the natural vitamin, NR, could be used as a nutritional supplement to ameliorate metabolic and age-related disorders characterized by defective mitochondrial function. PMID:22682224

  9. CD73 protein as a source of extracellular precursors for sustained NAD+ biosynthesis in FK866-treated tumor cells.

    Science.gov (United States)

    Grozio, Alessia; Sociali, Giovanna; Sturla, Laura; Caffa, Irene; Soncini, Debora; Salis, Annalisa; Raffaelli, Nadia; De Flora, Antonio; Nencioni, Alessio; Bruzzone, Santina

    2013-09-06

    NAD(+) is mainly synthesized in human cells via the "salvage" pathways starting from nicotinamide, nicotinic acid, or nicotinamide riboside (NR). The inhibition with FK866 of the enzyme nicotinamide phosphoribosyltransferase (NAMPT), catalyzing the first reaction in the "salvage" pathway from nicotinamide, showed potent antitumor activity in several preclinical models of solid and hematologic cancers. In the clinical studies performed with FK866, however, no tumor remission was observed. Here we demonstrate that low micromolar concentrations of extracellular NAD(+) or NAD(+) precursors, nicotinamide mononucleotide (NMN) and NR, can reverse the FK866-induced cell death, this representing a plausible explanation for the failure of NAMPT inhibition as an anti-cancer therapy. NMN is a substrate of both ectoenzymes CD38 and CD73, with generation of NAM and NR, respectively. In this study, we investigated the roles of CD38 and CD73 in providing ectocellular NAD(+) precursors for NAD(+) biosynthesis and in modulating cell susceptibility to FK866. By specifically silencing or overexpressing CD38 and CD73, we demonstrated that endogenous CD73 enables, whereas CD38 impairs, the conversion of extracellular NMN to NR as a precursor for intracellular NAD(+) biosynthesis in human cells. Moreover, cell viability in FK866-treated cells supplemented with extracellular NMN was strongly reduced in tumor cells, upon pharmacological inhibition or specific down-regulation of CD73. Thus, our study suggests that genetic or pharmacologic interventions interfering with CD73 activity may prove useful to increase cancer cell sensitivity to NAMPT inhibitors.

  10. The NAD(+) precursor nicotinamide riboside enhances oxidative metabolism and protects against high-fat diet-induced obesity.

    Science.gov (United States)

    Cantó, Carles; Houtkooper, Riekelt H; Pirinen, Eija; Youn, Dou Y; Oosterveer, Maaike H; Cen, Yana; Fernandez-Marcos, Pablo J; Yamamoto, Hiroyasu; Andreux, Pénélope A; Cettour-Rose, Philippe; Gademann, Karl; Rinsch, Chris; Schoonjans, Kristina; Sauve, Anthony A; Auwerx, Johan

    2012-06-06

    As NAD(+) is a rate-limiting cosubstrate for the sirtuin enzymes, its modulation is emerging as a valuable tool to regulate sirtuin function and, consequently, oxidative metabolism. In line with this premise, decreased activity of PARP-1 or CD38-both NAD(+) consumers-increases NAD(+) bioavailability, resulting in SIRT1 activation and protection against metabolic disease. Here we evaluated whether similar effects could be achieved by increasing the supply of nicotinamide riboside (NR), a recently described natural NAD(+) precursor with the ability to increase NAD(+) levels, Sir2-dependent gene silencing, and replicative life span in yeast. We show that NR supplementation in mammalian cells and mouse tissues increases NAD(+) levels and activates SIRT1 and SIRT3, culminating in enhanced oxidative metabolism and protection against high-fat diet-induced metabolic abnormalities. Consequently, our results indicate that the natural vitamin NR could be used as a nutritional supplement to ameliorate metabolic and age-related disorders characterized by defective mitochondrial function. Copyright © 2012 Elsevier Inc. All rights reserved.

  11. NAD-preferring malic enzyme: localization, regulation and its potential role in herring (Clupea harengus) sperm cells.

    Science.gov (United States)

    Niedźwiecka, Natalia; Gronczewska, Jadwiga; Skorkowski, Edward F

    2017-04-01

    Herring spermatozoa exhibit a high activity of NAD-preferring malic enzyme (NAD-ME). This enzyme is involved in the generation of NADH or NADPH in the decarboxylation of malate to form pyruvate and requires some divalent cations to express its activity. In order to confirm that NAD-ME isolated from herring sperm cells is localized in mitochondria, we performed immunofluorescent analysis and assayed spectrophotometrically the malic enzyme reaction. Production of polyclonal rabbit antibodies against NAD-ME from herring spermatozoa enabled identification of mitochondrial localization of this enzyme inside herring spermatozoa. The kinetic studies revealed that NAD-ME was competitively inhibited by ATP up to tenfold. Addition of fumarate reversed ATP-dependent inhibition of NAD-ME to 55 % of its maximum activity. The pH-dependent regulation of malic enzyme activity was also examined. Malic enzyme showed maximum activity at pH near 7.0 in all studied conditions. Finally, the role of malic enzyme activity regulation in mitochondria of herring sperm cells was discussed.

  12. Purification and properties of NAD(P)H: (quinone-acceptor) oxidoreductase of sugarbeet cells.

    Science.gov (United States)

    Trost, P; Bonora, P; Scagliarini, S; Pupillo, P

    1995-12-01

    NAD(P)H:(quinone-acceptor) oxidoreductase [NAD(P)H-QR], a plant cytosolic protein, was purified from cultured sugarbeet cells by a combination of ammonium sulfate fractionation, FPLC Superdex 200 gel filtration, Q-Sepharose anion-exchange chromatography, and a final Blue Sepharose CL-6B affinity chromatography with an NADPH gradient. The subunit molecular mass is 24 kDa and the active protein (94 kDa) is a tetramer. The isoelectric point is 4.9. The enzyme was characterized by ping-pong kinetics and extremely elevated catalytic capacity. It prefers NADPH over NADH as electron donor (kcat/Km ratios of 1.7 x 10(8) M-1 S-1 and 8.3 x 10(7) M-1 S-1 for NADPH and NADH, respectively, with benzoquinone as electron acceptor). The acridone derivative 7-iodo-acridone-4-carboxylic acid is an efficient inhibitor (I0.5 = 5 x 10(-5) M), dicumarol is weakly inhibitory. The best acceptor substances are hydrophilic, short-chain quinones such as ubiquinone-0 (Q-0), benzoquinone and menadione, followed by duroquinone and ferricyanide, whereas hydrophobic quinones, cytochrome c and oxygen are reduced at negligible rates at best. Quinone acceptors are reduced by a two-electron reaction with no apparent release of free semiquinonic intermediates. This and the above properties suggest some relationship of NAD(P)H-QR to DT-diaphorase, an animal flavoprotein which, however, has distinct structural properties and is strongly inhibited by dicumarol. It is proposed that NAD(P)H-QR by scavenging unreduced quinones and making them prone to conjugation may act in plant tissues as a functional equivalent of DT-diaphorase.

  13. Complementation of mitochondrial electron transport chain by manipulation of the NAD+/NADH ratio.

    Science.gov (United States)

    Titov, Denis V; Cracan, Valentin; Goodman, Russell P; Peng, Jun; Grabarek, Zenon; Mootha, Vamsi K

    2016-04-08

    A decline in electron transport chain (ETC) activity is associated with many human diseases. Although diminished mitochondrial adenosine triphosphate production is recognized as a source of pathology, the contribution of the associated reduction in the ratio of the amount of oxidized nicotinamide adenine dinucleotide (NAD(+)) to that of its reduced form (NADH) is less clear. We used a water-forming NADH oxidase from Lactobacillus brevis (LbNOX) as a genetic tool for inducing a compartment-specific increase of the NAD(+)/NADH ratio in human cells. We used LbNOX to demonstrate the dependence of key metabolic fluxes, gluconeogenesis, and signaling on the cytosolic or mitochondrial NAD(+)/NADH ratios. Expression of LbNOX in the cytosol or mitochondria ameliorated proliferative and metabolic defects caused by an impaired ETC. The results underscore the role of reductive stress in mitochondrial pathogenesis and demonstrate the utility of targeted LbNOX for direct, compartment-specific manipulation of redox state. Copyright © 2016, American Association for the Advancement of Science.

  14. The chemistry of nicotinamide adenine dinucleotide (NAD) analogues containing C-nucleosides related to nicotinamide riboside.

    Science.gov (United States)

    Pankiewicz, Krzysztof W; Watanabe, Kyoichi A; Lesiak-Watanabe, Krystyna; Goldstein, Barry M; Jayaram, Hiremagalur N

    2002-04-01

    Oncolytic C-nucleosides, tiazofurin (2-beta-D-ribofuranosylthiazole-4-carboxamide) and benzamide riboside (3-beta-D-ribofuranosylbenzamide) are converted in cell into active metabolites thiazole-4-carboxamide- and benzamide adenine dinucleotide, TAD and BAD, respectively. TAD and BAD as NAD analogues were found to bind at the nicotinamide adenine dinucleotide (cofactor NAD) site of inosine monophosphate dehydrogenase (IMPDH), an important target in cancer treatment. The synthesis and evaluation of anticancer activity of a number of C-nucleosides related to tiazofurin and nicotinamide riboside then followed and are reviewed herein. Interestingly, pyridine C-nucleosides (such as C-nicotinamide riboside) are not metabolized into the corresponding NAD analogues in cell. Their conversion by chemical methods is described. As dinucleotides these compounds show inhibition of IMPDH in low micromolar level. Also, the synthesis of BAD in metabolically stable bis(phosphonate) form is discussed indicating the usefulness of such preformed inhibitors in drug development. Among tiazofurin analogues, Franchetti and Grifantini found, that the replacement of the sulfur by oxygen (as in oxazafurin) but not the removal of nitrogen (tiophenfurin) of the thiazole ring resulted in inactive compounds. The anti cancer activity of their synthetic dinucleotide analogues indicate that inactive compounds are not only poorly metabolized in cell but also are weak inhibitors of IMPDH as dinucleotides.

  15. Melatonin protects chondrocytes from impairment induced by glucocorticoids via NAD+-dependent SIRT1.

    Science.gov (United States)

    Yang, Wei; Kang, Xiaomin; Qin, Na; Li, Feng; Jin, Xinxin; Ma, Zhengmin; Qian, Zhuang; Wu, Shufang

    2017-10-01

    Intra-articular injection of glucocorticoids is used to relieve pain and inflammation in osteoarthritis patients, which is occasionally accompanied with the serious side effects of glucocorticoids in collagen-producing tissue. Melatonin is the major hormone released from the pineal gland and its beneficial effects on cartilage has been suggested. In the present study, we investigated the protective role of melatonin on matrix degeneration in chondrocytes induced by dexamethasone (Dex). The chondrocytes isolated from mice knee joint were treated with Dex, melatonin, EX527 and siRNA targeted for SIRT6, respectively. Dex treatment induced the loss of the extracellular matrix, NAD + /NADH ratio and NADPH concentration in chondrocytes. Melatonin alone have no effect on the quantity of proteoglycans and collagen type IIa1, however, the pretreatment of melatonin reversed the negative effects induced by Dex. Meanwhile, the significant decrease in NAD + /NADH ratio and NADPH concentration in Dex group were up-regulated by pretreatment of melatonin. Furthermore, it was revealed that inhibition of SIRT1 blocked the protective effects of melatonin. The enhancement of NAD + -dependent SIRT1 activity contributes to the chondroprotecfive effects of melatonin, which has a great benefit to prevent dexamethasone-induced chondrocytes impairment. Copyright © 2017 Elsevier Inc. All rights reserved.

  16. The hidden life of NAD+-consuming ectoenzymes in the endocrine system.

    Science.gov (United States)

    Malavasi, Fabio; Deaglio, Silvia; Zaccarello, Gianluca; Horenstein, Alberto L; Chillemi, Antonella; Audrito, Valentina; Serra, Sara; Gandione, Marina; Zitella, Andrea; Tizzani, Alessandro

    2010-10-01

    Ectoenzymes are a family of cell surface molecules whose catalytic domain lies in the extracellular region. A subset of this family, nucleotide-metabolizing ectoenzymes, are key components in the regulation of the extracellular balance between nucleotides (e.g. NAD(+) or ATP) and nucleosides (e.g. adenosine). Their substrates and products are signalling molecules that act by binding to specific receptors, triggering signals that regulate a variety of functions, ranging from the migration of immune cells, to synaptic transmission in the brain, to hormone/receptor interactions in the glands. Almost two decades of accumulated data indicate that these regulatory processes significantly affect the endocrine system, a tightly controlled information signal complex with clear evidence of fine regulation. Functional models discussed in this review include insulin secretion, bone modelling and the association between hormones and behaviour. The emerging pattern is one of a system operating as a scale-free network that hinges around hubs of key molecules, such as NAD(+) or ATP. The underlying natural link between nucleotides, ectoenzymes and the endocrine system is far from being clearly demonstrated. However, the body of evidence supporting the existence of such connection is growing exponentially. This review will try to read the available evidence in a hypothesis-oriented perspective, starting from the description of NAD(+) and of ecto- and endoenzymes involved in its metabolism.

  17. ARTD1/PARP1 Negatively Regulates Glycolysis by Inhibiting Hexokinase 1 Independent of NAD+ Depletion

    Directory of Open Access Journals (Sweden)

    Elise Fouquerel

    2014-09-01

    Full Text Available ARTD1 (PARP1 is a key enzyme involved in DNA repair through the synthesis of poly(ADP-ribose (PAR in response to strand breaks, and it plays an important role in cell death following excessive DNA damage. ARTD1-induced cell death is associated with NAD+ depletion and ATP loss; however, the molecular mechanism of ARTD1-mediated energy collapse remains elusive. Using real-time metabolic measurements, we compared the effects of ARTD1 activation and direct NAD+ depletion. We found that ARTD1-mediated PAR synthesis, but not direct NAD+ depletion, resulted in a block to glycolysis and ATP loss. We then established a proteomics-based PAR interactome after DNA damage and identified hexokinase 1 (HK1 as a PAR binding protein. HK1 activity is suppressed following nuclear ARTD1 activation and binding by PAR. These findings help explain how prolonged activation of ARTD1 triggers energy collapse and cell death, revealing insight into the importance of nucleus-to-mitochondria communication via ARTD1 activation.

  18. NH3-dependent NAD+ synthetase from Bacillus subtilis at 1 A resolution.

    Science.gov (United States)

    Symersky, Jindrich; Devedjiev, Yancho; Moore, Karen; Brouillette, Christie; DeLucas, Larry

    2002-07-01

    The final step of NAD+ biosynthesis includes an amide transfer to nicotinic acid adenine dinucleotide (NaAD) catalyzed by NAD+ synthetase. This enzyme was co-crystallized in microgravity with natural substrates NaAD and ATP at pH 8.5. The crystal was exposed to ammonium ions, synchrotron diffraction data were collected and the atomic model was refined anisotropically at 1 A resolution to R = 11.63%. Both binding sites are occupied by the NAD-adenylate intermediate, pyrophosphate and two magnesium ions. The atomic resolution of the structure allows better definition of non-planar peptide groups, reveals a low mean anisotropy of protein and substrate atoms and indicates the H-atom positions of the phosphoester group of the reaction intermediate. The phosphoester group is protonated at the carbonyl O atom O7N, suggesting a carbenium-ion structure stabilized by interactions with two solvent sites presumably occupied by ammonia and a water molecule. A mechanism is proposed for the second catalytic step, which includes a nucleophilic attack by the ammonia molecule on the intermediate.

  19. Degradation of (-)-ephedrine by Pseudomonas putida. Detection of (-)-ephedrine: NAD+-oxidoreductase from Arthrobacter globiformis.

    Science.gov (United States)

    Klamann, E; Lingens, F

    1980-01-01

    A bacterium utilizing the alkaloid (-)-ephedrine as its sole source of carbon was isolated by an enrichment-culture technique from soil supplemented with 4-benzoyl-1,3-oxazolidinon-(2). The bacterium was indentified as Pseudomonas putida by morphological and physiological studies. The following metabolites were isolated from the culture fluid: methylamine, formaldehyde, methylbenzoylcarbinol (2-hydroxy-1-oxo-1 phenylpropane), benzoid acid, pyrocatechol and cis, cis-muconic acid. A pathway for the degradation of (-)-ephedrine by Pseudomonas putida is proposed and compared with the degradative pathway in Arthrobacter globiformis. The enzyme, which is responsible for the first step in the catabolism of (-)-ephedrine could be demonstrated in extracts from Arthrobacter globiformis. This enzyme catalyses the dehydrogenation of (-)-ephedrine yielding phenylacetylcarbinol/methylbenzoylcarbinol and methylamine. It requires NAD+ as cofactor and exhibits optimal activity at pH 11 in 0.1 M glycine/NaOH buffer. The Km value for (-)-ephedrine is 0.02 mM and for NAD+ 0.11 mM, respectively. No remarkable loss of activity is observed following treatment with EDTA. The enzyme has been shown to react with a wide range of ethanolamines. A slight enrichment was obtained by ammonium sulphate precipitation. The name (-)-ephedrine: NAD+-oxidoreductase (deaminating) is proposed.

  20. Oleate ameliorates palmitate-induced reduction of NAMPT activity and NAD levels in primary human hepatocytes and hepatocarcinoma cells.

    Science.gov (United States)

    Penke, Melanie; Schuster, Susanne; Gorski, Theresa; Gebhardt, Rolf; Kiess, Wieland; Garten, Antje

    2017-10-03

    Nicotinamide phosphoribosyltransferase (NAMPT) and nicotinamide adenine dinucleotide (NAD) levels are crucial for liver function. The saturated fatty acid palmitate and the unsaturated fatty acid oleate are the main free fatty acids in adipose tissue and human diet. We asked how these fatty acids affect cell survival, NAMPT and NAD levels in HepG2 cells and primary human hepatocytes. HepG2 cells were stimulated with palmitate (0.5mM), oleate (1mM) or a combination of both (0.5mM/1mM) as well as nicotinamide mononucleotide (NMN) (0.5 mM) or the specific NAMPT inhibitor FK866 (10nM). Cell survival was measured by WST-1 assay and Annexin V/propidium iodide staining. NAD levels were determined by NAD/NADH Assay or HPLC. Protein and mRNA levels were analysed by Western blot analyses and qPCR, respectively. NAMPT enzyme activity was measured using radiolabelled 14 C-nicotinamide. Lipids were stained by Oil red O staining. Palmitate significantly reduced cell survival and induced apoptosis at physiological doses. NAMPT activity and NAD levels significantly declined after 48h of palmitate. In addition, NAMPT mRNA expression was enhanced which was associated with increased NAMPT release into the supernatant, while intracellular NAMPT protein levels remained stable. Oleate alone did not influence cell viability and NAMPT activity but ameliorated the negative impact of palmitate on cell survival, NAMPT activity and NAD levels, as well as the increased NAMPT mRNA expression and secretion. NMN was able to normalize intracellular NAD levels but did not ameliorate cell viability after co-stimulation with palmitate. FK866, a specific NAMPT inhibitor did not influence lipid accumulation after oleate-treatment. Palmitate targets NAMPT activity with a consequent cellular depletion of NAD. Oleate protects from palmitate-induced apoptosis and variation of NAMPT and NAD levels. Palmitate-induced cell stress leads to an increase of NAMPT mRNA and accumulation in the supernatant. However

  1. Nicotinamide riboside promotes Sir2 silencing and extends lifespan via Nrk and Urh1/Pnp1/Meu1 pathways to NAD+.

    Science.gov (United States)

    Belenky, Peter; Racette, Frances G; Bogan, Katrina L; McClure, Julie M; Smith, Jeffrey S; Brenner, Charles

    2007-05-04

    Although NAD(+) biosynthesis is required for Sir2 functions and replicative lifespan in yeast, alterations in NAD(+) precursors have been reported to accelerate aging but not to extend lifespan. In eukaryotes, nicotinamide riboside is a newly discovered NAD(+) precursor that is converted to nicotinamide mononucleotide by specific nicotinamide riboside kinases, Nrk1 and Nrk2. In this study, we discovered that exogenous nicotinamide riboside promotes Sir2-dependent repression of recombination, improves gene silencing, and extends lifespan without calorie restriction. The mechanism of action of nicotinamide riboside is totally dependent on increased net NAD(+) synthesis through two pathways, the Nrk1 pathway and the Urh1/Pnp1/Meu1 pathway, which is Nrk1 independent. Additionally, the two nicotinamide riboside salvage pathways contribute to NAD(+) metabolism in the absence of nicotinamide-riboside supplementation. Thus, like calorie restriction in the mouse, nicotinamide riboside elevates NAD(+) and increases Sir2 function.

  2. Investigations of the radiosensitivity of enzymes of the NAD metabolism localized in the cell nucleus in the spleen of white mice

    International Nuclear Information System (INIS)

    Beisel, P.

    1975-01-01

    The radiosensitivity of enzymes of the NAD metabolism localized in the cell nuclei and of NAD glycohydrolase in the total homogenate of the spleen of white mice was investigated. At the same time the DNA and protein contents were determined. After whole-body irradiation with 510 R, the activity of NAD pyrophosphorylase and NAD glycohydrolase located in the cell nuclei is markedly lower as early as 3 hours after irradiation; this decrease is noticeable until the 10th day after irradiation. With regard to the dose dependence of the radiosensitivity at 6 and 24 hours after irradiation, it was found that NAD pyrophosphorylase and the NAD glycohydrolase localized in the cell nuclei were very radiosensitive even at doses [de

  3. Nicotinamide Riboside and Nicotinic Acid Riboside Salvage in Fungi and Mammals: QUANTITATIVE BASIS FOR Urh1 AND PURINE NUCLEOSIDE PHOSPHORYLASE FUNCTION IN NAD+METABOLISM*S⃞

    OpenAIRE

    Belenky, Peter; Christensen, Kathryn C.; Gazzaniga, Francesca; Pletnev, Alexandre A.; Brenner, Charles

    2009-01-01

    NAD+ is a co-enzyme for hydride transfer enzymes and an essential substrate of ADP-ribose transfer enzymes and sirtuins, the type III protein lysine deacetylases related to yeast Sir2. Supplementation of yeast cells with nicotinamide riboside extends replicative lifespan and increases Sir2-dependent gene silencing by virtue of increasing net NAD+ synthesis. Nicotinamide riboside elevates NAD+ levels via the nicotinamide riboside kinase pathway and by a pathway initiate...

  4. Deep Learning MicroBooNE

    Science.gov (United States)

    Genty, Victor; Terao, Kazuhiro; Wonjirad, Taritree

    2017-01-01

    The Liquid Argon Time Projection Chamber (LAr TPC) technology provides a high resolution image of ionizing particle trajectories raising a need for new event reconstruction techniques based on pattern recognition. The traditional bottoms-up reconstruction approach to extract physics involves a complex sequence of signal waveform processing, 2D and/or 3D geometrical pattern recognition, calorimetry, and finally particle identification before a neutrino interaction can be identified in an event. We present a top-down reconstruction approach using a machine learning algorithm called Deep Learning which uses convolutional neural networks to find a neutrino interaction in a LAr TPC image. We trained our network on images of simulated single particles and neutrinos overlaid on cosmic-ray background data taken from the MicroBooNE detector. In this talk, we present our result that shows convolutional networks can successfully learn LAr TPC images to perform particle identification, neutrino event selection, and localization of a neutrino interaction vertex in a large LAr TPC image.

  5. Exclusive measurements of nuclear breakup reactions of 17Ne

    Directory of Open Access Journals (Sweden)

    Wamers F.

    2014-03-01

    Full Text Available We have studied one-proton-removal reactions of about 500MeV/u 17Ne beams on a carbon target at the R3B/LAND setup at GSI by detecting beam-like 15O-p and determining their relative-energy distribution. We exclusively selected the removal of a 17Ne halo proton, and the Glauber-model analysis of the 16F momentum distribution resulted in an s2 contribution in the 17Ne ground state of about 40%.

  6. NAD(+)-dependent activation of Sirt1 corrects the phenotype in a mouse model of mitochondrial disease.

    Science.gov (United States)

    Cerutti, Raffaele; Pirinen, Eija; Lamperti, Costanza; Marchet, Silvia; Sauve, Anthony A; Li, Wei; Leoni, Valerio; Schon, Eric A; Dantzer, Françoise; Auwerx, Johan; Viscomi, Carlo; Zeviani, Massimo

    2014-06-03

    Mitochondrial disorders are highly heterogeneous conditions characterized by defects of the mitochondrial respiratory chain. Pharmacological activation of mitochondrial biogenesis has been proposed as an effective means to correct the biochemical defects and ameliorate the clinical phenotype in these severely disabling, often fatal, disorders. Pathways related to mitochondrial biogenesis are targets of Sirtuin1, a NAD(+)-dependent protein deacetylase. As NAD(+) boosts the activity of Sirtuin1 and other sirtuins, intracellular levels of NAD(+) play a key role in the homeostatic control of mitochondrial function by the metabolic status of the cell. We show here that supplementation with nicotinamide riboside, a natural NAD(+) precursor, or reduction of NAD(+) consumption by inhibiting the poly(ADP-ribose) polymerases, leads to marked improvement of the respiratory chain defect and exercise intolerance of the Sco2 knockout/knockin mouse, a mitochondrial disease model characterized by impaired cytochrome c oxidase biogenesis. This strategy is potentially translatable into therapy of mitochondrial disorders in humans. Copyright © 2014 The Authors. Published by Elsevier Inc. All rights reserved.

  7. Bioinspired Design of Alcohol Dehydrogenase@nano TiO2 Microreactors for Sustainable Cycling of NAD+/NADH Coenzyme

    Directory of Open Access Journals (Sweden)

    Sen Lin

    2018-02-01

    Full Text Available The bioinspired design and construction of enzyme@capsule microreactors with specific cell-like functionality has generated tremendous interest in recent years. Inspired by their fascinating complexity, scientists have endeavored to understand the essential aspects of a natural cell and create biomimicking microreactors so as to immobilize enzymes within the hierarchical structure of a microcapsule. In this study, simultaneous encapsulation of alcohol dehydrogenase (ADH was achieved during the preparation of microcapsules by the Pickering emulsion method using amphiphilic modified TiO2 nanoparticles (NPs as building blocks for assembling the photocatalytic microcapsule membrane. The ADH@TiO2 NP microreactors exhibited dual catalytic functions, i.e., spatially confined enzymatic catalysis and the membrane-associated photocatalytic oxidation under visible light. The sustainable cycling of nicotinamide adenine dinucleotide (NAD coenzyme between NADH and NAD+ was realized by enzymatic regeneration of NADH from NAD+ reduction, and was provided in a form that enabled further photocatalytic oxidation to NAD+ under visible light. This bioinspired ADH@TiO2 NP microreactor allowed the linking of a semiconductor mineral-based inorganic photosystem to enzymatic reactions. This is a first step toward the realization of sustainable biological cycling of NAD+/NADH coenzyme in synthetic functional microsystems operating under visible light irradiation.

  8. Converting NADH to NAD+ by nicotinamide nucleotide transhydrogenase as a novel strategy against mitochondrial pathologies during aging.

    Science.gov (United States)

    Olgun, Abdullah

    2009-08-01

    Mitochondrial DNA defects are involved supposedly via free radicals in many pathologies including aging and cancer. But, interestingly, free radical production was not found increased in prematurely aging mice having higher mutation rate in mtDNA. Therefore, some other mechanisms like the increase of mitochondrial NADH/NAD(+) and ubiquinol/ubiquinone ratios, can be in action in respiratory chain defects. NADH/NAD(+) ratio can be normalized by the activation or overexpression of nicotinamide nucleotide transhydrogenase (NNT), a mitochondrial enzyme catalyzing the following very important reaction: NADH + NADP(+ ) NADPH + NAD(+). The products NAD(+) and NADPH are required in many critical biological processes, e.g., NAD(+) is used by histone deacetylase Sir2 which regulates longevity in different species. NADPH is used in a number of biosynthesis reactions (e.g., reduced glutathione synthesis), and processes like apoptosis. Increased ubiquinol/ubiquinone ratio interferes the function of dihydroorotate dehydrogenase, the only mitochondrial enzyme involved in ubiquinone mediated de novo pyrimidine synthesis. Uridine and its prodrug triacetyluridine are used to compensate pyrimidine deficiency but their bioavailability is limited. Therefore, the normalization of the ubiquinol/ubiquinone ratio can be accomplished by allotopic expression of alternative oxidase, a mitochondrial ubiquinol oxidase which converts ubiquinol to ubiquinone.

  9. Coulomb and nuclear excitations of narrow resonances in 17Ne

    Directory of Open Access Journals (Sweden)

    J. Marganiec

    2016-08-01

    Full Text Available New experimental data for dissociation of relativistic 17Ne projectiles incident on targets of lead, carbon, and polyethylene targets at GSI are presented. Special attention is paid to the excitation and decay of narrow resonant states in 17Ne. Distributions of internal energy in the O15+p+p three-body system have been determined together with angular and partial-energy correlations between the decay products in different energy regions. The analysis was done using existing experimental data on 17Ne and its mirror nucleus 17N. The isobaric multiplet mass equation is used for assignment of observed resonances and their spins and parities. A combination of data from the heavy and light targets yielded cross sections and transition probabilities for the Coulomb excitations of the narrow resonant states. The resulting transition probabilities provide information relevant for a better understanding of the 17Ne structure.

  10. Sensitivity studies for the KM3NeT

    Energy Technology Data Exchange (ETDEWEB)

    Shanidze, R.; Kuch, S. [Physics Inst. 1, Univ. of Erlangen-Nuremberg (Germany)

    2007-07-01

    KM3NeT is an European deep-sea research infrastructure, which will host a neutrino telescope with a volume of at least one cubic kilometre at the bottom of the Mediterranean Sea. The parameters and technical specifications of KM3NeT project will be finalized in the framework of EU-funded Design Study which started in 2006. KM3NeT neutrino telescope will search for the point sources of extra-terrestrial high energy neutrinos as well as for diffuse flux from unidentified sources. The sensitivity for these neutrino fluxes have been studied for the different models of KM3NeT detector, including various geometries and optical modules. The preliminary results of these studies, obtained with Monte Carlo simulations are presented in the talk. (orig.)

  11. Simulation studies for KM3NeT

    Energy Technology Data Exchange (ETDEWEB)

    Shanidze, Rezo [Erlangen Centre for Astroparticle Physics (ECAP), Erlangen University, Erwin-Rommel-Str. 1, 91058 Erlangen (Germany)

    2009-07-01

    KM3NeT is a future European deep-sea research infrastructure in the Mediterranean Sea, which will host the world's most sensitive high-energy neutrino telescope. The Design Study for the KM3NeT infrastructure is supported by the EU in FP6 and started in 2006. The KM3NeT consortium, which is formed by the ANTARES, NEMO and NESTOR collaborations as well as marine science and technology institutes, is currently investigating two design options for the final document of the Design Study, the KM3NeT technical design report (TDR). These options and the results of corresponding Monte Carlo studies are presented and discussed.

  12. Discharge modulation noise in He---Ne laser radiation

    NARCIS (Netherlands)

    Bolwijn, P.T.

    1967-01-01

    Discharge modulation noise in He---Ne laser radiation is considered theoretically, including explicitly the laser oscillator properties. Experiments reported previously by us and other authors are in agreement with our analysis.

  13. WE FRIENDS, Lääne-Eesti arengupartnerlus / Ingrit Kera

    Index Scriptorium Estoniae

    Kera, Ingrit

    2006-01-01

    Naised saavad osa hiidlaste kirjutatud europrojektist "We Friends", mille eesmärk on Lääne-Eesti madala konkurentsivõimega naiste ja lapsi üksi kasvatavate noorte emade tööhõivele kaasaaitamine

  14. neXtProt: a knowledge platform for human proteins.

    Science.gov (United States)

    Lane, Lydie; Argoud-Puy, Ghislaine; Britan, Aurore; Cusin, Isabelle; Duek, Paula D; Evalet, Olivier; Gateau, Alain; Gaudet, Pascale; Gleizes, Anne; Masselot, Alexandre; Zwahlen, Catherine; Bairoch, Amos

    2012-01-01

    neXtProt (http://www.nextprot.org/) is a new human protein-centric knowledge platform. Developed at the Swiss Institute of Bioinformatics (SIB), it aims to help researchers answer questions relevant to human proteins. To achieve this goal, neXtProt is built on a corpus containing both curated knowledge originating from the UniProtKB/Swiss-Prot knowledgebase and carefully selected and filtered high-throughput data pertinent to human proteins. This article presents an overview of the database and the data integration process. We also lay out the key future directions of neXtProt that we consider the necessary steps to make neXtProt the one-stop-shop for all research projects focusing on human proteins.

  15. 77 FR 6481 - Television Broadcasting Services; Lincoln, NE

    Science.gov (United States)

    2012-02-08

    ...] Television Broadcasting Services; Lincoln, NE AGENCY: Federal Communications Commission. ACTION: Final rule... power television rulemaking petitions requesting channel substitutions in May 2011, it subsequently... CFR Part 73 Television. Federal Communications Commission. Barbara A. Kreisman, Chief, Video Division...

  16. Investigation of 35S NE-78241 mobility in plants

    International Nuclear Information System (INIS)

    Enisz, J.; Orsos, S.

    1982-01-01

    The mobility of 35 S NE-78241 (N-iso-thiocyanato-methyl-2,6-dimethyl-chloracetanilide) in plants has been studied. The compound is not absorbed via the leaves from aqueous solutions. It shows active transport through the root-system. It is strongly bound to soil. In bean plant (Phaseolus vulgaris) inoculated with Uromyces appendiculatus 35 S NE-78241 is selectively enriched at the place of infection. (author)

  17. NESTOR participation in the KM3NeT

    CERN Document Server

    Anassontzis, E

    2008-01-01

    The NESTOR Collaboration is a leading participant in the Design Study of the KM3NeT, the European Deep Sea Neutrino Telescope. In this report we describe briefly the KM3NeT and the NESTOR experience and contribution towards this objective; the 4500m deep NESTOR site, the star-like detector, the deployment and recovery of telescope modules and the "DELTA-BERENIKE", the specially constructed deployment ship.

  18. The MicroBooNE Technical Design Report

    Energy Technology Data Exchange (ETDEWEB)

    Fleming, Bonnie [Fermi National Accelerator Lab. (FNAL), Batavia, IL (United States)

    2012-02-24

    MicroBooNE will build, operate, and extract physics from the first large liquid argon time projection chamber (LArTPC) that will be exposed to a high-intensity neutrino beam. With its unparalleled capabilities in tracking, vertexing, calorimetry, and particle identification, all with full electronic readout, MicroBooNE represents a major advance in detector technology for neutrino physics in the energy regime of most importance for elucidating oscillation phenomena.

  19. A Key Enzyme of the NAD+ Salvage Pathway in Thermus thermophilus: Characterization of Nicotinamidase and the Impact of Its Gene Deletion at High Temperatures.

    Science.gov (United States)

    Taniguchi, Hironori; Sungwallek, Sathidaphorn; Chotchuang, Phatcharin; Okano, Kenji; Honda, Kohsuke

    2017-09-01

    NAD (NAD + ) is a cofactor related to many cellular processes. This cofactor is known to be unstable, especially at high temperatures, where it chemically decomposes to nicotinamide and ADP-ribose. Bacteria, yeast, and higher organisms possess the salvage pathway for reconstructing NAD + from these decomposition products; however, the importance of the salvage pathway for survival is not well elucidated, except for in pathogens lacking the NAD + de novo synthesis pathway. Herein, we report the importance of the NAD + salvage pathway in the thermophilic bacterium Thermus thermophilus HB8 at high temperatures. We identified the gene encoding nicotinamidase (TTHA0328), which catalyzes the first reaction of the NAD + salvage pathway. This recombinant enzyme has a high catalytic activity against nicotinamide ( K m of 17 μM, k cat of 50 s -1 , k cat / K m of 3.0 × 10 3 s -1 · mM -1 ). Deletion of this gene abolished nicotinamide deamination activity in crude extracts of T. thermophilus and disrupted the NAD + salvage pathway in T. thermophilus Disruption of the salvage pathway led to the severe growth retardation at a higher temperature (80°C), owing to the drastic decrease in the intracellular concentrations of NAD + and NADH. IMPORTANCE NAD + and other nicotinamide cofactors are essential for cell metabolism. These molecules are unstable and decompose, even under the physiological conditions in most organisms. Thermophiles can survive at high temperatures where NAD + decomposition is, in general, more rapid. This study emphasizes that NAD + instability and its homeostasis can be one of the important factors for thermophile survival in extreme temperatures. Copyright © 2017 American Society for Microbiology.

  20. New function for Escherichia coli xanthosine phophorylase (xapA): genetic and biochemical evidences on its participation in NAD+ salvage from nicotinamide

    Science.gov (United States)

    2014-01-01

    Background In an effort to reconstitute the NAD+ synthetic pathway in Escherichia coli (E. coli), we produced a set of gene knockout mutants with deficiencies in previously well-defined NAD+de novo and salvage pathways. Unexpectedly, the mutant deficient in NAD+de novo and salvage pathway I could grow in M9/nicotinamide medium, which was contradictory to the proposed classic NAD+ metabolism of E. coli. Such E. coli mutagenesis assay suggested the presence of an undefined machinery to feed nicotinamide into the NAD+ biosynthesis. We wanted to verify whether xanthosine phophorylase (xapA) contributed to a new NAD+ salvage pathway from nicotinamide. Results Additional knockout of xapA further slowed down the bacterial growth in M9/nicotinamide medium, whereas the complementation of xapA restored the growth phenotype. To further validate the new function of xapA, we cloned and expressed E. coli xapA as a recombinant soluble protein. Biochemical assay confirmed that xapA was capable of using nicotinamide as a substrate for nicotinamide riboside formation. Conclusions Both the genetic and biochemical evidences indicated that xapA could convert nicotinamide to nicotinamide riboside in E. coli, albeit with relatively weak activity, indicating that xapA may contribute to a second NAD+ salvage pathway from nicotinamide. We speculate that this xapA-mediated NAD+ salvage pathway might be significant in some bacteria lacking NAD+de novo and NAD+ salvage pathway I or II, to not only use nicotinamide riboside, but also nicotinamide as precursors to synthesize NAD+. However, this speculation needs to be experimentally tested. PMID:24506841

  1. New function for Escherichia coli xanthosine phophorylase (xapA): genetic and biochemical evidences on its participation in NAD(+) salvage from nicotinamide.

    Science.gov (United States)

    Dong, Wei-Ren; Sun, Cen-Cen; Zhu, Guan; Hu, Shi-Hua; Xiang, Li-Xin; Shao, Jian-Zhong

    2014-02-08

    In an effort to reconstitute the NAD(+) synthetic pathway in Escherichia coli (E. coli), we produced a set of gene knockout mutants with deficiencies in previously well-defined NAD(+)de novo and salvage pathways. Unexpectedly, the mutant deficient in NAD(+) de novo and salvage pathway I could grow in M9/nicotinamide medium, which was contradictory to the proposed classic NAD(+) metabolism of E. coli. Such E. coli mutagenesis assay suggested the presence of an undefined machinery to feed nicotinamide into the NAD(+) biosynthesis. We wanted to verify whether xanthosine phophorylase (xapA) contributed to a new NAD(+) salvage pathway from nicotinamide. Additional knockout of xapA further slowed down the bacterial growth in M9/nicotinamide medium, whereas the complementation of xapA restored the growth phenotype. To further validate the new function of xapA, we cloned and expressed E. coli xapA as a recombinant soluble protein. Biochemical assay confirmed that xapA was capable of using nicotinamide as a substrate for nicotinamide riboside formation. Both the genetic and biochemical evidences indicated that xapA could convert nicotinamide to nicotinamide riboside in E. coli, albeit with relatively weak activity, indicating that xapA may contribute to a second NAD(+) salvage pathway from nicotinamide. We speculate that this xapA-mediated NAD(+) salvage pathway might be significant in some bacteria lacking NAD(+) de novo and NAD(+) salvage pathway I or II, to not only use nicotinamide riboside, but also nicotinamide as precursors to synthesize NAD(+). However, this speculation needs to be experimentally tested.

  2. Acute Ethanol Intake Induces NAD(PH Oxidase Activation and Rhoa Translocation in Resistance Arteries

    Directory of Open Access Journals (Sweden)

    Janaina A. Simplicio

    Full Text Available Abstract Background: The mechanism underlying the vascular dysfunction induced by ethanol is not totally understood. Identification of biochemical/molecular mechanisms that could explain such effects is warranted. Objective: To investigate whether acute ethanol intake activates the vascular RhoA/Rho kinase pathway in resistance arteries and the role of NAD(PH oxidase-derived reactive oxygen species (ROS on such response. We also evaluated the requirement of p47phox translocation for ethanol-induced NAD(PH oxidase activation. Methods: Male Wistar rats were orally treated with ethanol (1g/kg, p.o. gavage or water (control. Some rats were treated with vitamin C (250 mg/kg, p.o. gavage, 5 days before administration of water or ethanol. The mesenteric arterial bed (MAB was collected 30 min after ethanol administration. Results: Vitamin C prevented ethanol-induced increase in superoxide anion (O2- generation and lipoperoxidation in the MAB. Catalase and superoxide dismutase activities and the reduced glutathione, nitrate and hydrogen peroxide (H2O2 levels were not affected by ethanol. Vitamin C and 4-methylpyrazole prevented the increase on O2- generation induced by ethanol in cultured MAB vascular smooth muscle cells. Ethanol had no effect on phosphorylation levels of protein kinase B (Akt and eNOS (Ser1177 or Thr495 residues or MAB vascular reactivity. Vitamin C prevented ethanol-induced increase in the membrane: cytosol fraction ratio of p47phox and RhoA expression in the rat MAB. Conclusion: Acute ethanol intake induces activation of the RhoA/Rho kinase pathway by a mechanism that involves ROS generation. In resistance arteries, ethanol activates NAD(PH oxidase by inducing p47phox translocation by a redox-sensitive mechanism.

  3. Coulometric bioelectrocatalytic reactions based on NAD-dependent dehydrogenases in tricarboxylic acid cycle

    International Nuclear Information System (INIS)

    Fukuda, Jun; Tsujimura, Seiya; Kano, Kenji

    2008-01-01

    This paper describes the characterization of mediated electro-enzymatic electrolysis systems based on NAD-dependent dehydrogenase reactions in the tricarboxylic acid (TCA) cycle. A micro-bulk electrolysis system with a carbon felt anode immersed in an electrolysis solution with a value of about 10 μL was constructed for coulometric analysis of the substrate oxidation. Diaphorase (DI) was used to couple the NAD-dependent dehydrogenase reaction with the anode reaction of a suitable redox mediator. We focused on three types of NAD-dependant dehydrogenases reactions in this research: (1) isocitrate oxidation, in which the standard Gibbs energy change (ΔG o ') is negative; (2) α-ketoglutarate oxidation, which involves an electrochemically active coenzyme A (CoA); and (3) malate oxidation, which is thermodynamically unfavorable because of a large positive ΔG o ' value. The complete electrolysis of isocitrate was easily achieved, supporting the effective re-oxidation of NADH in the diaphorase-catalyzed electrochemical reaction. CoA was unfavorably oxidized at the electrodes in the presence of some mediators. The electrocatalytic oxidation of CoA was suppressed and the quantitative electrochemical oxidation of α-ketoglutarate was achieved by selecting a suitable mediator with negligibly slow electron transfer kinetics with CoA. The uphill malate oxidation was susceptible to product inhibition in the bioelectrochemical system, although NADH generated in the malate dehydrogenase reaction was immediately oxidized in the electrochemical system. The inhibition was successfully suppressed by linking citrate synthase to quench oxaloacetate and to make the total ΔG o ' value negative

  4. Status of the KM3NeT project

    International Nuclear Information System (INIS)

    Margiotta, A

    2014-01-01

    KM3NeT is a deep-sea research infrastructure being constructed in the Mediterranean Sea. It will be installed at three sites: KM3NeT-Fr, offshore Toulon, France, KM3NeT-It, offshore Portopalo di Capo Passero, Sicily (Italy) and KM3NeT-Gr, offshore Pylos, Peloponnese, Greece. It will host the next generation Cherenkov neutrino telescope and nodes for a deep sea multidisciplinary observatory, providing oceanographers, marine biologists, and geophysicists with real time measurements. The neutrino telescope will search for Galactic and extra-Galactic sources of neutrinos, complementing IceCube in its field of view. The detector will have a modular structure and consists of six building blocks, each including about one hundred Detection Units (DUs). Each DU will be equipped with 18 multi-PMT digital optical modules. The first phase of construction has started and shore and deep-sea infrastructures hosting the future KM3NeT detector are being prepared in France near Toulon and in Italy, near Capo Passero in Sicily. The technological solutions for KM3NeT and the expected performance of the detector are presented and discussed

  5. Comparison of electromagnetic and nuclear dissociation of 17Ne

    Science.gov (United States)

    Wamers, F.; Marganiec, J.; Aksouh, F.; Aksyutina, Yu.; Alvarez-Pol, H.; Aumann, T.; Beceiro-Novo, S.; Bertulani, C. A.; Boretzky, K.; Borge, M. J. G.; Chartier, M.; Chatillon, A.; Chulkov, L. V.; Cortina-Gil, D.; Emling, H.; Ershova, O.; Fraile, L. M.; Fynbo, H. O. U.; Galaviz, D.; Geissel, H.; Heil, M.; Hoffmann, D. H. H.; Hoffman, J.; Johansson, H. T.; Jonson, B.; Karagiannis, C.; Kiselev, O. A.; Kratz, J. V.; Kulessa, R.; Kurz, N.; Langer, C.; Lantz, M.; Le Bleis, T.; Lehr, C.; Lemmon, R.; Litvinov, Yu. A.; Mahata, K.; Müntz, C.; Nilsson, T.; Nociforo, C.; Ott, W.; Panin, V.; Paschalis, S.; Perea, A.; Plag, R.; Reifarth, R.; Richter, A.; Riisager, K.; Rodriguez-Tajes, C.; Rossi, D.; Savran, D.; Schrieder, G.; Simon, H.; Stroth, J.; Sümmerer, K.; Tengblad, O.; Typel, S.; Weick, H.; Wiescher, M.; Wimmer, C.

    2018-03-01

    The Borromean drip-line nucleus 17Ne has been suggested to possess a two-proton halo structure in its ground state. In the astrophysical r p -process, where the two-proton capture reaction 15O(2 p ,γ )17Ne plays an important role, the calculated reaction rate differs by several orders of magnitude between different theoretical approaches. To add to the understanding of the 17Ne structure we have studied nuclear and electromagnetic dissociation. A 500 MeV/u 17Ne beam was directed toward lead, carbon, and polyethylene targets. Oxygen isotopes in the final state were measured in coincidence with one or two protons. Different reaction branches in the dissociation of 17Ne were disentangled. The relative populations of s and d states in 16F were determined for light and heavy targets. The differential cross section for electromagnetic dissociation (EMD) shows a continuous internal energy spectrum in the three-body system 15O+2 p . The 17Ne EMD data were compared to current theoretical models. None of them, however, yields satisfactory agreement with the experimental data presented here. These new data may facilitate future development of adequate models for description of the fragmentation process.

  6. Transcriptional activation of NAD+-dependent protein deacetylase SIRT1 by nuclear receptor TLX

    International Nuclear Information System (INIS)

    Iwahara, Naotoshi; Hisahara, Shin; Hayashi, Takashi; Horio, Yoshiyuki

    2009-01-01

    An orphan nuclear receptor TLX is a transcriptional repressor that promotes the proliferation and self-renewal of neural precursor cells (NPCs). SIRT1, an NAD + -dependent protein deacetylase, is highly expressed in the NPCs and participates in neurogenesis. Here, we found that TLX colocalized with SIRT1 and knockdown of TLX by small interfering RNAs decreased SIRT1 levels in NPCs. TLX increased the SIRT1 expression by binding to the newly identified TLX-activating element in the SIRT1 gene promoter in HEK293 cells. Thus, TLX is an inducer of SIRT1 and may contribute to neurogenesis both as a transactivator and as a repressor.

  7. "Tshto zhe budjet s rodinoi s nami?" / Jevgeni Golikov

    Index Scriptorium Estoniae

    Golikov, Jevgeni

    2004-01-01

    Balti Vene Uuringute keskuse korraldatud seminaril "Venemaa aegade rägastikus: autoritarismist totalitarismini". Seminari avas Riigikogu liige Marko Mihkelson. Esinesid Vladimir Jushkin, Jevgeni Volk ja Aleksandr Sharavin

  8. Ne2 encodes protein(s) and the altered RuBisCO could be the ...

    Indian Academy of Sciences (India)

    Si Rui Pan

    2017-06-17

    Jun 17, 2017 ... Wheat hybrid necrosis is caused by the interaction of two dominant complementary genes, Ne1 and Ne2, located on ... hypothetical model presents the RuBisCO pathway of hybrid necrosis in wheat and explains how Ne1 and Ne2 interact ... were rinsed with distilled water, then cut/wrapped/labelled.

  9. Characterization of a novel thermotolerant NAD+-dependent formate dehydrogenase from hot climate plant cotton (Gossypium hirsutumL.).

    Science.gov (United States)

    Kurt-Gür, Günseli; Ordu, Emel

    2018-03-01

    NAD + -dependent formate dehydrogenases (FDH, EC 1.2.1.2), providing energy to the cell in methylotrophic microorganisms, are stress proteins in higher plants and the level of FDH expression increases under several abiotic and biotic stress conditions. They are biotechnologically important enzymes in NAD(P)H regeneration as well as CO 2 reduction. Here, the truncated form of the Gossypium hirsutum fdh1 cDNA was cloned into pQE-2 vector, and overexpressed in Escherichia coli DH5α-T1 cells. Recombinant GhFDH1 was purified 26.3-fold with a yield of 87.3%. Optimum activity was observed at pH 7.0, when substrate is formate. Kinetic analyses suggest that GhFDH1 has considerably high affinity to formate (0.76 ± 0.07 mM) and NAD + (0.06 ± 0.01 mM). At the same time, the affinity (1.98 ± 0.4 mM) and catalytic efficiency (0.0041) values of the enzyme for NADP + show that GhFDH1 is a valuable enzyme for protein engineering studies that is trying to change the coenzyme preference from NAD to NADP which has a much higher cost than that of NAD. Improving the NADP specificity is important for NADPH regeneration which is an important coenzyme used in many biotechnological production processes. The T m value of GhFDH1 is 53.3 °C and the highest enzyme activity is measured at 30 °C with a half-life of 61 h. Whilst further improvements are still required, the obtained results show that GhFDH1 is a promising enzyme for NAD(P)H regeneration for its prominent thermostability and NADP + specificity.

  10. NAD(P)H:(Quinone-Acceptor) Oxidoreductase of Tobacco Leaves Is a Flavin Mononucleotide-Containing Flavoenzyme.

    Science.gov (United States)

    Sparla, F.; Tedeschi, G.; Trost, P.

    1996-09-01

    The soluble NAD(P)H:(quinone-acceptor) oxidoreductase [NAD(P)H-QR, EC 1.6.99.2] of Nicotiana tabacum L. leaves and roots has been purified. NAD(P)H-QR contains noncovalently bound flavin mononucleotide. Pairs of subunits of 21.4 kD are linked together by disulfide bridges, but the active enzyme is a homotetramer of 94 to 100 kD showing an isoelectric point of 5.1. NAD(P)H-QR is a B-stereospecific dehydrogenase. NADH and NADPH are electron donors of similar efficiency with Kcat:Km ratios (with duroquinone) of 6.2 x 107 and 8.0 x 107 m-1 s-1, respectively. Hydrophilic quinones are good electron acceptors, although ferricyanide and dichlorophenolindophenol are also reduced. The quinones are converted to hydroquinones by an obligatory two-electron transfer. No spectral evidence for a flavin semiquinone was detected following anaerobic photoreduction. Cibacron blue and 7-iodo-acridone-4-carboxylic acid are inhibitory. Tobacco NAD(P)H-QR resembles animal DT-diaphorase in some respects (identical reaction mechanism with a two-electron transfer to quinones, unusually high catalytic capability, and donor and acceptor substrate specificity), but it differs from DT-diaphorase in molecular structure, flavin cofactor, stereospecificity, and sensitivity to inhibitors. As in the case with DT-diaphorase in animals, the main NAD(P)H-QR function in plant cells may be the reduction of quinones to quinols, which prevents the production of semiquinones and oxygen radicals. The enzyme appears to belong to a widespread group of plant and fungal flavoproteins found in different cell compartments that are able to reduce quinones.

  11. The dicotyledonous NAD malic enzyme C4 plant Cleome gynandra displays age-dependent plasticity of C4 decarboxylation biochemistry.

    Science.gov (United States)

    Sommer, M; Bräutigam, A; Weber, A P M

    2012-07-01

    The C(4) photosynthetic pathway enriches carbon dioxide in the vicinity of Rubisco, thereby enabling plants to assimilate carbon more efficiently. Three canonical subtypes of C(4) exist, named after their main decarboxylating enzymes: NAD-dependent malic enzyme type, NADP-dependent malic enzyme type and phosphoenolpyruvate carboxykinase type. Cleome gynandra is known to perform NAD-ME type C(4) photosynthesis. To further assess the mode of C(4) in C. gynandra and its manifestation in leaves of different age, total enzyme activities of eight C(4) -related enzymes and the relative abundance of 31 metabolites were measured. C. spinosa was used as a C(3) control. C. gynandra was confirmed as an NAD-ME type C(4) plant in mid-aged leaves, whereas a mixed NAD-ME and PEPCK type was observed in older leaves. Young leaves showed a C(3) -C(4) intermediate state with respect to enzyme activities and metabolite abundances. Comparative transcriptome analysis of mid-aged leaves of C. gynandra and C. spinosa showed that the transcript of only one aspartate aminotransferase (AspAT) isoform is highly abundant in C. gynandra. However, the canonical model of the NAD-ME pathway requires two AspATs, a mitochondrial and a cytosolic isoform. Surprisingly, our results indicate the existence of only one highly abundant AspAT isoform. Using GFP-fusion, this isozyme was localised exclusively to mitochondria. We propose a revised model of NAD-ME type C(4) photosynthesis in C. gynandra, in which both AspAT catalysed reactions take place in mitochondria and PEPCK catalyses an alternative decarboxylating pathway. © 2012 German Botanical Society and The Royal Botanical Society of the Netherlands.

  12. Letter of Intent to Build a MiniBooNE Near Detector: BooNE

    Energy Technology Data Exchange (ETDEWEB)

    Stancu, I. [Univ. of Alabama, Tuscaloosa, AL (United States); Djurcic, Z. [Argonne National Lab. (ANL), Argonne, IL (United States); Smith, D. [Embry-Riddle Aeronautical Univ., Prescott, AZ (United States); Ford, R. [Fermi National Accelerator Lab. (FNAL), Batavia, IL (United States); Kobilarcik, T. [Fermi National Accelerator Lab. (FNAL), Batavia, IL (United States); Marsh, W. [Fermi National Accelerator Lab. (FNAL), Batavia, IL (United States); Moore, C. D. [Fermi National Accelerator Lab. (FNAL), Batavia, IL (United States); Grange, J. [Univ. of Florida, Gainesville, FL (United States); Osmanov, B. [Univ. of Florida, Gainesville, FL (United States); Ray, H. [Univ. of Florida, Gainesville, FL (United States); Garvey, G. T. [Los Alamos National Lab. (LANL), Los Alamos, NM (United States). et al.

    2009-10-12

    There is accumulating evidence for a difference between neutrino and antineutrino oscillations at the {approx}1 eV{sup 2} scale. The MiniBooNE experiment observes an unexplained excess of electron-like events at low energies in neutrino mode, which may be due, for example, to either a neutral current radiative interaction, sterile neutrino decay, or to neutrino oscillations involving sterile neutrinos and which may be related to the LSND signal. No excess of electron-like events (-0.5 {+-} 7.8 {+-} 8.7), however, is observed so far at low energies in antineutrino mode. Furthermore, global 3+1 and 3+2 sterile neutrino fits to the world neutrino and antineutrino data suggest a difference between neutrinos and antineutrinos with significant (sin{sup 2} 2{theta}{sub {mu}{mu}} {approx} 35%) {bar {nu}}{sub {mu}} disappearance. In order to test whether the low-energy excess is due to neutrino oscillations and whether there is a difference between {nu}{sub {mu}} and {bar {nu}}{sub {mu}} disappearance, we propose building a second MiniBooNE detector at (or moving the existing MiniBooNE detector to) a distance of {approx}200 m from the Booster Neutrino Beam (BNB) production target. With identical detectors at different distances, most of the systematic errors will cancel when taking a ratio of events in the two detectors, as the neutrino flux varies as 1/r{sup 2} to a calculable approximation. This will allow sensitive tests of oscillations for both {nu}{sub e} and {bar {nu}} appearance and {nu}{sub {mu}} and {bar {nu}}{sub {mu}} disappearance. Furthermore, a comparison between oscillations in neutrino mode and antineutrino mode will allow a sensitive search for CP and CPT violation in the lepton sector at short baseline ({Delta}m{sup 2} > 0.1 eV{sup 2}). Finally, by comparing the rates for a neutral current (NC) reaction, such as NC {pi}{sup 0} scattering or NC elastic scattering, a direct search for sterile neutrinos will be made. The initial amount of running time

  13. ATP- and NAD+-dependent DNA ligases share an essential function in the halophilic archaeon Haloferax volcanii

    DEFF Research Database (Denmark)

    Zhao, A.; Gray, F. C; MacNeill, S. A.

    2006-01-01

    DNA ligases join the ends of DNA molecules during replication, repair and recombination. ATP-dependent ligases are found predominantly in the eukarya and archaea whereas NAD+-dependent DNA ligases are found only in the eubacteria and in entomopoxviruses. Using the genetically tractable halophile....... volcanii also encodes an NAD+-dependent DNA ligase family member, LigN, the first such enzyme to be identified in the archaea, and present phylogenetic analysis indicating that the gene encoding this protein has been acquired by lateral gene transfer (LGT) from eubacteria. As with LigA, we show that Lig...

  14. Rev1 contributes to proper mitochondrial function via the PARP-NAD(+)-SIRT1-PGC1 alpha axis

    DEFF Research Database (Denmark)

    Fakouri, Nima Borhan; Durhuus, Jon Ambaek; Regnell, Christine Elisabeth

    2017-01-01

    (ADP) ribose polymerase 1 (PARP1) activity, low endogenous NAD+, low expression of SIRT1 and PGC1α and low adenosine monophosphate (AMP)-activated kinase (AMPK) activity. We conclude that replication stress via Rev1-deficiency contributes to metabolic stress caused by compromized mitochondrial function via...... the PARP-NAD+-SIRT1-PGC1α axis.......- and double-stranded DNA breaks (SSBs and DSBs), and single-stranded gaps can block progression of the DNA replication fork, causing replicative stress and/or cell cycle arrest. However, translesion synthesis (TLS) DNA polymerases, such as Rev1, have the ability to bypass some DNA lesions, which can...

  15. Characterization of NAD salvage pathways and their role in virulence in Streptococcus pneumoniae

    Science.gov (United States)

    Johnson, Michael D. L.; Echlin, Haley; Dao, Tina H.

    2015-01-01

    NAD is a necessary cofactor present in all living cells. Some bacteria cannot de novo synthesize NAD and must use the salvage pathway to import niacin or nicotinamide riboside via substrate importers NiaX and PnuC, respectively. Although homologues of these two importers and their substrates have been identified in other organisms, limited data exist in Streptococcus pneumoniae, specifically, on its effect on overall virulence. Here, we sought to characterize the substrate specificity of NiaX and PnuC in Str. pneumoniae TIGR4 and the contribution of these proteins to virulence of the pathogen. Although binding affinity of each importer for nicotinamide mononucleotide may overlap, we found NiaX to specifically import nicotinamide and nicotinic acid, and PnuC to be primarily responsible for nicotinamide riboside import. Furthermore, a pnuC mutant is completely attenuated during both intranasal and intratracheal infections in mice. Taken together, these findings underscore the importance of substrate salvage in pneumococcal pathogenesis and indicate that PnuC could potentially be a viable small-molecule therapeutic target to alleviate disease progression in the host. PMID:26311256

  16. Purification, kinetic behavior, and regulation of NAD(P)+ malic enzyme of tumor mitochondria.

    Science.gov (United States)

    Moreadith, R W; Lehninger, A L

    1984-05-25

    The purification and kinetic characterization of an NAD(P)+-malic enzyme from 22aH mouse hepatoma mitochondria are described. The enzyme was purified 328-fold with a final yield of 51% and specific activity of 38.1 units/mg of protein by employing DEAE-cellulose chromatography and an ATP affinity column. Sephadex G-200 chromatography yielded a native Mr = 240,000. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed a major subunit with Mr = 61,000, suggesting a tetrameric structure, and also showed that the preparation contained less than 10% polypeptide impurities. Use of the ATP affinity column required the presence of MnCl2 and fumarate (an allosteric activator) in the elution buffers. In the absence of fumarate, the Michaelis constants for malate, NAD+, and NADP+ were 3.6 mM, 55 microM, and 72 microM, respectively; in the presence of fumarate (2 mM), the constants were 0.34 mM, 9 microM, and 13 microM, respectively. ATP was shown to be an allosteric inhibitor, competitive with malate. However, the inhibition by ATP displayed hyperbolic competitive kinetics with a KI (ATP) of 80 microM (minus fumarate) and 0.5 mM (plus 2 mM fumarate). The allosteric properties of the enzyme are integrated into a rationale for its specific role in the pathways of malate and glutamate oxidation in tumor mitochondria.

  17. Excavations of an Early Neolithic Site at Tăşnad, Romania

    Directory of Open Access Journals (Sweden)

    Ciprian Astaloș

    2013-10-01

    Full Text Available The town of Tăşnad, in north-west Romania, is situated at the western end of the Tăşnad Hills which rise to a height of up to 230m above sea-level; the site ‘Sere’ is situated south-west of the town near a thermal spa on the banks of the Cehal river, a tributary of the Ier. The Cehal valley opens towards the Ier and Someş plains which form the north-easternmost part of the Great Hungarian Plain, a marshy area until the large-scale drainage-works of the 19th and 20th centuries. Even today, the Cehal valley is quite swampy, especially at the confluence with the Ier. The Austrian military maps demonstrate large-scale forest-clearance during the last three centuries; at the end of the 18th century, the site itself was still forested. Several prehistoric sites from different periods are located on the first and second terraces of the Cehal, at altitudes of around 140m.

  18. Declining NAD+ Induces a Pseudohypoxic State Disrupting Nuclear-Mitochondrial Communication during Aging

    Science.gov (United States)

    Gomes, Ana P.; Price, Nathan L.; Ling, Alvin J.Y.; Moslehi, Javid J.; Montgomery, Magdalene K.; Rajman, Luis; White, James P.; Teodoro, João S.; Wrann, Christiane D.; Hubbard, Basil P.; Mercken, Evi M.; Palmeira, Carlos M.; de Cabo, Rafael; Rolo, Anabela P.; Turner, Nigel; Bell, Eric L.; Sinclair, David A.

    2014-01-01

    SUMMARY Ever since eukaryotes subsumed the bacterial ancestor of mitochondria, the nuclear and mitochondrial genomes have had to closely coordinate their activities, as each encode different subunits of the oxidative phosphorylation (OXPHOS) system. Mitochondrial dysfunction is a hallmark of aging, but its causes are debated. We show that, during aging, there is a specific loss of mitochondrial, but not nuclear, encoded OXPHOS subunits. We trace the cause to an alternate PGC-1α/β-independent pathway of nuclear-mitochondrial communication that is induced by a decline in nuclear NAD+ and the accumulation of HIF-1α under normoxic conditions, with parallels to Warburg reprogramming. Deleting SIRT1 accelerates this process, whereas raising NAD+ levels in old mice restores mitochondrial function to that of a young mouse in a SIRT1-dependent manner. Thus, a pseudohypoxic state that disrupts PGC-1α/β-independent nuclear-mitochondrial communication contributes to the decline in mitochondrial function with age, a process that is apparently reversible. PMID:24360282

  19. Declining NAD(+) induces a pseudohypoxic state disrupting nuclear-mitochondrial communication during aging.

    Science.gov (United States)

    Gomes, Ana P; Price, Nathan L; Ling, Alvin J Y; Moslehi, Javid J; Montgomery, Magdalene K; Rajman, Luis; White, James P; Teodoro, João S; Wrann, Christiane D; Hubbard, Basil P; Mercken, Evi M; Palmeira, Carlos M; de Cabo, Rafael; Rolo, Anabela P; Turner, Nigel; Bell, Eric L; Sinclair, David A

    2013-12-19

    Ever since eukaryotes subsumed the bacterial ancestor of mitochondria, the nuclear and mitochondrial genomes have had to closely coordinate their activities, as each encode different subunits of the oxidative phosphorylation (OXPHOS) system. Mitochondrial dysfunction is a hallmark of aging, but its causes are debated. We show that, during aging, there is a specific loss of mitochondrial, but not nuclear, encoded OXPHOS subunits. We trace the cause to an alternate PGC-1α/β-independent pathway of nuclear-mitochondrial communication that is induced by a decline in nuclear NAD(+) and the accumulation of HIF-1α under normoxic conditions, with parallels to Warburg reprogramming. Deleting SIRT1 accelerates this process, whereas raising NAD(+) levels in old mice restores mitochondrial function to that of a young mouse in a SIRT1-dependent manner. Thus, a pseudohypoxic state that disrupts PGC-1α/β-independent nuclear-mitochondrial communication contributes to the decline in mitochondrial function with age, a process that is apparently reversible. Copyright © 2013 Elsevier Inc. All rights reserved.

  20. Discoveries of nicotinamide riboside as a nutrient and conserved NRK genes establish a Preiss-Handler independent route to NAD+ in fungi and humans.

    Science.gov (United States)

    Bieganowski, Pawel; Brenner, Charles

    2004-05-14

    NAD+ is essential for life in all organisms, both as a coenzyme for oxidoreductases and as a source of ADPribosyl groups used in various reactions, including those that retard aging in experimental systems. Nicotinic acid and nicotinamide were defined as the vitamin precursors of NAD+ in Elvehjem's classic discoveries of the 1930s. The accepted view of eukaryotic NAD+ biosynthesis, that all anabolism flows through nicotinic acid mononucleotide, was challenged experimentally and revealed that nicotinamide riboside is an unanticipated NAD+ precursor in yeast. Nicotinamide riboside kinases from yeast and humans essential for this pathway were identified and found to be highly specific for phosphorylation of nicotinamide riboside and the cancer drug tiazofurin. Nicotinamide riboside was discovered as a nutrient in milk, suggesting that nicotinamide riboside is a useful compound for elevation of NAD+ levels in humans.

  1. Shape evolution of Ne isotopes and Ne hypernuclei: The interplay of pairing and tensor interactions

    Directory of Open Access Journals (Sweden)

    Li A.

    2014-03-01

    Full Text Available We study tensor and pairing effects on the quadruple deformation of neon isotopes based on a deformed Skyrme-Hartree-Fock model with BCS approximation for the pairing channel. We extend the Skyrme-Hartree-Fock formalism for the description of hypernuclei adopting the recently-proposed ESC08b hyperon-nucleon interaction. It is found that the interplay of pairing and tensor interactions is crucial to derive the deformations in several neon isotopes. Especially, the shapes of 26,30Ne are studied in details in comparisons with experimentally observed shapes. Furthermore the deformations of the hypernuclei are compared with the corresponding neon isotopic cores in the presence of tensor force. We find the same shapes with somewhat smaller deformations for single Λ-hypernuclei compared with their core deformations.

  2. Flaking and blistering on He and Ne bombardments

    International Nuclear Information System (INIS)

    Kamada, K.; Naramoto, H.

    1979-01-01

    Large scale exfoliation formed by 300 keV He + bombardment of niobium without any preceding blistering is investigated, in comparison with the blistering due to 450 and 850 keV Ne + bombardments. In-situ observations of the erosion processes were performed in a scanning electron microscope connected to the Van de Graaff. Critical doses of 7.2 x 10 17 He + /cm 2 , 2.4 x 10 17 Ne + /cm 2 and 4.0 x 10 17 Ne + /cm 2 were obtained for the 300 keV He flaking, 450 keV Ne blistering and 850 keV Ne blistering, respectively. The He flaking was presumed to be due to brittle fashion peeling-off of the surface layer by the bending moment driven by the internal gas pressure. The blistering, on the other hand, was presumed to be the result of the ductile fashion spreading of the lenticular bubble in the sub-surface layer. The necessary pressure for the peeling-off of the cover was calculated, and was speculated to be able to work as the driving force for the flaking from its unexpectedly low values. Fractographies under the exfoliations were discussed for both flaking and blistering. (author)

  3. The KM3NeT project: status and perspectives

    Directory of Open Access Journals (Sweden)

    A. Margiotta

    2013-01-01

    Full Text Available KM3NeT is an international consortium involving more than 300 scientists from 10 EU countries. Its main objective is the construction of a multi-km3 high-energy neutrino telescope in the Mediterranean Sea that will also host an interdisciplinary observatory for marine sciences. KM3NeT has been included in the roadmap of the European Strategy Forum of Research Infrastructures (ESFRI. Very high energy neutrinos are important messengers to study non-thermal phenomena in the Universe. The pioneering ANTARES, NEMO and NESTOR underwater neutrino telescope projects include the extensive R&D knowledge base behind the KM3NeT project. A Technical Design Report has been published that describes the technological solutions chosen for the detector. The present status of the project is presented.

  4. The Fermilab Short-Baseline Program: MicroBooNE

    Energy Technology Data Exchange (ETDEWEB)

    Schukraft, Anne [Fermi National Accelerator Lab. (FNAL), Batavia, IL (United States)

    2016-01-01

    The MicroBooNE experiment is the first of three detectors of the Fermilab short-baseline neutrino program that started operation in the Booster Neutrino Beamline in October 2015 [1]. When completed, the three-detector lineup will explore short-baseline neutrino oscillations and will be sensitive to sterile neutrino scenarios. MicroBooNE in itself is now starting its own physics program, with the measurement of neutrino-argon cross sections in the ~1GeV range being one of its main physics goals. These proceedings describe the status of the detector, the start of operation, and the automated reconstruction of the first neutrino events observed with MicroBooNE. Prospects for upcoming cross section measurements are also given.

  5. New low pressure (LP) turbines for NE Krsko

    International Nuclear Information System (INIS)

    Nemcic, K.; Novsak, M.

    2004-01-01

    During the evaluation of possible future maintenance strategies on steam turbine in very short period of time, engineering decision was made by NE Krsko in agreement with Owners to replace the existing two Low Pressure (LP) Turbines with new upgrading LP Turbines. This decision is presented with review of the various steam turbine problems as: SCC on turbine discs; blades cracking; erosion-corrosion with comparison of various maintenance options and efforts undertaken by the NE Krsko to improve performance of the original low pressure turbines. This paper presents the NEK approach to solve the possible future problems with steam turbine operation in NE Krsko as pro-active engineering and maintenance activities on the steam turbine. This paper also presents improvements involving retrofits, confined to the main steam turbine path, with major differences between original and new LP Turbines as beneficial replacement because of turbine MWe upgrading and return capital expenditures.(author)

  6. Bystřice nad Perštejnem a útlum těžby uranu

    Czech Academy of Sciences Publication Activity Database

    Vaishar, Antonín; Kallabová, Eva; Zapletalová, Jana

    2002-01-01

    Roč. 5, č. 3 (2002), s. 8-16 ISSN 1212-0855 R&D Projects: GA AV ČR KSK3046108 Keywords : Bystřice nad Perštejnem * uranium mining * industrial restructing Subject RIV: DE - Earth Magnetism, Geodesy, Geography

  7. Restoration of Mitochondrial NAD+Levels Delays Stem Cell Senescence and Facilitates Reprogramming of Aged Somatic Cells.

    Science.gov (United States)

    Son, Myung Jin; Kwon, Youjeong; Son, Taekwon; Cho, Yee Sook

    2016-12-01

    The fundamental tenet that aging is irreversible has been challenged by the development of reprogramming technology that can restore molecular and cellular age by reversing the progression of aging. The use of cells from aged individuals as sources for reprogramming or transplantation creates a major barrier in stem cell therapy with respect to cell quality and quantity. Here, we investigated the molecular features underlying senescence and rejuvenation during aged cell reprogramming and identified novel factors that can overcome age-associated barriers. Enzymes, such as nicotinamide nucleotide transhydrogenase (NNT) and nicotinamide mononucleotide adenylyltransferase 3 (NMNAT3), that control mitochondrial NAD + levels appear to be susceptible to aging. In aged cells, mitochondrial NAD + levels decrease, accompanied by reduced SIRT3 activity; these changes severely impede cell fate transition. However, in cells collected from aged p16 knockout mice, which exhibit delayed cellular senescence, no changes in NNT or NMNAT3 expression were found. Importantly, restoring mitochondrial NAD + levels by overexpressing NNT and NMNAT3 enhanced reprogramming efficiency of aged somatic cells and extended the lifespan of human mesenchymal stem cells by delaying replicative senescence. These results demonstrate that maintenance of mitochondrial NAD + levels is critical for reversing the mechanisms of aging and ensuring that cells collected from aged individuals are of high quality. Stem Cells 2016;34:2840-2851. © 2016 AlphaMed Press.

  8. Archeologický výzkum hradu Kamýk nad Vltavou v roce 2009

    Czech Academy of Sciences Publication Activity Database

    Durdík, Tomáš

    2010-01-01

    Roč. 118, č. 1 (2010), s. 1-16 ISSN 1803-1382 R&D Projects: GA MK DB06P01OPP004 Institutional research plan: CEZ:AV0Z80020508 Keywords : castle * castellology * Kamýk nad Vltavou * medieval archaeology * architecture * Middle Ages * Bohemia Subject RIV: AC - Archeology, Anthropology, Ethnology

  9. Phenotypic and genetic characterization of NAD-dependent Pasteurellaceae from the respiratory tract of pigs and their possible pathogenetic importance

    DEFF Research Database (Denmark)

    Kielstein, P.; Wuthe, H.H.; Angen, Øystein

    2001-01-01

    . In the present study, 107 of these NAD-dependent isolates from the porcine respiratory tract, primarily from lungs with pathological changes, were investigated. On the basis of phenotypic criteria, such as haemolysis, urease, catalase, and indole formation as well as other fermentative activities, 50...

  10. Kuidas saab Tallinna Vee brittidelt tagasi osta, kui nad ei taha müüa? / Andres Reimer

    Index Scriptorium Estoniae

    Reimer, Andres

    2009-01-01

    Autori sõnul peavad tõsiselt võetavad analüütikud ja arvamusliidrid Tallinna Vee aktsiate linnale ostmist majanduslikult põhjendamata sammuks, mis tooks kaasa suure laenukoorma, mille lõpuks maksavad kinni tallinlased. Asjatundjate hinnangul ei paista Tallinna Vee omanikel olevat ühtegi põhjust, miks nad tahaksid firma aktsiaid müüa

  11. Millest nad kirjutavad ehk Näidendivõistluse tänavusügisene saak / Triin Sinissaar

    Index Scriptorium Estoniae

    Sinissaar, Triin

    2003-01-01

    I preemia: Urmas Lennuk, "Boob teab"ja Jaan Undusk, "Quevedo"; II preemia: Anu Allas, "Lendav rõdu ehk Nagu nad tahtsid" ja Raivo Kütt, "Papa" ("Sõtse ja venna"); III preemia Jakob Karu, "Asjade seis" ja Hans Nordberg, "aaron : juuni"; ergutuspreemia: Urmas Lennuk, "Kadunud kindapood", Urmas Vadi, "Kadunud kosmoses" ja Jaan Võõramaa, "Mamma"

  12. The differences between NAD-ME and NADP-ME subtypes of C4 photosynthesis: more than decarboxylating enzymes

    Directory of Open Access Journals (Sweden)

    Xiaolan Rao

    2016-10-01

    Full Text Available As an adaptation to changing climatic conditions that caused high rates of photorespiration, C4 plants have evolved to display higher photosynthetic efficiency than C3 plants under elevated temperature, high light intensities and drought. The C4 plants independently evolved more than 60 times in 19 families of angiosperms to establish similar but not uniform C4 mechanisms to concentrate CO2 around the carboxylating enzyme Rubisco. C4 photosynthesis is divided into at least two basic biochemical subtypes based on the primary decarboxylating enzymes, NAD-dependent malic enzyme (NAD-ME and NADP-dependent malic enzyme (NADP-ME. The multiple polygenetic origins of these subtypes raise questions about the association of C4 variation between biochemical subtypes and diverse lineages. This review addresses the differences in evolutionary scenario, leaf anatomy, and especially C4 metabolic flow, C4 transporters and cell-specific function deduced from recently reported cell-specific transcriptomic, proteomic and metabolic analyses of NAD-ME and NADP-ME subtypes. Current omic analysis has revealed the extent to which component abundances differ between the two biochemical subtypes, leading to a better understanding of C4 photosynthetic mechanisms in NAD-ME and NADP-ME subtypes.

  13. Environment Dictates Dependence on Mitochondrial Complex I for NAD+ and Aspartate Production and Determines Cancer Cell Sensitivity to Metformin.

    Science.gov (United States)

    Gui, Dan Y; Sullivan, Lucas B; Luengo, Alba; Hosios, Aaron M; Bush, Lauren N; Gitego, Nadege; Davidson, Shawn M; Freinkman, Elizaveta; Thomas, Craig J; Vander Heiden, Matthew G

    2016-11-08

    Metformin use is associated with reduced cancer mortality, but how metformin impacts cancer outcomes is controversial. Although metformin can act on cells autonomously to inhibit tumor growth, the doses of metformin that inhibit proliferation in tissue culture are much higher than what has been described in vivo. Here, we show that the environment drastically alters sensitivity to metformin and other complex I inhibitors. We find that complex I supports proliferation by regenerating nicotinamide adenine dinucleotide (NAD)+, and metformin's anti-proliferative effect is due to loss of NAD+/NADH homeostasis and inhibition of aspartate biosynthesis. However, complex I is only one of many inputs that determines the cellular NAD+/NADH ratio, and dependency on complex I is dictated by the activity of other pathways that affect NAD+ regeneration and aspartate levels. This suggests that cancer drug sensitivity and resistance are not intrinsic properties of cancer cells, and demonstrates that the environment can dictate sensitivity to therapies that impact cell metabolism. Copyright © 2016 Elsevier Inc. All rights reserved.

  14. Pharmacological inhibition of nicotinamide phosphoribosyltransferase/visfatin enzymatic activity identifies a new inflammatory pathway linked to NAD.

    Directory of Open Access Journals (Sweden)

    Nathalie Busso

    Full Text Available Nicotinamide phosphoribosyltransferase (NAMPT, also known as visfatin, is the rate-limiting enzyme in the salvage pathway of NAD biosynthesis from nicotinamide. Since its expression is upregulated during inflammation, NAMPT represents a novel clinical biomarker in acute lung injury, rheumatoid arthritis, and Crohn's disease. However, its role in disease progression remains unknown. We report here that NAMPT is a key player in inflammatory arthritis. Increased expression of NAMPT was confirmed in mice with collagen-induced arthritis, both in serum and in the arthritic paw. Importantly, a specific competitive inhibitor of NAMPT effectively reduced arthritis severity with comparable activity to etanercept, and decreased pro-inflammatory cytokine secretion in affected joints. Moreover, NAMPT inhibition reduced intracellular NAD concentration in inflammatory cells and circulating TNFalpha levels during endotoxemia in mice. In vitro pharmacological inhibition of NAMPT reduced the intracellular concentration of NAD and pro-inflammatory cytokine secretion by inflammatory cells. Thus, NAMPT links NAD metabolism to inflammatory cytokine secretion by leukocytes, and its inhibition might therefore have therapeutic efficacy in immune-mediated inflammatory disorders.

  15. Decarboxylation of Malate in the Crassulacean Acid Metabolism Plant Bryophyllum (Kalanchoe) fedtschenkoi (Role of NAD-Malic Enzyme).

    Science.gov (United States)

    Cook, R. M.; Lindsay, J. G.; Wilkins, M. B.; Nimmo, H. G.

    1995-01-01

    The role of NAD-malic enzyme (NAD-ME) in the Crassulacean acid metabolism plant Bryophyllum (Kalanchoe) fedtschenkoi was investigated using preparations of intact and solubilized mitochondria from fully expanded leaves. Intact, coupled mitochondria isolated during the day or night did not differ in their ability to take up [14C]malic acid from the surrounding medium or to respire using malate or succinate as substrate. However, intact mitochondria isolated from plants during the day decarboxylated added malate to pyruvate significantly faster than mitochondria isolated from plants at night. NAD-ME activity in solubilized mitochondrial extracts showed hysteretic kinetics and was stimulated by a number of activators, including acetyl-coenzyme A, fructose-1,6-bisphosphate, and sulfate ions. In the absence of these effectors, reaction progress curves were nonlinear, with a pronounced acceleration phase. The lag period before a steady-state rate was reached in assays of mitochondrial extracts decreased during the photoperiod and increased slowly during the period of darkness. However, these changes in the kinetic properties of the enzyme could not account for the changes in the rate of decarboxylation of malate by intact mitochondria. Gel-filtration experiments showed that mitochondrial extracts contained three forms of NAD-ME with different molecular weights. The relative proportions of the three forms varied somewhat throughout the light/dark cycle, but this did not account for the changes in the kinetics behavior of the enzyme during the diurnal cycle. PMID:12228671

  16. Level-resolved R-matrix calculations for the electron-impact excitation of Ne3+ and Ne6+

    International Nuclear Information System (INIS)

    Ludlow, J. A.; Lee, T. G.; Ballance, C. P.; Loch, S. D.; Pindzola, M. S.

    2011-01-01

    Large-scale R-matrix calculations are carried out for the electron-impact excitation of Ne 3+ and Ne 6+ . For Ne 3+ , a 581-LSJ-level R-matrix intermediate coupling frame transformation calculation is made for excitations up to the n=4 shell. For some transitions, large effective collision strength differences are found with current 23-jKJ-level Breit-Pauli R-matrix and earlier 22-LSJ-level R-matrix jj omega (JAJOM) calculations. For Ne 6+ , a 171-jKJ-level Breit-Pauli R-matrix calculation is made for excitations up to the n=5 shell. For some transitions, large effective collision strength differences are found with current 46-jKJ-level Breit-Pauli R-matrix and earlier 46-LSJ-level R-matrix JAJOM calculations. Together with existing R-matrix calculations for other ion stages, high-quality excitation data are now available for astrophysical and laboratory plasma modeling along the entire Ne isonuclear sequence.

  17. DAΦNE Control System status and performance

    International Nuclear Information System (INIS)

    Di Pirro, G.; Drago, A.; Mazzitelli, G.; Milardi, C.; Sannibale, F.; Stecchi, A.; Stella, A.

    1998-01-01

    The DAΦNE Control System allowed the step by step commissioning of the major subsystems as they were installed, proving to be modular and extensible. Recently the guidelines of the Control System evolution concerned the development of machine operational procedures and the integration of diagnostic tools. Particular attention has been reserved to the problem of saving and restoring element data sts as well as to the DAΦNE general data handling. A system overview including installation status, features, and operation results is presented

  18. Evaluation of triggering schemes for KM3NeT

    Energy Technology Data Exchange (ETDEWEB)

    Seitz, T., E-mail: Thomas.Seitz@physik.uni-erlangen.de [Erlangen Centre for Astroparticle Physics, Erwin-Rommel-Str. 1, 91058 Erlangen (Germany); Herold, B., E-mail: Bjoern.Herold@physik.uni-erlangen.de [Erlangen Centre for Astroparticle Physics, Erwin-Rommel-Str. 1, 91058 Erlangen (Germany); Shanidze, R., E-mail: shanidze@physik.uni-erlangen.de [Erlangen Centre for Astroparticle Physics, Erwin-Rommel-Str. 1, 91058 Erlangen (Germany)

    2013-10-11

    The future neutrino telescope KM3NeT, to be built in the Mediterranean Sea, will be the largest of its kind. It will include nearly two hundred thousand photomultiplier tubes (PMT) mounted in multi-PMT digital optical modules (DOM). The dominant source of the PMT signals is decays of {sup 40}K and marine fauna bioluminescence. Selection of neutrino and muon events from this continuous optical background signals requires the implementation of fast and efficient triggers. Various schemes for the filtering of background data and the selection of neutrino and muon events were evaluated for the KM3NeT telescope using Monte Carlo simulations.

  19. Evaluation of triggering schemes for KM3NeT

    Science.gov (United States)

    Seitz, T.; Herold, B.; Shanidze, R.

    2013-10-01

    The future neutrino telescope KM3NeT, to be built in the Mediterranean Sea, will be the largest of its kind. It will include nearly two hundred thousand photomultiplier tubes (PMT) mounted in multi-PMT digital optical modules (DOM). The dominant source of the PMT signals is decays of 40K and marine fauna bioluminescence. Selection of neutrino and muon events from this continuous optical background signals requires the implementation of fast and efficient triggers. Various schemes for the filtering of background data and the selection of neutrino and muon events were evaluated for the KM3NeT telescope using Monte Carlo simulations.

  20. Cascade sensitivity studies for KM3NeT

    Directory of Open Access Journals (Sweden)

    Fusco Luigi Antonio

    2016-01-01

    Full Text Available KM3NeT is a future research infrastructure in the deep seas of the Mediterranean housing a large scale neutrino telescope. The first phase of construction of the telescope has started. Next step is an intermediate phase realising a detector volume of about one-third of the final detector volume. We report on calculations of the sensitivity of the KM3NeT detector to showering neutrino events, the strategy to optimise the detector to a cosmic neutrino flux analogous to the one reported by the IceCube Collaboration and the results of this strategy applied to the intermediate phase detector.

  1. Near-threshold electron impact ionization of Ne and Xe

    International Nuclear Information System (INIS)

    Yates, B R; Khakoo, M A; Keane, K

    2009-01-01

    Doubly differential cross-sections for the single electron impact ionization of Ne and Xe have been measured at several energies below the second ionization energy. The results indicate that the ionization of Ne is strongly influenced by the polarization of the ionized 2 2 P 3/2,1/2 core, where as this influence is significantly reduced for Xe. Single differential cross-sections are derived from the doubly differential cross-sections and for Xenon these show profiles similar to Helium ('smile'), whereas for Neon they show a dissimilar profile ('frown').

  2. Experimental ion mobility measurements in Ne-N2

    International Nuclear Information System (INIS)

    Cortez, A.F.V.; Encarnação, P.M.C.C.; Santos, F.P.; Borges, F.I.G.M.; Conde, C.A.N.; Veenhof, R.; Neves, P.N.B.

    2016-01-01

    Data on ion mobility is important to improve the performance of large volume gaseous detectors, such as the ALICE TPC or in the NEXT experiment. In the present work the method, experimental setup and results for the ion mobility measurements in Ne-N 2 mixtures are presented. The results for this mixture show the presence of two peaks for different gas ratios of Ne-N 2 , low reduced electric fields, E / N , 10–20 Td (2.4–4.8 kV·cm −1 ·bar −1 ), low pressures 6–8 Torr (8–10.6 mbar) and at room temperature.

  3. Plastic scintillator response to relativistic Ne, Ar, Fe IONS

    Science.gov (United States)

    Salamon, M. H.; Ahlen, S. P.

    1982-04-01

    The response to relativistic (0-600 MeV/amu) Ne, Ar, and Fe ions and to cosmic ray muons of four widely used commercial plastic scintillators, NE110, Pilot Y, Pilot F, and Pilot B, is discussed. Fitted expressions for scintillation efficiency for each scintillator and charge are given, and these are compared with the predictions of both the Voltz model and a modification of the Birks model. Resolution measurements demonstrate the relative roles of the halo and quenched core in heavy ion response, and point to a novel use for plastic scintillators.

  4. OD ANTROPOCENTRIČNE K EKOCENTRIČNOJ ETICI

    OpenAIRE

    Kirn, Andrej

    1992-01-01

    Sve veći ekološki problemi zahtijevaju širenje etičke regulacije čovjekova odnosa spram prirode. Tome se suprotstavlja cijela antropocentrično zasnovana europska kultura, a osobito njezina antropocentrično ograničena, agresivna ekonomija i etika. Europski etički antropocentrizam ima dva izvora: grčku kulturu i židovsko-kršćansku misao. U okviru antropocentrične etike moralnost se ograničava na ljudsku zajednicu, na zajednicu slobodnih i razumnih ljudi. Bitna ograničenost antropocentrične etik...

  5. Catalytic Properties of the Isolated Diaphorase Fragment of the NAD+-Reducing [NiFe]-Hydrogenase from Ralstonia eutropha

    Science.gov (United States)

    Lauterbach, Lars; Idris, Zulkifli; Vincent, Kylie A.; Lenz, Oliver

    2011-01-01

    The NAD+-reducing soluble hydrogenase (SH) from Ralstonia eutropha H16 catalyzes the H2-driven reduction of NAD+, as well as reverse electron transfer from NADH to H+, in the presence of O2. It comprises six subunits, HoxHYFUI2, and incorporates a [NiFe] H+/H2 cycling catalytic centre, two non-covalently bound flavin mononucleotide (FMN) groups and an iron-sulfur cluster relay for electron transfer. This study provides the first characterization of the diaphorase sub-complex made up of HoxF and HoxU. Sequence comparisons with the closely related peripheral subunits of Complex I in combination with UV/Vis spectroscopy and the quantification of the metal and FMN content revealed that HoxFU accommodates a [2Fe2S] cluster, FMN and a series of [4Fe4S] clusters. Protein film electrochemistry (PFE) experiments show clear electrocatalytic activity for both NAD+ reduction and NADH oxidation with minimal overpotential relative to the potential of the NAD+/NADH couple. Michaelis-Menten constants of 56 µM and 197 µM were determined for NADH and NAD+, respectively. Catalysis in both directions is product inhibited with K I values of around 0.2 mM. In PFE experiments, the electrocatalytic current was unaffected by O2, however in aerobic solution assays, a moderate superoxide production rate of 54 nmol per mg of protein was observed, meaning that the formation of reactive oxygen species (ROS) observed for the native SH can be attributed mainly to HoxFU. The results are discussed in terms of their implications for aerobic functioning of the SH and possible control mechanism for the direction of catalysis. PMID:22016788

  6. Crystal Structure of Human Dihydrolipoamide Dehydrogenase: NAD[superscript +]/NADH Binding and the Structural Basis of Disease-causing Mutations

    Energy Technology Data Exchange (ETDEWEB)

    Brautigam, Chad A.; Chuang, Jacinta L.; Tomchick, Diana R.; Machius, Mischa; Chuang, David T. (U. of Texas-SMED)

    2010-07-13

    Human dihydrolipoamide dehydrogenase (hE3) is an enzymatic component common to the mitochondrial {alpha}-ketoacid dehydrogenase and glycine decarboxylase complexes. Mutations to this homodimeric flavoprotein cause the often-fatal human disease known as E3 deficiency. To catalyze the oxidation of dihydrolipoamide, hE3 uses two molecules: noncovalently bound FAD and a transiently bound substrate, NAD{sup +}. To address the catalytic mechanism of hE3 and the structural basis for E3 deficiency, the crystal structures of hE3 in the presence of NAD{sup +} or NADH have been determined at resolutions of 2.5 {angstrom} and 2.1 {angstrom}, respectively. Although the overall fold of the enzyme is similar to that of yeast E3, these two structures differ at two loops that protrude from the proteins and at their FAD-binding sites. The structure of oxidized hE3 with NAD{sup +} bound demonstrates that the nicotinamide moiety is not proximal to the FAD. When NADH is present, however, the nicotinamide base stacks directly on the isoalloxazine ring system of the FAD. This is the first time that this mechanistically requisite conformation of NAD{sup +} or NADH has been observed in E3 from any species. Because E3 structures were previously available only from unicellular organisms, speculations regarding the molecular mechanisms of E3 deficiency were based on homology models. The current hE3 structures show directly that the disease-causing mutations occur at three locations in the human enzyme: the dimer interface, the active site, and the FAD and NAD{sup +}-binding sites. The mechanisms by which these mutations impede the function of hE3 are discussed.

  7. Kinetic and structural basis for acyl-group selectivity and NAD+-dependence in Sirtuin-catalyzed deacylation

    Science.gov (United States)

    Thelen, Julie N.; Ito, Akihiro; Yoshida, Minoru; Denu, John M.

    2015-01-01

    Acylation of lysine is an important protein modification regulating diverse biological processes. It was recently demonstrated that members of the human Sirtuin family are capable of catalyzing long-chain deacylation, in addition to the well-known NAD+-dependent deacetylation activity.1 Here we provide a detailed kinetic and structural analysis that describes the interdependence of NAD+ and acyl-group length for a diverse series of human Sirtuins, SIRT1, SIRT2, SIRT3 and SIRT6. Steady-state and rapid-quench kinetic analyses indicated that differences in NAD+ saturation and susceptibility to nicotinamide inhibition reflect unique kinetic behavior displayed by each Sirtuin and depend on acyl-substrate chain length. Though the rate of nucleophilic attack of the 2′-hydroxyl on the C1′-O-alkylimidate intermediate varies with acyl substrate chain length, this step remains rate-determining for SIRT2 and SIRT3; however for SIRT6, this step is no longer rate-limiting for long-chain substrates. Co-crystallization of SIRT2 with myristoylated peptide and NAD+ yielded a co-complex structure with reaction product 2′-O-myristoyl-ADP-ribose, revealing a latent hydrophobic cavity to accommodate the long chain acyl group, and suggesting a general mechanism for long chain deacylation. Comparing two separately solved co-complex structures containing either a myristoylated peptide or 2′-O-myristoyl-ADP-ribose indicate there are conformational changes at the myristoyl-ribose linkage with minimal structural differences in the enzyme active site. During the deacylation reaction, the fatty acyl group is held in a relatively fixed position. We describe a kinetic and structural model to explain how various Sirtuins display unique acyl-substrate preferences and how different reaction kinetics influence NAD+ dependence. The biological implications are discussed. PMID:25897714

  8. Discovery of a novel inhibitor of NAD(P)(+)-dependent malic enzyme (ME2) by high-throughput screening.

    Science.gov (United States)

    Wen, Yi; Xu, Lei; Chen, Fang-lei; Gao, Jing; Li, Jing-ya; Hu, Li-hong; Li, Jia

    2014-05-01

    Malic enzymes are oxidative decarboxylases with NAD(+) or NAD(P)(+) as cofactor that catalyze the conversion of L-malate to pyruvate and CO2. The aim of this study was to discover and characterize a potent inhibitor of human NAD(P)(+)-dependent malic enzyme 2 (ME2). Recombinant human ME2-His-Tag fusion protein was overexpressed in E coli and purified with Ni-NTA resin. A high-throughput screening (HTS) assay was developed to find ME2 inhibitors. Detergent Brij-35 was used to exclude false positives. The characteristics of the inhibitor were analyzed with enzyme kinetics analysis. A thermal shift assay for ME2 was carried out to verify the binding of the inhibitor with the enzyme. An HTS system for discovering ME2 inhibitors was established with a Z' factor value of 0.775 and a signal-to-noise ratio (S/N) of 9.80. A library containing 12 683 natural products was screened. From 47 hits, NPD387 was identified as an inhibitor of ME2. The primary structure-activity relationship study on NPD387 derivatives showed that one derivative NPD389 was more potent than the parent compound NPD387 (the IC50 of NPD389 was 4.63 ± 0.36 μmol/L or 5.59 ± 0.38 μmol/L, respectively, in the absence or presence of 0.01% Brij-35 in the assay system). The enzyme kinetics analysis showed that NPD389 was a fast-binding uncompetitive inhibitor with respect to the substrate NAD(+) and a mixed-type inhibitor with respect to the substrate L-malate. NPD389 is a potent ME2 inhibitor that binds to the enzyme in a fast-binding mode, acting as an uncompetitive inhibitor with respect to the substrate NAD(+) and a mixed-type inhibitor with respect to the substrate L-malate.

  9. Repeat dose NRPT (nicotinamide riboside and pterostilbene) increases NAD+levels in humans safely and sustainably: a randomized, double-blind, placebo-controlled study.

    Science.gov (United States)

    Dellinger, Ryan W; Santos, Santiago Roel; Morris, Mark; Evans, Mal; Alminana, Dan; Guarente, Leonard; Marcotulli, Eric

    2017-01-01

    NRPT is a combination of nicotinamide riboside (NR), a nicotinamide adenine dinucleotide (NAD +) precursor vitamin found in milk, and pterostilbene (PT), a polyphenol found in blueberries. Here, we report this first-in-humans clinical trial designed to assess the safety and efficacy of a repeat dose of NRPT (commercially known as Basis). NRPT was evaluated in a randomized, double-blind, and placebo-controlled study in a population of 120 healthy adults between the ages of 60 and 80 years. The study consisted of three treatment arms: placebo, recommended dose of NRPT (NRPT 1X), and double dose of NRPT (NRPT 2X). All subjects took their blinded supplement daily for eight weeks. Analysis of NAD + in whole blood demonstrated that NRPT significantly increases the concentration of NAD + in a dose-dependent manner. NAD + levels increased by approximately 40% in the NRPT 1X group and approximately 90% in the NRPT 2X group after 4 weeks as compared to placebo and baseline. Furthermore, this significant increase in NAD + levels was sustained throughout the entire 8-week trial. NAD + levels did not increase for the placebo group during the trial. No serious adverse events were reported in this study. This study shows that a repeat dose of NRPT is a safe and effective way to increase NAD + levels sustainably.

  10. Determination of NAD+ and NADH level in a Single Cell Under H2O2 Stress by Capillary Electrophoresis

    Energy Technology Data Exchange (ETDEWEB)

    Xi, Wenjun [Iowa State Univ., Ames, IA (United States)

    2008-01-01

    A capillary electrophoresis (CE) method is developed to determine both NAD+ and NADH levels in a single cell, based on an enzymatic cycling reaction. The detection limit can reach down to 0.2 amol NAD+ and 1 amol NADH on a home-made CE-LIF setup. The method showed good reproducibility and specificity. After an intact cell was injected into the inlet of a capillary and lysed using a Tesla coil, intracellular NAD+ and NADH were separated, incubated with the cycling buffer, and quantified by the amount of fluorescent product generated. NADH and NAD+ levels of single cells of three cell lines and primary astrocyte culture were determined using this method. Comparing cellular NAD+ and NADH levels with and without exposure to oxidative stress induced by H2O2, it was found that H9c2 cells respond to the stress by reducing both cellular NAD+ and NADH levels, while astrocytes respond by increasing cellular NADH/NAD+ ratio.

  11. Exogenous NAD+ decreases oxidative stress and protects H2O2-treated RPE cells against necrotic death through the up-regulation of autophagy

    Science.gov (United States)

    Zhu, Ying; Zhao, Ke-ke; Tong, Yao; Zhou, Ya-li; Wang, Yi-xiao; Zhao, Pei-quan; Wang, Zhao-yang

    2016-01-01

    Increased oxidative stress, which can lead to the retinal pigment epithelium (RPE) cell death by inducing ATP depletion and DNA repair, is believed to be a prominent pathology in age-related macular degeneration (AMD). In the present study, we showed that and 0.1 mM nicotinamide adenine dinucleotide (NAD+) administration significantly blocked RPE cell death induced by 300 μM H2O2. Further investigation showed that H2O2 resulted in increased intracellular ROS level, activation of PARP-1 and subsequently necrotic death of RPE cells. Exogenous NAD+ administration significantly decreased intracellular and intranuclear ROS levels in H2O2-treated RPE cells. In addition, NAD+ administration to H2O2-treated RPE cells inhibited the activation of PARP-1 and protected the RPE cells against necrotic death. Moreover, exogenous NAD+ administration up-regulated autophagy in the H2O2-treated RPE cells. Inhibition of autophagy by LY294002 blocked the decrease of intracellular and intranuclear ROS level. Besides, inhibition of autophagy by LY294002 abolished the protection of exogenous NAD+ against H2O2-induced cell necrotic death. Taken together, our findings indicate that that exogenous NAD+ administration suppresses H2O2-induced oxidative stress and protects RPE cells against PARP-1 mediated necrotic death through the up-regulation of autophagy. The results suggest that exogenous NAD+ administration might be potential value for the treatment of AMD. PMID:27240523

  12. NAD1 Controls Defense-Like Responses in Medicago truncatula Symbiotic Nitrogen Fixing Nodules Following Rhizobial Colonization in a BacA-Independent Manner.

    Science.gov (United States)

    Domonkos, Ágota; Kovács, Szilárd; Gombár, Anikó; Kiss, Ernő; Horváth, Beatrix; Kováts, Gyöngyi Z; Farkas, Attila; Tóth, Mónika T; Ayaydin, Ferhan; Bóka, Károly; Fodor, Lili; Ratet, Pascal; Kereszt, Attila; Endre, Gabriella; Kaló, Péter

    2017-12-14

    Legumes form endosymbiotic interaction with host compatible rhizobia, resulting in the development of nitrogen-fixing root nodules. Within symbiotic nodules, rhizobia are intracellularly accommodated in plant-derived membrane compartments, termed symbiosomes. In mature nodule, the massively colonized cells tolerate the existence of rhizobia without manifestation of visible defense responses, indicating the suppression of plant immunity in the nodule in the favur of the symbiotic partner. Medicago truncatula DNF2 (defective in nitrogen fixation 2) and NAD1 (nodules with activated defense 1) genes are essential for the control of plant defense during the colonization of the nitrogen-fixing nodule and are required for bacteroid persistence. The previously identified nodule-specific NAD1 gene encodes a protein of unknown function. Herein, we present the analysis of novel NAD1 mutant alleles to better understand the function of NAD1 in the repression of immune responses in symbiotic nodules. By exploiting the advantage of plant double and rhizobial mutants defective in establishing nitrogen-fixing symbiotic interaction, we show that NAD1 functions following the release of rhizobia from the infection threads and colonization of nodule cells. The suppression of plant defense is self-dependent of the differentiation status of the rhizobia. The corresponding phenotype of nad1 and dnf2 mutants and the similarity in the induction of defense-associated genes in both mutants suggest that NAD1 and DNF2 operate close together in the same pathway controlling defense responses in symbiotic nodules.

  13. A NAD-dependent glutamate dehydrogenase coordinates metabolism with cell division in Caulobacter crescentus

    Science.gov (United States)

    Beaufay, François; Coppine, Jérôme; Mayard, Aurélie; Laloux, Géraldine; De Bolle, Xavier; Hallez, Régis

    2015-01-01

    Coupling cell cycle with nutrient availability is a crucial process for all living cells. But how bacteria control cell division according to metabolic supplies remains poorly understood. Here, we describe a molecular mechanism that coordinates central metabolism with cell division in the α-proteobacterium Caulobacter crescentus. This mechanism involves the NAD-dependent glutamate dehydrogenase GdhZ and the oxidoreductase-like KidO. While enzymatically active GdhZ directly interferes with FtsZ polymerization by stimulating its GTPase activity, KidO bound to NADH destabilizes lateral interactions between FtsZ protofilaments. Both GdhZ and KidO share the same regulatory network to concomitantly stimulate the rapid disassembly of the Z-ring, necessary for the subsequent release of progeny cells. Thus, this mechanism illustrates how proteins initially dedicated to metabolism coordinate cell cycle progression with nutrient availability. PMID:25953831

  14. Role of NAD, Oxidative Stress, and Tryptophan Metabolism in Autism Spectrum Disorders

    Directory of Open Access Journals (Sweden)

    Musthafa Mohamed Essa

    2013-01-01

    Full Text Available Autism spectrum disorder (ASD is a pervasive neuro-developmental disorder characterized by impaired social interaction, reduced/absent verbal and non-verbal communication, and repetitive behavior during early childhood. The etiology of this developmental disorder is poorly understood, and no biomarkers have been identified. Identification of novel biochemical markers related to autism would be advantageous for earlier clinical diagnosis and intervention. Studies suggest that oxidative stress-induced mechanisms and reduced antioxidant defense, mitochondrial dysfunction, and impaired energy metabolism (NAD + , NADH, ATP, pyruvate, and lactate, are major causes of ASD. This review provides renewed insight regarding current autism research related to oxidative stress, mitochondrial dysfunction, and altered tryptophan metabolism in ASD.

  15. Two for the Price of One: A Neuroprotective Chaperone Kit within NAD Synthase Protein NMNAT2.

    Directory of Open Access Journals (Sweden)

    Angela Lavado-Roldán

    2016-07-01

    Full Text Available One of the most fascinating properties of the brain is the ability to function smoothly across decades of a lifespan. Neurons are nondividing mature cells specialized in fast electrical and chemical communication at synapses. Often, neurons and synapses operate at high levels of activity through sophisticated arborizations of long axons and dendrites that nevertheless stay healthy throughout years. On the other hand, aging and activity-dependent stress strike onto the protein machineries turning proteins unfolded and prone to form pathological aggregates associated with neurodegeneration. How do neurons protect from those insults and remain healthy for their whole life? Ali and colleagues now present a molecular mechanism by which the enzyme nicotinamide mononucleotide adenylyltransferase 2 (NMNAT2 acts not only as a NAD synthase involved in axonal maintenance but as a molecular chaperone helping neurons to overcome protein unfolding and protein aggregation.

  16. Mechanism of NAD(P)H:quinone reductase: Ab initio studies of reduced flavin.

    Science.gov (United States)

    Cavelier, G; Amzel, L M

    2001-06-01

    NAD(P)H:quinone oxidoreductase type 1 (QR1, NQO1, formerly DT-diaphorase; EC 1.6.99.2) is an FAD-containing enzyme that catalyzes the nicotinamide nucleotide-dependent reduction of quinones, quinoneimines, azo dyes, and nitro groups. Animal cells are protected by QR1 from the toxic and neoplastic effects of quinones and other electrophiles. Alternatively, in tumor cells QR can activate a number of cancer chemotherapeutic agents such as mitomycins and aziridylbenzoquinones. Thus, the same enzyme that protects the organism from the deleterious effects of quinones can activate cytotoxic chemotherapeutic prodrugs and cause cancer cell death. The catalytic mechanism of QR includes an important initial step in which FAD is reduced by NAD(P)H. The unfavorable charge separation that results must be stabilized by the protein. The details of this charge stabilization step are inaccessible to easy experimental verification but can be studied by quantum chemistry methods. Here we report ab initio quantum mechanical calculations in and around the active site of the enzyme that provide information about the fine details of the contribution of the protein to the stabilization of the reduced flavin. The results show that (1) protein interactions provide approximately 2 kcal/mol to stabilize the planar conformation of the reduced flavin isoalloxazine ring observed in the X-ray structure; (2) the charge separation present in the reduced planar form of the flavin is stabilized by interactions with groups of the protein; (3) even after stabilization, the reduction potential of the cofactor remains more negative than that of the free flavin, making it a better reductant for a larger variety of quinones; and (4) the more negative reduction potential may also result in faster kinetics for the quinone reduction step. Copyright 2001 Wiley-Liss, Inc.

  17. Variation in Streptococcus pyogenes NAD+ glycohydrolase is associated with tissue tropism.

    Science.gov (United States)

    Riddle, David J; Bessen, Debra E; Caparon, Michael G

    2010-07-01

    Streptococcus pyogenes is an important pathogen that causes a variety of diseases. The most common infections involve the throat (pharyngitis) or skin (impetigo); however, the factors that determine tissue tropism and severity are incompletely understood. The S. pyogenes NAD(+) glycohydrolase (SPN) is a virulence factor that has been implicated in contributing to the pathogenesis of severe infections. However, the role of SPN in determining the bacterium's tissue tropism has not been evaluated. In this report, we examine the sequences of spn and its endogenous inhibitor ifs from a worldwide collection of S. pyogenes strains. Analysis of average pairwise nucleotide diversity, average number of nucleotide differences, and ratio of nonsynonymous to synonymous substitutions revealed significant diversity in spn and ifs. Application of established models of molecular evolution shows that SPN is evolving under positive selection and diverging into NAD(+) glycohydrolase (NADase)-active and -inactive subtypes. Additionally, the NADase-inactive SPN subtypes maintain the characteristics of a functional gene while ifs becomes a pseudogene. Thus, NADase-inactive SPN continues to evolve under functional constraint. Furthermore, NADase activity did not correlate with invasive disease in our collection but was associated with tissue tropism. The ability to cause infection at both the pharynx and the skin ("generalist" strains) is correlated with NADase-active SPN, while the preference for causing infection at either the throat or the skin ("specialist" strains) is associated with NADase-inactive SPN. These findings suggest that SPN has a NADase-independent function and prompt a reevaluation of the role of SPN in streptococcal pathogenesis.

  18. Angular distributions of autoionization electrons from Ne(2p43s2) 1D in Li+-Ne collisions

    International Nuclear Information System (INIS)

    Oud, M.; Pas, S.F. te; Westerveld, W.B.; Niehaus, A.

    1993-01-01

    Angular distributions of autoionization electrons from Ne(2p 4 3s 2 ) 1 D due to Li + -Ne collisions measured in coincidence with the scattered projectile ions are presented. The measurements are performed at four different collision energies between 1.0 keV and 3.0 keV, and the complex population amplitudes for the excited 1 D state are determined. A nearly pure M = O sublevel population is found with respect to an axis coinciding with the direction of the angular distribution. The direction of the angular distribution is found to deviate from the final direction of the asymptotic internuclear axis. (author)

  19. Acute Ethanol Intake Induces NAD(P)H Oxidase Activation and Rhoa Translocation in Resistance Arteries.

    Science.gov (United States)

    Simplicio, Janaina A; Hipólito, Ulisses Vilela; Vale, Gabriel Tavares do; Callera, Glaucia Elena; Pereira, Camila André; Touyz, Rhian M; Tostes, Rita de Cássia; Tirapelli, Carlos R

    2016-11-01

    The mechanism underlying the vascular dysfunction induced by ethanol is not totally understood. Identification of biochemical/molecular mechanisms that could explain such effects is warranted. To investigate whether acute ethanol intake activates the vascular RhoA/Rho kinase pathway in resistance arteries and the role of NAD(P)H oxidase-derived reactive oxygen species (ROS) on such response. We also evaluated the requirement of p47phox translocation for ethanol-induced NAD(P)H oxidase activation. Male Wistar rats were orally treated with ethanol (1g/kg, p.o. gavage) or water (control). Some rats were treated with vitamin C (250 mg/kg, p.o. gavage, 5 days) before administration of water or ethanol. The mesenteric arterial bed (MAB) was collected 30 min after ethanol administration. Vitamin C prevented ethanol-induced increase in superoxide anion (O2-) generation and lipoperoxidation in the MAB. Catalase and superoxide dismutase activities and the reduced glutathione, nitrate and hydrogen peroxide (H2O2) levels were not affected by ethanol. Vitamin C and 4-methylpyrazole prevented the increase on O2- generation induced by ethanol in cultured MAB vascular smooth muscle cells. Ethanol had no effect on phosphorylation levels of protein kinase B (Akt) and eNOS (Ser1177 or Thr495 residues) or MAB vascular reactivity. Vitamin C prevented ethanol-induced increase in the membrane: cytosol fraction ratio of p47phox and RhoA expression in the rat MAB. Acute ethanol intake induces activation of the RhoA/Rho kinase pathway by a mechanism that involves ROS generation. In resistance arteries, ethanol activates NAD(P)H oxidase by inducing p47phox translocation by a redox-sensitive mechanism. O mecanismo da disfunção vascular induzido pelo consumo de etanol não é totalmente compreendido. Justifica-se, assim a identificação de mecanismos bioquímicos e moleculares que poderiam explicar tais efeitos. Investigar se a ingestão aguda de etanol ativa a via vascular RhoA/Rho quinase

  20. On the recovery of the DNA-synthesis after X-irradiation in the spleen of mice and its modification by the NAD-metabolism

    International Nuclear Information System (INIS)

    Streffer, C.

    1974-01-01

    The incorporation of tritium-labelled thymidine into the DNA of mice spleen cells after whole body irradiation with X-rays was measured in order to study the decrease of DNA synthesis is decreased for several hours after irradiation with low doses. Recovery effects become operative after six hours. The radiation effect on the NAD metabolism, known to be related to DNA synthesis, was also investigated. The rate of NAD synthesis is influenced via the extremely radiosensitive metabolic process in the nucleus. Conversely, inhibition of DNA synthesis by injection of NAD enhances the recovery of DNA synthesis after irradiaton. (G.G.)

  1. Status of conversion of NE standards to national consensus standards

    International Nuclear Information System (INIS)

    Jennings, S.D.

    1990-06-01

    One major goal of the Nuclear Standards Program is to convert existing NE standards into national consensus standards (where possible). This means that an NE standard in the same subject area using the national consensus process. This report is a summary of the activities that have evolved to effect conversion of NE standards to national consensus standards, and the status of current conversion activities. In some cases, all requirements in an NE standard will not be incorporated into the published national consensus standard because these requirements may be considered too restrictive or too specific for broader application by the nuclear industry. If these requirements are considered necessary for nuclear reactor program applications, the program standard will be revised and issued as a supplement to the national consensus standard. The supplemental program standard will contain only those necessary requirements not reflected by the national consensus standard. Therefore, while complete conversion of program standards may not always be realized, the standards policy has been fully supported in attempting to make maximum use of the national consensus standard. 1 tab

  2. Stress tolerant plant species spread in the road-ne

    Czech Academy of Sciences Publication Activity Database

    Šerá, Božena

    2011-01-01

    Roč. 14, Vol.14 (2011), s. 45-46 ISSN 1644-7298 R&D Projects: GA MŠk OC10032 Institutional research plan: CEZ:AV0Z60870520 Keywords : weed * invasive * road -ne * salinity * Poaceae Subject RIV: AP - Urban, Regional and Transport Planning

  3. "Dlja menja ne sushtshestvujut kraski..." : [luuletused] / Georgi Kirillov

    Index Scriptorium Estoniae

    Kirillov, Georgi, 1952-2016

    2002-01-01

    Autorist lk. 187. Sisu: "Dlja menja ne sushtshestvujut kraski..." ; "Za molitvoi molitva..." ; "Zaklannõi prezhde veka Agnets..." ; "Ja zhdal tebja i tõ voshol..." ; "Nepodrazhajemoje solntse..." ; "Tshto obshtshego mezh mnoju i toboi..." ; "Pogruzhenije v odinotshestvo..." ; "Shag za shagom - k stupenjam svjatõm..." ; "Nedvizhnõ dveri sozertsanja..." ; "Vessenni vozduh..." ; "Jesli mozhno - bud miloserdnõm..."

  4. 76 FR 76337 - Television Broadcasting Services; Lincoln, NE

    Science.gov (United States)

    2011-12-07

    ... FEDERAL COMMUNICATIONS COMMISSION 47 CFR Part 73 [MB Docket No. 11-192, RM-11646; DA 11-1924] Television Broadcasting Services; Lincoln, NE AGENCY: Federal Communications Commission. ACTION: Proposed... 73 Television, Television broadcasting. Federal Communications Commission Barbara A. Kreisman, Chief...

  5. Hyurterianum (Asteraceae, Inuleae), a new species from NE Anatolia, Turkey

    DEFF Research Database (Denmark)

    Gemici, Y.; Tan, K.; Yidirim, H.

    2008-01-01

    Helichrysum yurterianum Y. Gemici, Kit Tan, H. Yildirim & M. Gemici (Asteraceae, Inuleae) is described and illustrated. It is a serpentine endemic restricted to the province of Erzincan in NE Anatolia, Turkey. Its affinities are with H. arenarium and H. noeanum, which both have a wider distribution...

  6. Ceux-ci ne sont pas : [luuletused] / Kalju Kruusa

    Index Scriptorium Estoniae

    Kruusa, Kalju, pseud., 1973-

    2003-01-01

    Sisu: Ceux-ci ne sont pas ; Köögivahet ; "Taara..." ; "Pakane pistab pisikesi ..." ; "Meri on kaet mattklaasiga ; Pydemise päivil ; "Toas muusika mängib ..." ; "Jäin juustu imetlema ; "Mu elu on mustikas ..." ; Hingepidetus ; ŁNo me gusta la cocina

  7. HeNe-laser light scattering by human dental enamel

    NARCIS (Netherlands)

    Zijp, [No Value; tenBosch, JJ; Groenhuis, RAJ

    1995-01-01

    Knowledge of the optical properties of tooth enamel and an understanding of the origin of these properties are necessary for the development of new optical methods for caries diagnosis and the measurement of tooth color. We measured the scattering intensity functions for HeNe-laser light of 80- to

  8. Wiiralti kaasaegsed lääne graafikas / Mai Levin

    Index Scriptorium Estoniae

    Levin, Mai, 1942-

    2003-01-01

    Adamson-Ericu muuseumis näitus "Wiiralti kaasaegsed lääne graafikas", mis on koostatud Nina Poomi (sünd. 1909, H. Radevalli ema) kogusse kuulunud töödest. Näitus tutvustab E. Wiiralti sõjajärgse loomingu tausta, jäädvustab kunstikogu olemasolu

  9. Recoil range distribution measurement in 20Ne + 181Ta reaction

    International Nuclear Information System (INIS)

    Tripathi, R.; Sudarshan, K.; Goswami, A.; Guin, R.; Reddy, A.V.R.

    2005-01-01

    In order to investigate linear momentum transfer in various transfer channels in 20 Ne + 181 Ta, recoil range distribution measurements have been carried out at E lab = 180 MeV, populating significant number of l-waves above l crit

  10. Deformation effects in the 20Ne+12C reaction

    International Nuclear Information System (INIS)

    Dey, A.; Bhattacharya, C.; Banerjee, K.; Kundu, S.; Mukhopadhyay, S.; Gupta, D.; Saha, R.; Bhattacharya, S.

    2004-01-01

    The present work has been performed with the aim to investigate the possible occurrence of highly deformed configurations of the 32 S di-nuclear systems which may be formed in the 20 Ne+ 12 C reaction by studying the properties of emitted light charged particles

  11. Precision angle-resolved autoionization resonances in Ar and Ne

    Energy Technology Data Exchange (ETDEWEB)

    Berrah, N.; Langer, B.; Gorczyca, T.W. [Western Michigan Univ., Kalamazoo, MI (United States)] [and others

    1997-04-01

    Theoretical work has shown that the electron angular distribution and the shape of the autoionization resonances are crucial to the understanding of certain types of electron-electron correlation. Autoionization resonances in Ne (Ar) result from the decay of the excited discrete state Ne{sup *} 2s2p{sup 6} np (Ar{sup *} 3s3p{sup 6} np) into the continuum state Ne{sup +} 2s{sup 2}2p{sup 5} + e{sup {minus}} (ks,kd) (Ar{sup +} 3s{sup 2}3p{sup 5} + e{sup {minus}} (ks,kd)). Since the continuum can also be reached by direct photoionization, both paths add coherently, giving rise to interferences that produce the characteristic Beutler-Fano line shape. In this work, the authors report on quantitative angle-resolved electron spectrometry studies of (a) the Ne 2s{sup 2}2p{sup 6} {r_arrow} 2s2p{sup 6} np (n=3-5) autoionizing resonances and the 2s{sup 2}2p{sup 6} {r_arrow} 2p{sup 4}3s3p doubly excited resonance, (b) the Ar 3s{sup 2}3p{sup 6} {r_arrow} 3s3p{sup 6} np (n=4-9) autoionization resonances and extended R-matrix calculations of the angular-distribution parameters for both Ne and Ar measurements. Their results are compared with previous theoretical work by Taylor.

  12. Localization of inner-shell photoelectron emission and ICD in Ne{sub 2}

    Energy Technology Data Exchange (ETDEWEB)

    Kreidi, K; Jahnke, T; Havermeier, T; Grisenti, R E; Schoessler, S; Schmidt, L Ph H; Schoeffler, M; Odenweller, M; Neumann, N; Foucar, L; Titze, J; Ulrich, B; Sturm, F; Stuck, C; Wallauer, R; Voss, S; Lauter, I [Institut fuer Kernphysik, J. W. Goethe Universitaet, Max-von-Laue-Str. 1, 60438 Frankfurt (Germany); Weber, T H [Lawrence Berkeley National Laboratory, Berkeley CA 94720 (United States); Liu, X [Institute of Multidisciplinary Research for Advanced Materials, Tohoku University, Sendai 980-8577 (Japan); Morisita, Y, E-mail: kreidi@atom.uni-frankfurt.d [National Metrology Institute of Japan, AIST, Tsukuba 305-8568 (Japan)

    2009-11-01

    With photon energies 10 eV above the K-edge the 1s photoionization of neon dimers was measured by using COLd Target Recoil Ion Momentum Spectroscopy. All decay channels resulting in the breakup Ne{sup +} + Ne{sup +} and Ne{sup 2+} + Ne{sup +}were identified. The latter was used to investigate the localization or delocalization of vacancies in the homonuclear diatomic system.

  13. Measurements of Neutrino Charged Current Interactions at SciBooNE

    Energy Technology Data Exchange (ETDEWEB)

    Nakajima, Yasuhiro [Department of Physics, Kyoto University, Kyoto 606-8502 (Japan)], E-mail: nakajima@scphys.kyoto-u.ac.jp

    2009-08-15

    The SciBooNE experiment (FNAL-E954) is designed to measure neutrino-nucleous cross sections in the one GeV region. Additionally, SciBooNE serves as a near detector for MiniBooNE by measuring the neutrino flux. In this paper, we describe two analyses using neutrino charged current interactions at SciBooNE: a neutrino spectrum measurement and a search for charged current coherent pion production.

  14. [Activity of liver mitochondrial NAD+-dependent dehydrogenases of the krebs cycle in rats with acetaminophen-induced hepatitis developed under conditions of alimentary protein deficiency].

    Science.gov (United States)

    Voloshchuk, O N; Kopylchuk, G P

    2016-01-01

    Activity of isocitrate dehydrogenase, α-ketoglutarate dehydrogenase, malate dehydrogenase, and the NAD(+)/NADН ratio were studied in the liver mitochondrial fraction of rats with toxic hepatitis induced by acetaminophen under conditions of alimentary protein deprivation. Acetaminophen-induced hepatitis was characterized by a decrease of isocitrate dehydrogenase, α-ketoglutarate dehydrogenase and malate dehydrogenase activities, while the mitochondrial NAD(+)/NADН ratio remained at the control level. Modeling of acetaminophen-induced hepatitis in rats with alimentary protein caused a more pronounced decrease in the activity of NAD(+)-dependent dehydrogenases studied and a 2.2-fold increase of the mitochondrial NAD(+)/NADН ratio. This suggests that alimentary protein deprivation potentiated drug-induced liver damage.

  15. Fabrication of Flexible Arrayed Lactate Biosensor Based on Immobilizing LDH-NAD+ on NiO Film Modified by GO and MBs

    Directory of Open Access Journals (Sweden)

    Jung-Chuan Chou

    2017-07-01

    Full Text Available We proposed the flexible arrayed lactate biosensor based on immobilizing l-lactate dehydrogenase (LDH and nicotinamide adenine dinucleotide ( NAD + on nickel oxide (NiO film, and which the average sensitivity could be enhanced by using graphene oxide (GO and magnetic beads (MBs. By using GO and MBs, it exhibits excellent sensitivity (45.397 mV/mM with a linearity of 0.992 in a range of 0.2 mM to 3 mM. According to the results of electrochemical impedance spectroscopy (EIS, the electron transfer resistance of LDH- NAD + -MBs/GPTS/GO/NiO film was smaller than those of LDH-NAD+/GPTS/GO/NiO film and LDH- NAD + /GPTS/NiO film, and it presented the outstanding electron transfer ability. After that, the limit of detection, anti-interference effect and bending test were also investigated.

  16. Koliesko - studentský pracovní seminář při festivalu lidové kultury v Kokavě nad Rimavicou

    Czech Academy of Sciences Publication Activity Database

    Císaríková, Klára; Maňáková, M.

    2009-01-01

    Roč. 68, č. 2 (2009), 107-110 ISSN 1211-8117 Institutional research plan: CEZ:AV0Z90580513 Keywords : folk culture * Kokava nad Rimavicou * student´s workshop Subject RIV: AC - Archeology, Anthropology, Ethnology

  17. Purification and characterization of an NAD+-linked formaldehyde dehydrogenase from the facultative RuMP cycle methylotroph Arthrobacter P1

    NARCIS (Netherlands)

    Attwood, Margaret M.; Arfman, Nico; Weusthuis, Ruud A.; Dijkhuizen, Lubbert

    1992-01-01

    When Arthrobacter P1 is grown on choline, betaine, dimethylglycine or sarcosine, an NAD+-dependent formaldehyde dehydrogenase is induced. This formaldehyde dehydrogenase has been purified using ammonium sulphate fractionation, anion exchange- and hydrophobic interaction chromatography. The molecular

  18. 4-Pyridone-3-carboxamide-1-β-D-ribonucleoside triphosphate (4PyTP), a novel NAD metabolite accumulating in erythrocytes of uremic children: a biomarker for a toxic NAD analogue in other tissues?

    Science.gov (United States)

    Synesiou, Elena; Fairbanks, Lynnette D; Simmonds, H Anne; Slominska, Ewa M; Smolenski, Ryszard T; Carrey, Elizabeth A

    2011-06-01

    We have identified a novel nucleotide, 4-pyridone 3/5-carboxamide ribonucleoside triphosphate (4PyTP), which accumulates in human erythrocytes during renal failure. Using plasma and erythrocyte extracts obtained from children with chronic renal failure we show that the concentration of 4PyTP is increased, as well as other soluble NAD(+) metabolites (nicotinamide, N(1)-methylnicotinamide and 4Py-riboside) and the major nicotinamide metabolite N(1)-methyl-2-pyridone-5-carboxamide (2PY), with increasing degrees of renal failure. We noted that 2PY concentration was highest in the plasma of haemodialysis patients, while 4PyTP was highest in erythrocytes of children undergoing peritoneal dialysis: its concentration correlated closely with 4Py-riboside, an authentic precursor of 4PyTP, in the plasma. In the dialysis patients, GTP concentration was elevated: similar accumulation was noted previously, as a paradoxical effect in erythrocytes during treatment with immunosuppressants such as ribavirin and mycophenolate mofetil, which deplete GTP through inhibition of IMP dehydrogenase in nucleated cells such as lymphocytes. We predict that 4Py-riboside and 4Py-nucleotides bind to this enzyme and alter its activity. The enzymes that regenerate NAD(+) from nicotinamide riboside also convert the drugs tiazofurin and benzamide riboside into NAD(+) analogues that inhibit IMP dehydrogenase more effectively than the related ribosides: we therefore propose that the accumulation of 4PyTP in erythrocytes during renal failure is a marker for the accumulation of a related toxic NAD(+) analogue that inhibits IMP dehydrogenase in other cells.

  19. 4-Pyridone-3-carboxamide-1-β-d-ribonucleoside Triphosphate (4PyTP, a Novel NAD+ Metabolite Accumulating in Erythrocytes of Uremic Children: A Biomarker for a Toxic NAD+ Analogue in Other Tissues?

    Directory of Open Access Journals (Sweden)

    Elizabeth A. Carrey

    2011-06-01

    Full Text Available We have identified a novel nucleotide, 4-pyridone 3/5-carboxamide ribonucleoside triphosphate (4PyTP, which accumulates in human erythrocytes during renal failure. Using plasma and erythrocyte extracts obtained from children with chronic renal failure we show that the concentration of 4PyTP is increased, as well as other soluble NAD+ metabolites (nicotinamide, N1-methylnicotinamide and 4Py-riboside and the major nicotinamide metabolite N1-methyl-2-pyridone-5-carboxamide (2PY, with increasing degrees of renal failure. We noted that 2PY concentration was highest in the plasma of haemodialysis patients, while 4PyTP was highest in erythrocytes of children undergoing peritoneal dialysis: its concentration correlated closely with 4Py-riboside, an authentic precursor of 4PyTP, in the plasma. In the dialysis patients, GTP concentration was elevated: similar accumulation was noted previously, as a paradoxical effect in erythrocytes during treatment with immunosuppressants such as ribavirin and mycophenolate mofetil, which deplete GTP through inhibition of IMP dehydrogenase in nucleated cells such as lymphocytes. We predict that 4Py-riboside and 4Py-nucleotides bind to this enzyme and alter its activity. The enzymes that regenerate NAD+ from nicotinamide riboside also convert the drugs tiazofurin and benzamide riboside into NAD+ analogues that inhibit IMP dehydrogenase more effectively than the related ribosides: we therefore propose that the accumulation of 4PyTP in erythrocytes during renal failure is a marker for the accumulation of a related toxic NAD+ analogue that inhibits IMP dehydrogenase in other cells.

  20. An open-label, non-randomized study of the pharmacokinetics of the nutritional supplement nicotinamide riboside (NR and its effects on blood NAD+ levels in healthy volunteers.

    Directory of Open Access Journals (Sweden)

    Sophia E Airhart

    Full Text Available The co-primary objectives of this study were to determine the human pharmacokinetics (PK of oral NR and the effect of NR on whole blood nicotinamide adenine dinucleotide (NAD+ levels.Though mitochondrial dysfunction plays a critical role in the development and progression of heart failure, no mitochondria-targeted therapies have been translated into clinical practice. Recent murine studies have reported associations between imbalances in the NADH/NAD+ ratio with mitochondrial dysfunction in multiple tissues, including myocardium. Moreover, an NAD+ precursor, nicotinamide mononucleotide, improved cardiac function, while another NAD+ precursor, nicotinamide riboside (NR, improved mitochondrial function in muscle, liver and brown adipose. Thus, PK studies of NR in humans is critical for future clinical trials.In this non-randomized, open-label PK study of 8 healthy volunteers, 250 mg NR was orally administered on Days 1 and 2, then uptitrated to peak dose of 1000 mg twice daily on Days 7 and 8. On the morning of Day 9, subjects completed a 24-hour PK study after receiving 1000 mg NR at t = 0. Whole-blood levels of NR, clinical blood chemistry, and NAD+ levels were analyzed.Oral NR was well tolerated with no adverse events. Significant increases comparing baseline to mean concentrations at steady state (Cave,ss were observed for both NR (p = 0.03 and NAD+ (p = 0.001; the latter increased by 100%. Absolute changes from baseline to Day 9 in NR and NAD+ levels correlated highly (R2 = 0.72, p = 0.008.Because NR increases circulating NAD+ in humans, NR may have potential as a therapy in patients with mitochondrial dysfunction due to genetic and/or acquired diseases.

  1. An open-label, non-randomized study of the pharmacokinetics of the nutritional supplement nicotinamide riboside (NR) and its effects on blood NAD+ levels in healthy volunteers.

    Science.gov (United States)

    Airhart, Sophia E; Shireman, Laura M; Risler, Linda J; Anderson, Gail D; Nagana Gowda, G A; Raftery, Daniel; Tian, Rong; Shen, Danny D; O'Brien, Kevin D

    2017-01-01

    The co-primary objectives of this study were to determine the human pharmacokinetics (PK) of oral NR and the effect of NR on whole blood nicotinamide adenine dinucleotide (NAD+) levels. Though mitochondrial dysfunction plays a critical role in the development and progression of heart failure, no mitochondria-targeted therapies have been translated into clinical practice. Recent murine studies have reported associations between imbalances in the NADH/NAD+ ratio with mitochondrial dysfunction in multiple tissues, including myocardium. Moreover, an NAD+ precursor, nicotinamide mononucleotide, improved cardiac function, while another NAD+ precursor, nicotinamide riboside (NR), improved mitochondrial function in muscle, liver and brown adipose. Thus, PK studies of NR in humans is critical for future clinical trials. In this non-randomized, open-label PK study of 8 healthy volunteers, 250 mg NR was orally administered on Days 1 and 2, then uptitrated to peak dose of 1000 mg twice daily on Days 7 and 8. On the morning of Day 9, subjects completed a 24-hour PK study after receiving 1000 mg NR at t = 0. Whole-blood levels of NR, clinical blood chemistry, and NAD+ levels were analyzed. Oral NR was well tolerated with no adverse events. Significant increases comparing baseline to mean concentrations at steady state (Cave,ss) were observed for both NR (p = 0.03) and NAD+ (p = 0.001); the latter increased by 100%. Absolute changes from baseline to Day 9 in NR and NAD+ levels correlated highly (R2 = 0.72, p = 0.008). Because NR increases circulating NAD+ in humans, NR may have potential as a therapy in patients with mitochondrial dysfunction due to genetic and/or acquired diseases.

  2. Coenzymatic properties of low molecular-weight and macromolecular N6-derivatives of NAD+ and NADP+ with dehydrogenases of interest for organic synthesis.

    Science.gov (United States)

    Ottolina, G; Carrea, G; Riva, S; Bückmann, A F

    1990-08-01

    The catalytic activity, expressed as Km and Vmax values, of 16 enzymes of practical interest with the macromolecular coenzymes poly(ethylene glycol)-N6-(2-aminoethyl)-NAD+ and poly(ethylene glycol)-N6-(2-aminoethyl)-NADP+ and their low molecular weight precursors N6-(2-aminoethyl)-NAD+ and N6-(2-aminoethyl)-NADP+, was investigated. The enzymes examined are of direct interest for organic synthesis (i.e. alcohol dehydrogenase from yeast, horse liver, or Thermoanaerobium brockii, lactic dehydrogenase, and several hydroxysteroid dehydrogenases) or are used for the regeneration of NAD+, NADP+, NADH, or NADPH (i.e. glutamate dehydrogenase from liver or Proteus, formate dehydrogenase, glucose dehydrogenase, and malic enzyme). The cycling efficiency of poly(ethylene glycol)-N6-(2-aminoethyl)-NADP+ was examined with coupled-enzymes or coupled-substrates systems. Poly(ethylene glycol)-N6-(2-aminoethyl)-NAD+ and, even more so, poly(ethylene glycol)-N6-(2-aminoethyl)-NADP+ were excellent coenzymes with several dehydrogenases. In addition, the coenzymatic properties of N6-(3-sulfonatopropyl)-NAD+, an NAD+ derivative carrying a strong anionic group, were compared with those of the newly synthesized N6-(2-hydroxy-3-trimethylammonium propyl)-NAD+, an NAD+ derivative carrying a strong cationic group. It was expected that the presence of the sulfonic or quaternary ammonium group would enhance the residence time of the coenzyme inside continuous-flow reactors if membranes with anionic or cationic groups, respectively, were used.

  3. Wld(S reduces paraquat-induced cytotoxicity via SIRT1 in non-neuronal cells by attenuating the depletion of NAD.

    Directory of Open Access Journals (Sweden)

    Qiujing Yu

    Full Text Available Wld(S is a fusion protein with NAD synthesis activity, and has been reported to protect axonal and synaptic compartments of neurons from various mechanical, genetic and chemical insults. However, whether Wld(S can protect non-neuronal cells against toxic chemicals is largely unknown. Here we found that Wld(S significantly reduced the cytotoxicity of bipyridylium herbicides paraquat and diquat in mouse embryonic fibroblasts, but had no effect on the cytotoxicity induced by chromium (VI, hydrogen peroxide, etoposide, tunicamycin or brefeldin A. Wld(S also slowed down the death of mice induced by intraperitoneal injection of paraquat. Further studies demonstrated that Wld(S markedly attenuated mitochondrial injury including disruption of mitochondrial membrane potential, structural damage and decline of ATP induced by paraquat. Disruption of the NAD synthesis activity of Wld(S by an H112A or F116S point mutation resulted in loss of its protective function against paraquat-induced cell death. Furthermore, Wld(S delayed the decrease of intracellular NAD levels induced by paraquat. Similarly, treatment with NAD or its precursor nicotinamide mononucleotide attenuated paraquat-induced cytotoxicity and decline of ATP and NAD levels. In addition, we showed that SIRT1 was required for both exogenous NAD and Wld(S-mediated cellular protection against paraquat. These findings suggest that NAD and SIRT1 mediate the protective function of Wld(S against the cytotoxicity induced by paraquat, which provides new clues for the mechanisms underlying the protective function of Wld(S in both neuronal and non-neuronal cells, and implies that attenuation of NAD depletion may be effective to alleviate paraquat poisoning.

  4. WldS Reduces Paraquat-Induced Cytotoxicity via SIRT1 in Non-Neuronal Cells by Attenuating the Depletion of NAD

    Science.gov (United States)

    Yu, Qiujing; Wang, Ting; Zhou, Xuexia; Wu, Jingxia; Chen, Xingmiao; Liu, Yang; Wu, Dongmei; Zhai, Qiwei

    2011-01-01

    WldS is a fusion protein with NAD synthesis activity, and has been reported to protect axonal and synaptic compartments of neurons from various mechanical, genetic and chemical insults. However, whether WldS can protect non-neuronal cells against toxic chemicals is largely unknown. Here we found that WldS significantly reduced the cytotoxicity of bipyridylium herbicides paraquat and diquat in mouse embryonic fibroblasts, but had no effect on the cytotoxicity induced by chromium (VI), hydrogen peroxide, etoposide, tunicamycin or brefeldin A. WldS also slowed down the death of mice induced by intraperitoneal injection of paraquat. Further studies demonstrated that WldS markedly attenuated mitochondrial injury including disruption of mitochondrial membrane potential, structural damage and decline of ATP induced by paraquat. Disruption of the NAD synthesis activity of WldS by an H112A or F116S point mutation resulted in loss of its protective function against paraquat-induced cell death. Furthermore, WldS delayed the decrease of intracellular NAD levels induced by paraquat. Similarly, treatment with NAD or its precursor nicotinamide mononucleotide attenuated paraquat-induced cytotoxicity and decline of ATP and NAD levels. In addition, we showed that SIRT1 was required for both exogenous NAD and WldS-mediated cellular protection against paraquat. These findings suggest that NAD and SIRT1 mediate the protective function of WldS against the cytotoxicity induced by paraquat, which provides new clues for the mechanisms underlying the protective function of WldS in both neuronal and non-neuronal cells, and implies that attenuation of NAD depletion may be effective to alleviate paraquat poisoning. PMID:21750730

  5. A small-molecule inhibitor suppresses the tumor-associated mitochondrial NAD(P)+-dependent malic enzyme (ME2) and induces cellular senescence.

    Science.gov (United States)

    Hsieh, Ju-Yi; Li, Shao-Yu; Tsai, Wen-Chen; Liu, Jyung-Hurng; Lin, Chih-Li; Liu, Guang-Yaw; Hung, Hui-Chih

    2015-08-21

    Here, we found a natural compound, embonic acid (EA), that can specifically inhibit the enzymatic activity of mitochondrial NAD(P)+-dependent malic enzyme (m-NAD(P)-ME, ME2) either in vitro or in vivo. The in vitro IC50 value of EA for m-NAD(P)-ME was 1.4 ± 0.4 μM. Mutagenesis and binding studies revealed that the putative binding site of EA on m-NAD(P)-ME is located at the fumarate binding site or at the dimer interface near the site. Inhibition studies reveal that EA displayed a non-competitive inhibition pattern, which demonstrated that the binding site of EA was distinct from the active site of the enzyme. Therefore, EA is thought to be an allosteric inhibitor of m-NAD(P)-ME. Both EA treatment and knockdown of m-NAD(P)-ME by shRNA inhibited the growth of H1299 cancer cells. The protein expression and mRNA synthesis of m-NAD(P)-ME in H1299 cells were not influenced by EA, suggesting that the EA-inhibited H1299 cell growth occurs through the suppression of in vivo m-NAD(P)-ME activity EA treatment further induced the cellular senescence of H1299 cells. However, down-regulation of the enzyme-induced cellular senescence was not through p53. Therefore, the EA-evoked senescence of H1299 cells may occur directly through the inhibition of ME2 or a p53-independent pathway.

  6. Crystal structures of complexes of NAD+-dependent formate dehydrogenase from methylotrophic bacterium Pseudomonas sp. 101 with formate

    International Nuclear Information System (INIS)

    Filippova, E. V.; Polyakov, K. M.; Tikhonova, T. V.; Stekhanova, T. N.; Boiko, K. M.; Sadykhov, I. G.; Tishkov, V. I.; Popov, V. O.; Labru, N.

    2006-01-01

    Formate dehydrogenase (FDH) from the methylotrophic bacterium Pseudomonas sp. 101 catalyzes oxidation of formate to NI 2 with the coupled reduction of nicotinamide adenine dinucleotide (NAD + ). The three-dimensional structures of the apo form (the free enzyme) and the holo form (the ternary FDH-NAD + -azide complex) of FDH have been established earlier. In the present study, the structures of FDH complexes with formate are solved at 2.19 and 2.28 A resolution by the molecular replacement method and refined to the R factors of 22.3 and 20.5%, respectively. Both crystal structures contain four protein molecules per asymmetric unit. These molecules form two dimers identical to the dimer of the apo form of FDH. Two possible formatebinding sites are found in the active site of the FDH structure. In the complexes the sulfur atom of residue Cys354 exists in the oxidized state

  7. A high-fat diet and NAD+ activate sirt1 to rescue premature aging in cockayne syndrome

    DEFF Research Database (Denmark)

    Scheibye-Knudsen, Morten; Mitchell, Sarah J.; Fang, Evandro F.

    2014-01-01

    SIRT1 activity and mitochondrial dysfunction. Notably, β-hydroxybutyrate levels are increased by the high-fat diet, and β-hydroxybutyrate, PARP inhibition, or NAD+ supplementation can activate SIRT1 and rescue CS-associated phenotypes. Mechanistically, CSB can displace activated PARP1 from damaged DNA......-fat, caloric-restricted, or resveratrol-supplemented diet. High-fat feeding rescued the metabolic, transcriptomic, and behavioral phenotypes of Csbm/m mice. Furthermore, premature aging in CS mice, nematodes, and human cells results from aberrant PARP activation due to deficient DNA repair leading to decreased...... to limit its activity. This study connects two emerging longevity metabolites, β-hydroxybutyrate and NAD+, through the deacetylase SIRT1 and suggests possible interventions for CS....

  8. Wireless Telegraphy at the German Universal Exhibition in Ústí nad Labem in 1903 [1

    Directory of Open Access Journals (Sweden)

    T. Okurka

    2008-01-01

    Full Text Available This paper focuses on the transmission of wireless telegraphy between Ústí nad Labem and Teplice during the German Universal Exhibition in Ústí nad Labem in 1903.  Though this was not the first transmission of wireless telegraphy in Austria, as some newspapers claimed, it was probably the first transmission of wireless telegraphy over a large distance presented to the public in Bohemia. The idea of presenting wireless telegraphy at this exhibition was promoted by Wilhelm Biscan, director of the Elektrotechnikum school in Teplice at that time. The telegraph was installed by the Allgemeine Elektrizitäts-Gesellschaft in Berlin. The device used the Slaby-Arco system. 

  9. Non-invasive in-cell determination of free cytosolic [NAD+]/[NADH] ratios using hyperpolarized glucose show large variations in metabolic phenotypes

    DEFF Research Database (Denmark)

    Christensen, Caspar Elo; Karlsson, Magnus; Winther, Jakob R.

    2014-01-01

    Accumulating evidence suggest that the pyridine nucleotide NAD has far wider biological functions than its classical role in energy metabolism. NAD is used by hundreds of enzymes that catalyse substrate oxidation and as such it plays a key role in various biological processes such as aging, cell ......+]/[NADH] ratio, the bioprobe will enable better understanding of the origin of diverse pathological states of the cell as well as monitor cellular consequences of diseases and/or treatments.......Accumulating evidence suggest that the pyridine nucleotide NAD has far wider biological functions than its classical role in energy metabolism. NAD is used by hundreds of enzymes that catalyse substrate oxidation and as such it plays a key role in various biological processes such as aging, cell...... death and oxidative stress. It has been suggested that changes in the ratio of free cytosolic [NAD+]/[NADH] reflects metabolic alterations leading to, or correlating with, pathological states. We have designed an isotopically labelled metabolic bioprobe of free cytosolic [NAD+]/[NADH] by combining...

  10. A biochemical correlate of dimorphism in a zygomycete Benjaminiella poitrasii: characterization of purified NAD-dependent glutamate dehydrogenase, a target for antifungal agents.

    Science.gov (United States)

    Joshi, C V; Pathan, E K; Punekar, N S; Tupe, S G; Kapadnis, B P; Deshpande, M V

    2013-07-01

    The fungal organisms, especially pathogens, change their vegetative (Y, unicellular yeast and H, hypha) morphology reversibly for survival and proliferation in the host environment. NAD-dependent glutamate dehydrogenase (NAD-GDH, EC 1.4.1.2) from a non-pathogenic dimorphic zygomycete Benjaminiella poitrasii was previously reported to be an important biochemical correlate of the transition process. The enzyme was purified to homogeneity and characterized. It is a 371 kDa native molecular weight protein made up of four identical subunits. Kinetic studies showed that unlike other NAD-GDHs, it may act as an anabolic enzyme and has more affinity towards 2-oxoglutarate than L-glutamate. Chemical modifications revealed the involvement of single histidine and lysine residues in the catalytic activity of the enzyme. The phosphorylation and dephosphorylation study showed that the NAD-GDH is present in active phosphorylated form in hyphal cells of B. poitrasii. Two of the 1,2,3 triazole linked β-lactam-bile acid conjugates synthesized in the laboratory (B18, B20) were found to be potent inhibitors of purified NAD-GDH which also significantly affected Y-H transition in B. poitrasii. Furthermore, the compound B20 inhibited germ tube formation during Y-H transition in Candida albicans strains and Yarrowia lipolytica. The possible use of NAD-GDH as a target for antifungal agents is discussed.

  11. Intracellular NAMPT-NAD+-SIRT1 cascade improves post-ischaemic vascular repair by modulating Notch signalling in endothelial progenitors.

    Science.gov (United States)

    Wang, Pei; Du, Hui; Zhou, Can-Can; Song, Jie; Liu, Xingguang; Cao, Xuetao; Mehta, Jawahar L; Shi, Yi; Su, Ding-Feng; Miao, Chao-Yu

    2014-12-01

    Intracellular nicotinamide phosphoribosyltransferase (NAMPT) is the rate-limiting enzyme for nicotinamide adenine dinucleotide (NAD(+)) biosynthesis. This study investigated the role of NAMPT-mediated NAD(+) signalling in post-ischaemic vascular repair. Mouse hind-limb ischaemia up-regulated NAMPT expression and NAD(+) level in bone marrow (BM). Pharmacological inhibition of NAMPT by a chemical inhibitor FK866 impaired the mobilization of endothelial progenitor cells (EPCs) from BM upon ischaemic stress. Transgenic mice overexpressing NAMPT (Tg mice), but not H247A-mutant dominant-negative NAMPT (DN-Tg mice), exhibited enhanced capillary density, increased number of proliferating endothelial cells, improved blood flow recovery, and augmented collateral arterioles in the ischaemic limb. In cultured BM-derived EPCs, inhibition of NAMPT suppressed proliferation, migration, and tube formation, whereas overexpression of NAMPT induced opposite effects. The promoting effects of NAMPT on EPCs were abolished by silencing of sirtuin 1 (SIRT1), rather than silencing of SIRT2-7. Overexpression of NAMPT led to a SIRT1-depedent enhancement of Notch-1 intracellular domain deacetylation, which inhibited Delta-like ligand-4 (DLL4)-Notch signalling and thereby up-regulated of VEGFR-2 and VEGFR-3. Injection of recombinant VEGF induced a more pronounced EPC mobilization in Tg, but not in DN-Tg, mice. Furthermore, overexpression of NAMPT down-regulated Fringe family glycosyltransferases in a SIRT1-dependent manner, which rendered Notch more sensitive to the pro-angiogenic ligand Jagged1 rather than the anti-angiogenic ligand DLL4. These results demonstrate that intracellular NAMPT-NAD(+)-SIRT1 cascade improves post-ischaemic neovascularization. The modulation of Notch signalling may contribute to the enhanced post-ischaemic neovascularization. Published on behalf of the European Society of Cardiology. All rights reserved. © The Author 2014. For permissions please email: journals.permissions@oup.com.

  12. The mechanism of RNA 5' capping with NAD+, NADH and desphospho-CoA

    Energy Technology Data Exchange (ETDEWEB)

    Bird, Jeremy G.; Zhang, Yu; Tian, Yuan; Panova, Natalya; Barvík, Ivan; Greene, Landon; Liu, Min; Buckley, Brian; Krásný, Libor; Lee, Jeehiun K.; Kaplan, Craig D.; Ebright, Richard H.; Nickels, Bryce E.

    2016-07-06

    The chemical nature of the 5' end of RNA is a key determinant of RNA stability, processing, localization and translation efficiency and has been proposed to provide a layer of ‘epitranscriptomic’ gene regulation. Recently it has been shown that some bacterial RNA species carry a 5'-end structure reminiscent of the 5' 7-methylguanylate ‘cap’ in eukaryotic RNA. In particular, RNA species containing a 5'-end nicotinamide adenine dinucleotide (NAD+) or 3'-desphospho-coenzyme A (dpCoA) have been identified in both Gram-negative and Gram-positive bacteria. It has been proposed that NAD+, reduced NAD+ (NADH) and dpCoA caps are added to RNA after transcription initiation, in a manner analogous to the addition of 7-methylguanylate caps. Here we show instead that NAD+, NADH and dpCoA are incorporated into RNA during transcription initiation, by serving as non-canonical initiating nucleotides (NCINs) for de novo transcription initiation by cellular RNA polymerase (RNAP). We further show that both bacterial RNAP and eukaryotic RNAP II incorporate NCIN caps, that promoter DNA sequences at and upstream of the transcription start site determine the efficiency of NCIN capping, that NCIN capping occurs in vivo, and that NCIN capping has functional consequences. We report crystal structures of transcription initiation complexes containing NCIN-capped RNA products. Our results define the mechanism and structural basis of NCIN capping, and suggest that NCIN-mediated ‘ab initio capping’ may occur in all organisms.

  13. Structure of the Francisella tularensis enoyl-acyl carrier protein reductase (FabI) in complex with NAD+ and triclosan

    International Nuclear Information System (INIS)

    Mehboob, Shahila; Truong, Kent; Santarsiero, Bernard D.; Johnson, Michael E.

    2010-01-01

    Structure of the ternary complex of F. tularensis enoyl-acyl carrier protein reductase reveals the structure of the substrate binding loop whose electron density was missing in an earlier structure, and demonstrates a shift in the position of the NAD + cofactor. Enoyl-acyl carrier protein reductase (FabI) catalyzes the last rate-limiting step in the elongation cycle of the fatty-acid biosynthesis pathway and has been validated as a potential antimicrobial drug target in Francisella tularensis. The development of new antibiotic therapies is important both to combat potential drug-resistant bioweapons and to address the broader societal problem of increasing antibiotic resistance among many pathogenic bacteria. The crystal structure of FabI from F. tularensis (FtuFabI) in complex with the inhibitor triclosan and the cofactor NAD + has been solved to a resolution of 2.1 Å. Triclosan is known to effectively inhibit FabI from different organisms. Precise characterization of the mode of triclosan binding is required to develop highly specific inhibitors. Comparison of our structure with the previously determined FtuFabI structure which is bound to only NAD + reveals the conformation of the substrate-binding loop, electron density for which was missing in the earlier structure, and demonstrates a shift in the conformation of the NAD + cofactor. This shift in the position of the phosphate groups allows more room in the active site for substrate or inhibitor to bind and be better accommodated. This information will be crucial for virtual screening studies to identify novel scaffolds for development into new active inhibitors

  14. Environment dictates dependence on mitochondrial complex I for NAD+ and aspartate production and determines cancer cell sensitivity to metformin

    OpenAIRE

    Gui, Dan Y.; Sullivan, Lucas B.; Luengo, Alba; Hosios, Aaron M.; Bush, Lauren N.; Gitego, Nadege; Davidson, Shawn M.; Freinkman, Elizaveta; Thomas, Craig J.; Vander Heiden, Matthew G.

    2016-01-01

    Metformin use is associated with reduced cancer mortality, but how metformin impacts cancer outcomes is controversial. While metformin can act cell autonomously to inhibit tumor growth, the doses of metformin that inhibit proliferation in tissue culture are much higher than what has been described in vivo. Here, we show that environment drastically alters sensitivity to metformin and other complex I inhibitors. We find that complex I supports proliferation by regenerating NAD+, and metformin’...

  15. Neutral pion production measurements at SciBooNE

    International Nuclear Information System (INIS)

    Catala-Perez, J.

    2011-01-01

    Neutrino-induced neutral pion production is an important measurement for next generation neutrino oscillation experiments. Neutral current (NC) neutral pion production is a direct background for electron neutrino appearance experiments, while charged current (CC) neutral pion production affects experiments looking for muon neutrino disappearance. Located in the Booster Neutrino Beam at Fermilab, SciBooNE is a neutrino scattering experiment designed to accurately measure muon neutrino and anti-neutrino cross sections on carbon near 1 GeV neutrino energy. In this talk I will present recent SciBooNE results on neutral pion production, including the total cross section measurement for both channels relative to the CC inclusive cross section, the separation of the coherent and incoherent contributions to the NC channel, and details on neutral pion production kinematics.

  16. MicroBooNE and its Cross Section Measurement

    Energy Technology Data Exchange (ETDEWEB)

    Tsai, Yun-Tse [SLAC

    2017-05-22

    MicroBooNE (the Micro Booster Neutrino Experiment) is a short-baseline neutrino experiment based on the technology of a liquid-argon time-projection chamber (LArTPC), and has recently completed its first year of data-taking in the Fermilab Booster Neutrino Beam. It aims to address the anomalous excess of events with an electromagnetic final state in MiniBooNE, to measure neutrino-argon interaction cross sections, and to provide relevant R\\&D for the future LArTPC experiments, such as DUNE. In these proceedings, we present the first reconstructed energy spectrum of Michel electrons from cosmic muon decays, the first kinematic distributions of the candidate muon tracks from $\

  17. A Low-Li Geochemical Province in the NE Atlantic

    DEFF Research Database (Denmark)

    Bailey, J. C.; Gwozdz, R.

    1978-01-01

    Lithium was analysed in 392 basalts and related igneous rocks from the North Atlantic Tertiary-Recent province using activation analysis and Čerenkov counting. Monotonous Li values of 5.5±2 ppm in NE Atlantic basalts define a low-Li geochemical province which has persisted for 60 million years......, over 20° of latitude and regardless of basalt type and chemistry. This low-Li province and the increasing Li contents of ocean-ridge tholeiites into the S Atlantic are believed to monitor Li heterogeneity in the underlying mantle. Li, like Na, increases gently during the differentiation of several...... basalt series. No whole-rock coherence is observed between Li and Mg, K, Rb or Ca. Mantle phlogopite is considered to play an insignificant rôle in controlling the Li levels of NE Atlantic basalts....

  18. Replicatively senescent human fibroblasts reveal a distinct intracellular metabolic profile with alterations in NAD+ and nicotinamide metabolism.

    Science.gov (United States)

    James, Emma L; Lane, James A E; Michalek, Ryan D; Karoly, Edward D; Parkinson, E Kenneth

    2016-12-07

    Cellular senescence occurs by proliferative exhaustion (PEsen) or following multiple cellular stresses but had not previously been subject to detailed metabolomic analysis. Therefore, we compared PEsen fibroblasts with proliferating and transiently growth arrested controls using a combination of different mass spectroscopy techniques. PEsen cells showed many specific alterations in both the NAD+ de novo and salvage pathways including striking accumulations of nicotinamide mononucleotide (NMN) and nicotinamide riboside (NR) in the amidated salvage pathway despite no increase in nicotinamide phosphoribosyl transferase or in the NR transport protein, CD73. Extracellular nicotinate was depleted and metabolites of the deamidated salvage pathway were reduced but intracellular NAD+ and nicotinamide were nevertheless maintained. However, sirtuin 1 was downregulated and so the accumulation of NMN and NR was best explained by reduced flux through the amidated arm of the NAD+ salvage pathway due to reduced sirtuin activity. PEsen cells also showed evidence of increased redox homeostasis and upregulated pathways used to generate energy and cellular membranes; these included nucleotide catabolism, membrane lipid breakdown and increased creatine metabolism. Thus PEsen cells upregulate several different pathways to sustain their survival which may serve as pharmacological targets for the elimination of senescent cells in age-related disease.

  19. The cAMP/PKA pathway rapidly activates SIRT1 to promote fatty acid oxidation independently of changes in NAD(+).

    Science.gov (United States)

    Gerhart-Hines, Zachary; Dominy, John E; Blättler, Sharon M; Jedrychowski, Mark P; Banks, Alexander S; Lim, Ji-Hong; Chim, Helen; Gygi, Steven P; Puigserver, Pere

    2011-12-23

    The NAD(+)-dependent deacetylase SIRT1 is an evolutionarily conserved metabolic sensor of the Sirtuin family that mediates homeostatic responses to certain physiological stresses such as nutrient restriction. Previous reports have implicated fluctuations in intracellular NAD(+) concentrations as the principal regulator of SIRT1 activity. However, here we have identified a cAMP-induced phosphorylation of a highly conserved serine (S434) located in the SIRT1 catalytic domain that rapidly enhanced intrinsic deacetylase activity independently of changes in NAD(+) levels. Attenuation of SIRT1 expression or the use of a nonphosphorylatable SIRT1 mutant prevented cAMP-mediated stimulation of fatty acid oxidation and gene expression linked to this pathway. Overexpression of SIRT1 in mice significantly potentiated the increases in fatty acid oxidation and energy expenditure caused by either pharmacological β-adrenergic agonism or cold exposure. These studies support a mechanism of Sirtuin enzymatic control through the cAMP/PKA pathway with important implications for stress responses and maintenance of energy homeostasis. Copyright © 2011 Elsevier Inc. All rights reserved.

  20. UNIVERSAL JURISDICTION AND THE PRINCIPLE OF NE BIS IN IDEM

    Directory of Open Access Journals (Sweden)

    MIHAELA AGHENITEI

    2011-04-01

    Full Text Available Universal jurisdiction was defined as “the assertion of jurisdiction to prescribe in the absence of any other accepted jurisdictional nexus at the time of the relevant conduct.” Professor Randall, in his seminal work on universal jurisdiction, opined that the theory of universality “provides every state with jurisdiction over a limitedcategory of offenses generally recognized as of universal concern, regardless of the situs of the offence and the nationalities of the offender and the offended. ”Universal jurisdiction is considered a tool for promoting greater justice, but the rights of the accused must be protected. One of the most important guarantees is the principle of ne bis in idem, which protected persons against multiple prosecutions for the same crime. The main legal consequence of the application of ne bis in idem in most systems is the prohibition and inadmissibility of subsequent prosecutions on the same facts blocking effect.The national ne bis in idem principle is established asan individual right in international human rights legal instruments, such as the International Covenant on Civil and Political Rights of 19 December 1966, in Article 14(7. At the regional level, Article 8(4 of the American Convention of Human Rights (1969 and Article 4 (I of the Seventh Protocol of the European Convention of Human Rights merit mention. In Europe, the ne bis in idem principle is enshrined in Article 54 of the Convention implementing the Schengen Agreement of 14 June 1985, which prohibits the initiation of a second trial for the same offence when final judgment has been imposed upon a person by a court of a contracting party.

  1. The KM3NeT Digital Optical Module

    Directory of Open Access Journals (Sweden)

    Vivolo Daniele

    2016-01-01

    Full Text Available KM3NeT is a European deep-sea multidisciplinary research infrastructure in the Mediterranean Sea. It will host a km3-scale neutrino telescope and dedicated instruments for long-term and continuous measurements for Earth and Sea sciences. The KM3NeT neutrino telescope is a 3-dimensional array of Digital Optical Modules, suspended in the sea by means of vertical string structures, called Detection Units, supported by two pre-stretched Dyneema ropes, anchored to the seabed and kept taut with a system of buoys. The Digital Optical Module represents the active part of the neutrino telescope. It is composed by a 17-inch, 14 mm thick borosilicate glass (Vitrovex spheric vessel housing 31 photomultiplier tubes with 3-inch photocathode diameter and the associated front-end and readout electronics. The technical solution adopted for the KM3NeT optical modules is characterized by an innovative design, considering that existing neutrino telescopes, Baikal, IceCube and ANTARES, all use large photomultipliers, typically with a diameter of 8″ or 10″. It offers several advantages: higher sensitive surface (1260 cm2, weaker sensitivity to Earth's magnetic field, better distinction between single-photon and multi-photon events (photon counting and directional information with an almost isotropic field of view. In this contribution the design and the performance of the KM3NeT Digital Optical Modules are discussed, with a particular focus on enabling technologies and integration procedure.

  2. Calibration of a NE213 detector for neutron spectroscopy

    International Nuclear Information System (INIS)

    Blazquez Martinez, J.; Butragueno Casado, J. L.

    1974-01-01

    This work describes the experimental way followed for getting the calibration of a NE213 detector with a beam of neutrons from the J.E.N. 2 MeV Van de Graaff and using at once pulse shape discrimination. Detector has been used for measuring the spectrum of the fast reactor CORAL-1. There is also included an experimental method in order to get with precision where the Compton edge is placed on the electron spectrum. (Author) 9 refs

  3. U,Th-21Ne dating and its applications

    International Nuclear Information System (INIS)

    Basu, Sudeshna; Murty, S.V.S.; Anil Kumar

    2003-01-01

    The potential of radiogenic and fissiogenic noble gas isotopes as dating tools has been well exploited. U, Th- 4 He , K- 40 Ar and U- fission Xe pairs as well as their variants like 39 Ar- 40 Ar and induced fission Xe- spontaneous fission Xe pairs have been extensively used as geochronological tools. A new dating method that utilizes the nucleogenic isotope 21 Ne and demonstrate its application for an apatite separate from a carbonatite is proposed

  4. Modeling Ne-21 NMR parameters for carbon nanosystems

    Czech Academy of Sciences Publication Activity Database

    Kupka, T.; Nieradka, M.; Kaminský, Jakub; Stobinski, L.

    2013-01-01

    Roč. 51, č. 10 (2013), s. 676-681 ISSN 0749-1581 R&D Projects: GA ČR GAP208/11/0105; GA MŠk(CZ) LH11033 Grant - others:AV ČR(CZ) M200551205 Institutional support: RVO:61388963 Keywords : Ne-21 NMR * GIAO NMR * molecular modeling * carbon nanostructures Subject RIV: CF - Physical ; Theoretical Chemistry Impact factor: 1.559, year: 2013

  5. Monte Carlo studies for the KM3NeT neutrino telescope

    Energy Technology Data Exchange (ETDEWEB)

    Shanidze, Rezo [ECAP, Universitaet Erlangen-Nuernberg (Germany)

    2010-07-01

    KM3NeT is a future European deep-sea research infrastructure in the Mediterranean Sea, which will host a high energy neutrino telescope with a multi-km{sup 3} instrumented volume. Recently, the KM3NeT consortium, which is formed by the ANTARES, NEMO and NESTOR collaborations as well as marine science and technology institutes, released the KM3NeT technical design report (TDR) document. The KM3NeT design options presented in the KM3NeT TDR and the results of corresponding Monte Carlo studies are discussed in the talk.

  6. Nad knižkou Genius loci - (rozhovory o Slovensku: malý pokus o autorecenziu

    Directory of Open Access Journals (Sweden)

    Mikuláš Huba

    2011-12-01

    Full Text Available Ak spätne rozmýšľam nad tým, prečo som prijal ponuku Krás Slovenska robiť rozhovory so známymi ľuďmi s blízkym vzťahom k Slovensku a jeho ohrozeným hodnotám, a to napriek tomu, že dosiaľ som sa nezvykol pýtať ja, ale skôr sa pýtali iní mňa (najskôr učitelia, potom novinári a občas aj súdruhovia zo Štátnej bezpečnosti, zisťujem, že dôvody môjho odvážneho súhlasu a zároveň kritériá výberu „respondentov“ boli najmä tieto:

  7. Regulation of Serine-Threonine Kinase Akt Activation by NAD+-Dependent Deacetylase SIRT7

    Directory of Open Access Journals (Sweden)

    Jia Yu

    2017-01-01

    Full Text Available The Akt pathway is a central regulator that promotes cell survival in response to extracellular signals. Depletion of SIRT7, an NAD+-dependent deacetylase that is the least-studied sirtuin, is known to significantly increase Akt activity in mice through unknown mechanisms. In this study, we demonstrate that SIRT7 depletion in breast cancer cells results in Akt hyper-phosphorylation and increases cell survival following genotoxic stress. Mechanistically, SIRT7 specifically interacts with and deacetylates FKBP51 at residue lysines 28 and 155 (K28 and K155, resulting in enhanced interactions among FKBP51, Akt, and PHLPP, as well as Akt dephosphorylation. Mutating both lysines to arginines abolishes the effect of SIRT7 on Akt activity through FKBP51 deacetylation. Finally, energy stress strengthens SIRT7-mediated effects on Akt dephosphorylation through FKBP51 and thus sensitizes cancer cells to cytotoxic agents. These results reveal a direct role of SIRT7 in Akt regulation and raise the possibility of using the glucose analog 2-deoxy-D-glucose (2DG as a chemo-sensitizing agent.

  8. Subattomole detection of adiponectin in urine by ultrasensitive ELISA coupled with thio-NAD cycling

    Science.gov (United States)

    Morikawa, Mika; Naito, Rina; Mita, Koichi; Watabe, Satoshi; Nakaishi, Kazunari; Yoshimura, Teruki; Miura, Toshiaki; Hashida, Seiichi; Ito, Etsuro

    2015-01-01

    Adiponectin is a hormone secreted from adipocytes, and it demonstrates antidiabetic, anti-atherosclerotic, antiobesity and anti-inflammatory effects. However, the patterns of change in urinary adiponectin levels in various diseases remain unknown, because only trace amounts of the hormone are present in urine. In the present study, we applied an ultrasensitive ELISA coupled with thio-NAD cycling to measure urinary adiponectin levels. Spikeand-recovery tests using urine confirmed the reliability of our ultrasensitive ELISA. The limit of detection for adiponectin in urine was 2.3×10−19 moles/assay (1.4 pg/mL). The urinary adiponectin concentration ranged between 0.04 and 5.82 ng/mL in healthy subjects. The pilot study showed that the urinary adiponectin levels, which were corrected by the creatinine concentration, were 0.73±0.50 (ng/mg creatinine, N=6) for healthy subjects, versus 12.02±3.85 (ng/mg creatinine, N=3) for patients with diabetes mellitus (DM). That is, the urinary adiponectin levels were higher (P<0.05) in DM patients than in healthy subjects. Further, these urinary adiponectin levels tended to increase with the progression of DM accompanied with nephropathy. Our method is thus expected to provide a simple, rapid and reasonably priced test for noninvasive monitoring of the progression of DM without the requirement of special tools. PMID:27493857

  9. Allele polymorphism of Nad1 gene of the Serbian spruce mitochondrial genome

    Directory of Open Access Journals (Sweden)

    Milovanović Jelena

    2007-01-01

    Full Text Available Serbian spruce (Picea omorika /Panč./Purkyne, as the Balkan Peninsula endemic and the Tertiary relic, is a species whose survival is threatened by the constant restriction of its range caused by the global changes of environmental conditions and the adverse human impacts. The Serbian spruce seedling seed orchard at Godovik represents the base for the improvement of the production of the selected seeds of this species, which can be used as the initial material for the extension of its range. The allele polymorphism of the mitochondrial nad1 gene was analyzed in five different Serbian spruce phenogroups of which the orchard is established. The obtained results are a contribution to a closer study of the causes of the postglacial intraspecific differentiation of Serbian spruce and the creation of the above phenogroups. The study results are significant for further breeding of this species based on the better knowledge of the genetic structure of the species, its directed utilisation and the widening of its range. .

  10. Beta-lapachone, a modulator of NAD metabolism, prevents health declines in aged mice.

    Directory of Open Access Journals (Sweden)

    Jeong-sook Lee

    Full Text Available NADH-quinone oxidoreductase 1 (NQO1 modulates cellular NAD(+/NADH ratio which has been associated with the aging and anti-aging mechanisms of calorie restriction (CR. Here, we demonstrate that the facilitation of NQO1 activity by feeding β-lapachone (βL, an exogenous NQO1 co-substrate, prevented age-dependent decline of motor and cognitive function in aged mice. βL-fed mice did not alter their food-intake or locomotor activity but did increase their energy expenditure as measured by oxygen consumption and heat generation. Mitochondrial structure and numbers were disorganized and decreased in the muscles of control diet group but those defects were less severe in βL-fed aged mice. Furthermore, for a subset of genes associated with energy metabolism, mice fed the βL-diet showed similar changes in gene expression to the CR group (fed 70% of the control diet. These results support the potentiation of NQO1 activity by a βL diet and could be an option for preventing age-related decline of muscle and brain functions.

  11. NAD+-dependent HDAC inhibitor stimulates Monascus pigment production but inhibit citrinin.

    Science.gov (United States)

    Hu, Yan; Zhou, Youxiang; Mao, Zejing; Li, Huihui; Chen, Fusheng; Shao, Yanchun

    2017-08-23

    Monascus species are edible fungi due to the production of food colorant and other beneficial compounds. Hence, it has been an attractive thesis to improve their productivities. Increasing numbers of investigations revealed that regulating the activities of histone deacetylases can significantly perturb secondary metabolites (SM) production at a global level. In this study, dihydrocoumarin (DHC, an inhibitor of the Sirtuin family of NAD + -dependent deacetylases) was used to treat Monascus ruber for evaluating its effects on organism growth and SM production. The results revealed that the variation trends of colonial sizes, biomass and mycotoxin were in a dose-dependent manner. Generally, they decreased with the increased DHC concentrations in the designed range. But the variation trend of pigment was different. Comparison of SM profile, three new peaks occurred to the mycelia extractions from DHC-treated strain corresponding to molecular weights 402, 416 and 444, respectively. These three compounds were identified as Monasfluol B, Monascus azaphilone C and acetyl-monasfluol B (a new Monascus chemical pigment structure). In short, DHC can stimulate M. ruber strain to produce more pigment-like polyketides but inhibition of mycotoxin (citrinin).

  12. Visual transactions and reinscriptions of identity in Nadín Ospina and Calimocho Styles

    Directory of Open Access Journals (Sweden)

    María Elena Lucero

    2013-07-01

    Full Text Available Relecting on identities in Latin America in the space of visual arts leads to an analytical revision not only of alternative versions of the cultural past, but also of the way in which transnationality has affected contemporary practices, especially through extended periods of exile, diaspora, migrations and displacements. One possible conceptual weapon would be the use of irony or parody. In this aspect, the Colombian artist Nadín Ospina synthesizes possible exchanges between symbolic productions of the Pre-Hispanic period, current fetishized merchandise, and certain imaginaries linked to the media. Projected from the place of sarcasm, Ospina materializes these fetish objects that allude to the exotic nickna megiven by Europe to American visuality andresets cultural difference to the extreme of potentializingit and putting it in lux. On the other hand, Calimocho Styles, the duo between the Mexicanartists Ruben Ortiz Torres and Eduardo Abaroa examines unstable and changing cultural identities that emerge from the continuous border crossing between Mexico and United States, inquiring into ambivalent strategies of generating cultural products and proposing contemporary ictions from iconic referents of mass consumption. In both cases, hybrid visual proposals are generated, which create a space for numerous inquiries and discussions on interculturality and its incidence in Latin America.

  13. An Interbacterial NAD(P)+ Glycohydrolase Toxin Requires Elongation Factor Tu for Delivery to Target Cells

    Energy Technology Data Exchange (ETDEWEB)

    Whitney, John C.; Quentin, Dennis; Sawai, Shin; LeRoux, Michele; Harding, Brittany N.; Ledvina, Hannah E.; Tran, Bao Q.; Robinson, Howard; Goo, Young Ah; Goodlett, David R.; Raunser, Stefan; Mougous, Joseph D.

    2015-10-08

    Type VI secretion (T6S) influences the composition of microbial communities by catalyzing the delivery of toxins between adjacent bacterial cells. Here, we demonstrate that a T6S integral membrane toxin from Pseudomonas aeruginosa, Tse6, acts on target cells by degrading the universally essential dinucleotides NAD+ and NADP+. Structural analyses of Tse6 show that it resembles mono-ADP-ribosyltransferase proteins, such as diphtheria toxin, with the exception of a unique loop that both excludes proteinaceous ADP-ribose acceptors and contributes to hydrolysis. We find that entry of Tse6 into target cells requires its binding to an essential housekeeping protein, translation elongation factor Tu (EF-Tu). These proteins participate in a larger assembly that additionally directs toxin export and provides chaperone activity. Visualization of this complex by electron microscopy defines the architecture of a toxin-loaded T6S apparatus and provides mechanistic insight into intercellular membrane protein delivery between bacteria.

  14. Human mitochondrial NAD(P)(+)-dependent malic enzyme participates in cutaneous melanoma progression and invasion.

    Science.gov (United States)

    Chang, Yung-Lung; Gao, Hong-Wei; Chiang, Chien-Ping; Wang, Wei-Ming; Huang, Shih-Ming; Ku, Chien-Fen; Liu, Guang-Yaw; Hung, Hui-Chih

    2015-03-01

    Cutaneous melanoma is the most life-threatening neoplasm of the skin, accounting for most of the skin cancer deaths. Accumulating evidence suggests that targeting metabolism is an appealing strategy for melanoma therapy. Mitochondrial NAD(P)(+)-dependent malic enzyme (ME2), an oxidative decarboxylase, was evaluated for its biological significance in cutaneous melanoma progression. ME2 mRNA and protein expression significantly increased during melanoma progression, as evidenced by Gene Expression Omnibus analysis and immunohistochemistry on clinically annotated tissue microarrays, respectively. In addition, ME2 knockdown attenuated melanoma cell proliferation in vitro. ME2 ablation resulted in reduced cellular ATP levels and elevated cellular reactive oxygen species production, which activated the AMP-activated protein kinase pathway and inhibited acetyl-CoA carboxylase. Furthermore, ME2 expression was associated with cell migration and invasion. ME2 knockdown decreased anchorage-independent growth in vitro and tumor cell growth in vivo. These results suggested that ME2 might be an important factor in melanoma progression and a novel biomarker of invasion.

  15. Estimation of the effective population size (Ne) and its application in the management of small populations

    DEFF Research Database (Denmark)

    Jimenez Mena, Belen

    2016-01-01

    Effective population size (Ne) is an important concept to understand the evolution of a population. In conservation, Ne is used to assess the threat status of a population, evaluate its genetic viability in the future and set conservation priorities. An accurate estimation of Ne is thus essential....... The main objective of this thesis was to better understand how the estimation of Ne using molecular markers can be improved for use in conservation genetics. As a first step, we undertook a simulation study where three different methods to estimate Ne were investigated. We explored how well these three...... methods performed under different scenarios. This study showed that all three methods performed better when the number of unlinked loci used to make the estimation increased and the minimum number of loci need for an accurate estimation of Ne was 100 SNPs. A general assumption in the estimation of Ne...

  16. The gdhB gene of Pseudomonas aeruginosa encodes an arginine-inducible NAD(+)-dependent glutamate dehydrogenase which is subject to allosteric regulation.

    Science.gov (United States)

    Lu, C D; Abdelal, A T

    2001-01-01

    The NAD(+)-dependent glutamate dehydrogenase (NAD-GDH) from Pseudomonas aeruginosa PAO1 was purified, and its amino-terminal amino acid sequence was determined. This sequence information was used in identifying and cloning the encoding gdhB gene and its flanking regions. The molecular mass predicted from the derived sequence for the encoded NAD-GDH was 182.6 kDa, in close agreement with that determined from sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the purified enzyme (180 kDa). Cross-linking studies established that the native NAD-GDH is a tetramer of equal subunits. Comparison of the derived amino acid sequence of NAD-GDH from P. aeruginosa with the GenBank database showed the highest homology with hypothetical polypeptides from Pseudomonas putida, Mycobacterium tuberculosis, Rickettsia prowazakii, Legionella pneumophila, Vibrio cholerae, Shewanella putrefaciens, Sinorhizobium meliloti, and Caulobacter crescentus. A moderate degree of homology, primarily in the central domain, was observed with the smaller tetrameric NAD-GDH (protomeric mass of 110 kDa) from Saccharomyces cerevisiae or Neurospora crassa. Comparison with the yet smaller hexameric GDH (protomeric mass of 48 to 55 kDa) of other prokaryotes yielded a low degree of homology that was limited to residues important for binding of substrates and for catalytic function. NAD-GDH was induced 27-fold by exogenous arginine and only 3-fold by exogenous glutamate. Primer extension experiments established that transcription of gdhB is initiated from an arginine-inducible promoter and that this induction is dependent on the arginine regulatory protein, ArgR, a member of the AraC/XyIS family of regulatory proteins. NAD-GDH was purified to homogeneity from a recombinant strain of P. aeruginosa and characterized. The glutamate saturation curve was sigmoid, indicating positive cooperativity in the binding of glutamate. NAD-GDH activity was subject to allosteric control by arginine and citrate, which

  17. Dual roles of Lys(57) at the dimer interface of human mitochondrial NAD(P)+-dependent malic enzyme.

    Science.gov (United States)

    Hsieh, Ju-Yi; Liu, Jyung-Hurng; Fang, Yi-Wen; Hung, Hui-Chih

    2009-05-13

    Human m-NAD(P)-ME [mitochondrial NAD(P)+-dependent ME (malic enzyme)] is a homotetramer, which is allosterically activated by the binding of fumarate. The fumarate-binding site is located at the dimer interface of the NAD(P)-ME. In the present study, we decipher the functional role of the residue Lys57, which resides at the fumarate-binding site and dimer interface, and thus may be involved in the allosteric regulation and subunit-subunit interaction of the enzyme. In the present study, Lys57 is replaced with alanine, cysteine, serine and arginine residues. Site-directed mutagenesis and kinetic analysis strongly suggest that Lys57 is important for the fumarate-induced activation and quaternary structural organization of the enzyme. Lys57 mutant enzymes demonstrate a reduction of Km and an elevation of kcat following induction by fumarate binding, and also display a much higher maximal activation threshold than WT (wild-type), indicating that these Lys57 mutant enzymes have lower affinity for the effector fumarate. Furthermore, mutation of Lys57 in m-NAD(P)-ME causes the enzyme to become less active and lose co-operativity. It also increased K0.5,malate and decreased kcat values, indicating that the catalytic power of these mutant enzymes was significantly impaired following mutation of Lys57. Analytical ultracentrifugation analysis demonstrates that the K57A, K57S and K57C mutant enzymes dissociate predominantly into dimers, with some monomers present, whereas the K57R mutant forms a mixture of dimers and tetramers, with a small amount of the enzyme in monomeric form. The dimeric form of these Lys57 mutants, however, cannot be reconstituted into tetramers with the addition of fumarate. Modelling structures of the Lys57 mutant enzymes show that the hydrogen bond network in the dimer interface where Lys57 resides may be reduced compared with WT. Although the fumarate-induced activation effects are partially maintained in these Lys57 mutant enzymes, the mutant enzymes

  18. Calibration of a NE213 detector for neutron spectroscopy; Calibracion de un detector de NE213 para espectroscopia de neutrones

    Energy Technology Data Exchange (ETDEWEB)

    Blazquez Martinez, J.; Butragueno Casado, J. L.

    1974-07-01

    This work describes the experimental way followed for getting the calibration of a NE213 detector with a beam of neutrons from the J.E.N. 2 MeV Van de Graaff and using at once pulse shape discrimination. Detector has been used for measuring the spectrum of the fast reactor CORAL-1. There is also included an experimental method in order to get with precision where the Compton edge is placed on the electron spectrum. (Author) 9 refs.

  19. NAD+ Levels Control Ca2+ Store Replenishment and Mitogen-induced Increase of Cytosolic Ca2+ by Cyclic ADP-ribose-dependent TRPM2 Channel Gating in Human T Lymphocytes*

    Science.gov (United States)

    Magnone, Mirko; Bauer, Inga; Poggi, Alessandro; Mannino, Elena; Sturla, Laura; Brini, Marisa; Zocchi, Elena; De Flora, Antonio; Nencioni, Alessio; Bruzzone, Santina

    2012-01-01

    Intracellular NAD+ levels ([NAD+]i) are important in regulating human T lymphocyte survival, cytokine secretion, and the capacity to respond to antigenic stimuli. NAD+-derived Ca2+-mobilizing second messengers, produced by CD38, play a pivotal role in T cell activation. Here we demonstrate that [NAD+]i modifications in T lymphocytes affect intracellular Ca2+ homeostasis both in terms of mitogen-induced [Ca2+]i increase and of endoplasmic reticulum Ca2+ store replenishment. Lowering [NAD+]i by FK866-mediated nicotinamide phosphoribosyltransferase inhibition decreased the mitogen-induced [Ca2+]i rise in Jurkat cells and in activated T lymphocytes. Accordingly, the Ca2+ content of thapsigargin-sensitive Ca2+ stores was greatly reduced in these cells in the presence of FK866. When NAD+ levels were increased by supplementing peripheral blood lymphocytes with the NAD+ precursors nicotinamide, nicotinic acid, or nicotinamide mononucleotide, the Ca2+ content of thapsigargin-sensitive Ca2+ stores as well as cell responsiveness to mitogens in terms of [Ca2+]i elevation were up-regulated. The use of specific siRNA showed that the changes of Ca2+ homeostasis induced by NAD+ precursors are mediated by CD38 and the consequent ADPR-mediated TRPM2 gating. Finally, the presence of NAD+ precursors up-regulated important T cell functions, such as proliferation and IL-2 release in response to mitogens. PMID:22547068

  20. NAD+ levels control Ca2+ store replenishment and mitogen-induced increase of cytosolic Ca2+ by Cyclic ADP-ribose-dependent TRPM2 channel gating in human T lymphocytes.

    Science.gov (United States)

    Magnone, Mirko; Bauer, Inga; Poggi, Alessandro; Mannino, Elena; Sturla, Laura; Brini, Marisa; Zocchi, Elena; De Flora, Antonio; Nencioni, Alessio; Bruzzone, Santina

    2012-06-15

    Intracellular NAD(+) levels ([NAD(+)](i)) are important in regulating human T lymphocyte survival, cytokine secretion, and the capacity to respond to antigenic stimuli. NAD(+)-derived Ca(2+)-mobilizing second messengers, produced by CD38, play a pivotal role in T cell activation. Here we demonstrate that [NAD(+)](i) modifications in T lymphocytes affect intracellular Ca(2+) homeostasis both in terms of mitogen-induced [Ca(2+)](i) increase and of endoplasmic reticulum Ca(2+) store replenishment. Lowering [NAD(+)](i) by FK866-mediated nicotinamide phosphoribosyltransferase inhibition decreased the mitogen-induced [Ca(2+)](i) rise in Jurkat cells and in activated T lymphocytes. Accordingly, the Ca(2+) content of thapsigargin-sensitive Ca(2+) stores was greatly reduced in these cells in the presence of FK866. When NAD(+) levels were increased by supplementing peripheral blood lymphocytes with the NAD(+) precursors nicotinamide, nicotinic acid, or nicotinamide mononucleotide, the Ca(2+) content of thapsigargin-sensitive Ca(2+) stores as well as cell responsiveness to mitogens in terms of [Ca(2+)](i) elevation were up-regulated. The use of specific siRNA showed that the changes of Ca(2+) homeostasis induced by NAD(+) precursors are mediated by CD38 and the consequent ADPR-mediated TRPM2 gating. Finally, the presence of NAD(+) precursors up-regulated important T cell functions, such as proliferation and IL-2 release in response to mitogens.

  1. How does a Poem Think? Czesław Miłosz Nad strumieniem

    Directory of Open Access Journals (Sweden)

    Joanna Zach

    2014-01-01

    Full Text Available The subject of the analysis is the poem by Czesław Miłosz from the volume To, entitled “Nad strumieniem”. The task the author embarked upon boils down to the answer to the question: how does a specific idea(its hidden mental message develop in this poem and what results from it for the idea as well as for the poem. The proposed reading takes place at three levels. Preliminary reconnaissance is made by naive reading consisting in the reconstruction of the situation portrayed in the poem, in the recreation of that which can be fund, so to say, on the “surface” of the poem. Next, the author launches these contexts which in Miłosz’s representation of the attitude of man towards nature allow one to notice balance and harmony, coming to terms with life and wisdom, which is based on distance and silence. At the third reading level, the author refers to the image crowning opus magnum by Charles Darwin, in which a similar situation—contemplation in the open nature—leads to the formulation of the answer to the question about the sense of “fight in nature, famine and death”. This answer is juxtaposed with the ending of Czesław Miłosz’s poem: „Wydaje mi się, że słyszę głos demiurga:/ Albo nieme skały jak w pierwszym dniu stworzenia,/ albo życie, którego warunkiem jest śmierć,/ i to upajające ciebie piękno”. („It seems to me that I hear the voice of demiurge:/ Either silent rocks as on the first day of creation, / or life, whose condition is death,/ and this beauty engulfing you”.

  2. Update of the NAD(PH:quinone oxidoreductase (NQO gene family

    Directory of Open Access Journals (Sweden)

    Vasiliou Vasilis

    2006-03-01

    Full Text Available Abstract The NAD(PH:quinone acceptor oxidoreductase (NQO gene family belongs to the flavoprotein clan and, in the human genome, consists of two genes (NQO1 and NQO2. These two genes encode cytosolic flavoenzymes that catalyse the beneficial two-electron reduction of quinones to hydroquinones. This reaction prevents the unwanted one-electron reduction of quinones by other quinone reductases; one-electron reduction results in the formation of reactive oxygen species, generated by redox cycling of semiquinones in the presence of molecular oxygen. Both the mammalian NQO1 and NQO2 genes are upregulated as a part of the oxidative stress response and are inexplicably overexpressed in particular types of tumours. A non-synonymous mutation in the NQO1 gene, leading to absence of enzyme activity, has been associated with an increased risk of myeloid leukaemia and other types of blood dyscrasia in workers exposed to benzene. NQO2 has a melatonin-binding site, which may explain the anti-oxidant role of melatonin. An ancient NQO3 subfamily exists in eubacteria and the authors suggest that there should be additional divisions of the NQO family to include the NQO4 subfamily in fungi and NQO5 subfamily in archaebacteria. Interestingly, no NQO genes could be identified in the worm, fly, sea squirt or plants; because these taxa carry quinone reductases capable of one- and two-electron reductions, there has been either convergent evolution or redundancy to account for the appearance of these enzyme functions whenever they have been needed during evolution.

  3. M. tuberculosis Sliding β-Clamp Does Not Interact Directly with the NAD+ -Dependent DNA Ligase

    Science.gov (United States)

    Kukshal, Vandna; Khanam, Taran; Chopra, Deepti; Singh, Nidhi; Sanyal, Sabyasachi; Ramachandran, Ravishankar

    2012-01-01

    The sliding β-clamp, an important component of the DNA replication and repair machinery, is drawing increasing attention as a therapeutic target. We report the crystal structure of the M. tuberculosis β-clamp (Mtbβ-clamp) to 3.0 Å resolution. The protein crystallized in the space group C2221 with cell-dimensions a = 72.7, b = 234.9 & c = 125.1 Å respectively. Mtbβ-clamp is a dimer, and exhibits head-to-tail association similar to other bacterial clamps. Each monomer folds into three domains with similar structures respectively and associates with its dimeric partner through 6 salt-bridges and about 21 polar interactions. Affinity experiments involving a blunt DNA duplex, primed-DNA and nicked DNA respectively show that Mtbβ-clamp binds specifically to primed DNA about 1.8 times stronger compared to the other two substrates and with an apparent Kd of 300 nM. In bacteria like E. coli, the β-clamp is known to interact with subunits of the clamp loader, NAD+ -dependent DNA ligase (LigA) and other partners. We tested the interactions of the Mtbβ-clamp with MtbLigA and the γ-clamp loader subunit through radioactive gel shift assays, size exclusion chromatography, yeast-two hybrid experiments and also functionally. Intriguingly while Mtbβ-clamp interacts in vitro with the γ-clamp loader, it does not interact with MtbLigA unlike in bacteria like E. coli where it does. Modeling studies involving earlier peptide complexes reveal that the peptide-binding site is largely conserved despite lower sequence identity between bacterial clamps. Overall the results suggest that other as-yet-unidentified factors may mediate interactions between the clamp, LigA and DNA in mycobacteria. PMID:22545130

  4. Molecular evolutionary patterns of NAD+/Sirtuin aging signaling pathway across taxa.

    Directory of Open Access Journals (Sweden)

    Uma Gaur

    Full Text Available A deeper understanding of the conserved molecular mechanisms in different taxa have been made possible only because of the evolutionary conservation of crucial signaling pathways. In the present study, we explored the molecular evolutionary pattern of selection signatures in 51 species for 10 genes which are important components of NAD+/Sirtuin pathway and have already been directly linked to lifespan extension in worms and mice. Selection pressure analysis using PAML program revealed that MRPS5 and PPARGC1A were under significant constraints because of their functional significance. FOXO3a also displayed strong purifying selection. All three sirtuins, which were SIRT1, SIRT2 and SIRT6, displayed a great degree of conservation between taxa, which is consistent with the previous report. A significant evolutionary constraint is seen on the anti-oxidant gene, SOD3. As expected, TP53 gene was under significant selection pressure in mammals, owing to its major role in tumor progression. Poly-ADP-ribose polymerase (PARP genes displayed the most sites under positive selection. Further 3D structural analysis of PARP1 and PARP2 protein revealed that some of these positively selected sites caused a change in the electrostatic potential of the protein structure, which may allow a change in its interaction with other proteins and molecules ultimately leading to difference in the function. Although the functional significance of the positively selected sites could not be established in the variants databases, yet it will be interesting to see if these sites actually affect the function of PARP1 and PARP2.

  5. A transportable methane stabilized He-Ne laser

    Science.gov (United States)

    Akimoto, Yoshiaki

    1987-06-01

    The performance of a transportable methane stabilized He-Ne laser system, developed for a wavelength-optical frequency standard according to the 1983 Comite Consultatif pour la Definition du Metier, is discussed. An offset-locked laser system using a phase comparison technique is described which is used to evaluate the stabilized laser system. A frequency stability of 2.5 x 10 to the -12th tau exp -1/2, and a resettability of 1 x 10 to the -11th, are estimated for the stabilized laser system.

  6. The relational database system of KM3NeT

    Science.gov (United States)

    Albert, Arnauld; Bozza, Cristiano

    2016-04-01

    The KM3NeT Collaboration is building a new generation of neutrino telescopes in the Mediterranean Sea. For these telescopes, a relational database is designed and implemented for several purposes, such as the centralised management of accounts, the storage of all documentation about components and the status of the detector and information about slow control and calibration data. It also contains information useful during the construction and the data acquisition phases. Highlights in the database schema, storage and management are discussed along with design choices that have impact on performances. In most cases, the database is not accessed directly by applications, but via a custom designed Web application server.

  7. II Lääne-Eesti turismi aastakonverents

    Index Scriptorium Estoniae

    2014-01-01

    MTÜ Lääne-Eesti Turismi ja EASi turismiarenduskeskuse poolt 28. novembril Pärnus Strand SPA & Konverentsihotellis korraldatud aastakonverentsist. Ettekannetega esinesid Kaubandustalituse spetsialist Martti Kalvik, EASi turismiarenduskeskuse direktor Tarmo Mutso, riigikogulane Annely Akkermann, Statistikaameti osakonnajuhataja Ene Saareoja, Mainegrupp OÜ müügijuht Tanel Lips, EAS turismiarenduskeskuse Vene sihtturu juht Agnia Nast, Reisibüroo Pilgrim direktor Igor Paschuk Peterburist, N&A Communications direktor Andrei Petrov Peterburist, Hotell Jurmala SPA väikeaktsionär Antti Aru, Tervis Spaa Grupp juhatuse liige Jaan Ratnik ning konverentsi modereeris Piret Hallik-Sass

  8. 2136-IJBCS-Article-Cesar Bassène

    African Journals Online (AJOL)

    hp

    surface de relevé avant de délimiter l'aire du relevé qui est supérieure ou égale à l'aire minimale. Identification et nomenclature des espèces. Les identifications des espèces ont été ...... des espèces de milieu humide, car les champs sont parfois inondés en plaine saison des pluies tandis que les zones de pâturages ne le.

  9. European Energy Law Seminar 2005. Report of NeVER

    International Nuclear Information System (INIS)

    Oosterom, A.R.; Boumans, L.

    2005-01-01

    An overview is given of the lectures and presentations at the title seminar, which was held in Noordwijk aan Zee, Netherlands, 30-31 May 2005. The seminar was organized by the Dutch Association for Energy Law (NeVER), the Scandinavian Institute for Maritime Law of the University of Oslo, and the Groningen University. The subjects presented concerned recent developments with regard to the internal (European) energy market, LNG, developments in the North Sea area, supply security and quality in a competitive market, reorganization of the European market for natural gas in the light of the liberalization process and privatization of the energy sector [nl

  10. He-, Ne-, and Ar-phosgene intermolecular potential energy surfaces

    DEFF Research Database (Denmark)

    Munteanu, Cristian R.; Henriksen, Christian; Felker, Peter M.

    2013-01-01

    Using the CCSD(T) model, we evaluated the intermolecular potential energy surfaces of the He-, Ne-, and Ar-phosgene complexes. We considered a representative number of intermolecular geometries for which we calculated the corresponding interaction energies with the augmented (He complex) and double......-phosgene surfaces were found to have absolute minima of -72.1, -140.4, and -326.6 cm -1 at distances between the rare-gas atom and the phosgene center of mass of 3.184, 3.254, and 3.516 Å, respectively. The potentials were further used in the evaluation of rovibrational states and the rotational constants...

  11. Retrouvailles avec Hélène Lenoir

    Directory of Open Access Journals (Sweden)

    Lucía Montaner Sánchez

    2012-01-01

    Full Text Available The goal of this interview is to establish a first approach to the work of Hélène Lenoir, a contemporary and relatively unknown to the public at large author. We will explain her first steps as a writer, go through her work amongst the French literary scenario and provide an overview of the more foremost influences received from other authors. The main topical and stylistic features of her literary work are equally reflected, putting special stress on the problematics of tortuous relationships as the orga¬nizing thread of her narrative work.

  12. PLESNA VZGOJA IN RAZLIČNE GLASBENE ZVRSTI

    OpenAIRE

    Trstenjak, Simona

    2016-01-01

    Namen diplomskega dela Plesna vzgoja in različne glasbene zvrsti je s pomočjo teoretičnih izhodišč, ob opazovanju plesnega izražanja otrok, ugotoviti vpliv posamezne glasbene zvrsti na otroka pri plesni vzgoji. Diplomsko delo je sestavljeno iz dveh delov, teoretičnega in empiričnega. V teoretičnem delu smo predstavili teorijo plesne vzgoje v predšolskem obdobju, kjer smo podrobneje opisali sam pomen in vpliv plesne vzgoje na predšolskega otroka, vsebine, cilje in načela ter metode plesne vzgo...

  13. Mass distribution in 20Ne+232Th reaction

    International Nuclear Information System (INIS)

    Sodaye, Suparna; Tripathi, R.; Sudarshan, K.

    2011-01-01

    Mass distribution was measured in 20 Ne+ 232 Th reaction at E lab =145 MeV using recoil catcher technique followed by off-line gamma-ray spectrometry. Significant contribution from transfer fission was observed in the yield of comparatively neutron rich fission products. The variance of mass distribution for complete fusion fission, obtained by excluding neutron rich fission products, was observed to be consistent with the values reported in literature for similar reaction systems which showed a deviation from the systematics obtained using random neck rupture and liquid drop model. (author)

  14. Coulomb breakup of 31Ne using finite range DWBA

    International Nuclear Information System (INIS)

    Shubhchintak; Chatterjee, R.

    2013-01-01

    Coulomb breakup of nuclei away from the valley of stability have been one of the most successful probes to unravel their structure. However, it is only recently that one is venturing into medium mass nuclei like 23 O and 31 Ne. This is a very new and exciting development which has expanded the field of light exotic nuclei to the deformed medium mass region. In this contribution, an extension of the previously proposed theory of Coulomb breakup within the post-form finite range distorted wave Born approximation to include deformation of the projectile is reported

  15. Retention and regeneration of native NAD(H) in noncharged ultrafiltration membrane reactors: application to L-lactate and gluconate production.

    Science.gov (United States)

    Obón, J M; Manjón, A; Iborra, J L

    1998-03-05

    NAD(H) was retained in a noncharged ultrafiltration membrane reactor for the simultaneous and continuous production of L-lactate and gluconate with coenzyme regeneration. Polyethyleneimine (PEI), a 50-kDa cationic polymer, achieved coenzyme retentions above 0.8 for PEI/NAD(H) molar ratios higher than 5. The ionic strength of the inlet medium caused a decrease of NAD(H) retention that can be counterbalanced by an initial addition of 1% bovine serum albumin (BSA). Continuous reactor performance in the presence of PEI and BSA showed that NAD(H), glucose dehydrogenase, and lactate dehydrogenase were retained by 10-kDa ultrafiltration membranes; L-lactate and gluconate were produced at conversions higher than 95%. PEI enhanced the thermal stability of the enzymes used and increased the catalytic efficiency of glucose dehydrogenase, while no effect was found on the kinetic parameters of lactate dehydrogenase. A model that implements the kinetic equations of the two enzymes describes the reactor behavior satisfactorily. In brief, the use of PEI to retain NAD(H) is a new interesting approach to be widely applied in continuous synthesis with the large number of known dehydrogenases. Copyright 1998 John Wiley & Sons, Inc.

  16. [Redox-state of NAD pairs and activity of lactatedehydrogenase and NADase in guinea pig tissues at different stages of development of experimental allergic encephalomyelitis].

    Science.gov (United States)

    Pasichna, E P; Morozova, R P; Donchenko, H V; Chekhivs'ka, L I

    2004-01-01

    Changes of NAD content, redox-state, enzyme activity in the brain and liver tissues of Guinea pigs at different stages of development of experimental allergic encephalomyelitis (EAE) were investigated using the model of multiple sclerosis. It was shown, that the most legible changes of the investigated parameters occur on the preclinical stage and the stage of initial neurological symptoms. The increase of the brain NAD level and reduction properties of NAD+/NADH pairs reduction properties against a background of inhibition of lactatedehydrogenase activity was observed in the early terms of EAE development (7-15 day). The liver lactatedehydrogenase activity is increased at an initial stage. NAD-ase activity is increased in the medium term (18-21 day) that correlates with changes of this enzyme activity in the blood serum. In the term of strongly expressed neurological signs (26-33 day) the sharp drop of NAD content in the brain and liver is observed. The role of the obtained results at different stages of EAE development is discussed.

  17. Half-sandwich rhodium(III) transfer hydrogenation catalysts: Reduction of NAD(+) and pyruvate, and antiproliferative activity.

    Science.gov (United States)

    Soldevila-Barreda, Joan J; Habtemariam, Abraha; Romero-Canelón, Isolda; Sadler, Peter J

    2015-12-01

    Organometallic complexes have the potential to behave as catalytic drugs. We investigate here Rh(III) complexes of general formula [(Cp(x))Rh(N,N')(Cl)], where N,N' is ethylenediamine (en), 2,2'-bipyridine (bpy), 1,10-phenanthroline (phen) or N-(2-aminoethyl)-4-(trifluoromethyl)benzenesulfonamide (TfEn), and Cp(x) is pentamethylcyclopentadienyl (Cp*), 1-phenyl-2,3,4,5-tetramethylcyclopentadienyl (Cp(xPh)) or 1-biphenyl-2,3,4,5-tetramethyl cyclopentadienyl (Cp(xPhPh)). These complexes can reduce NAD(+) to NADH using formate as a hydride source under biologically-relevant conditions. The catalytic activity decreased in the order of N,N-chelated ligand bpy > phen > en with Cp* as the η(5)-donor. The en complexes (1-3) became more active with extension to the Cp(X) ring, whereas the activity of the phen (7-9) and bpy (4-6) compounds decreased. [Cp*Rh(bpy)Cl](+) (4) showed the highest catalytic activity, with a TOF of 37.4±2h(-1). Fast hydrolysis of the chlorido complexes 1-10 was observed by (1)H NMR (hydrogenation reactions was highly dependent on the nature of the chelating ligand and the Cp(x) ring. Competition reactions between NAD(+) and pyruvate for reduction by formate catalysed by 4 showed a preference for reduction of NAD(+). The antiproliferative activity of complex 3 towards A2780 human ovarian cancer cells increased by up to 50% when administered in combination with non-toxic doses of formate, suggesting that transfer hydrogenation can induce reductive stress in cancer cells. Copyright © 2015 The Authors. Published by Elsevier Inc. All rights reserved.

  18. Role of xanthine oxidoreductase and NAD(P)H oxidase in endothelial superoxide production in response to oscillatory shear stress

    Science.gov (United States)

    McNally, J. Scott; Davis, Michael E.; Giddens, Don P.; Saha, Aniket; Hwang, Jinah; Dikalov, Sergey; Jo, Hanjoong; Harrison, David G.

    2003-01-01

    Oscillatory shear stress occurs at sites of the circulation that are vulnerable to atherosclerosis. Because oxidative stress contributes to atherosclerosis, we sought to determine whether oscillatory shear stress increases endothelial production of reactive oxygen species and to define the enzymes responsible for this phenomenon. Bovine aortic endothelial cells were exposed to static, laminar (15 dyn/cm2), and oscillatory shear stress (+/-15 dyn/cm2). Oscillatory shear increased superoxide (O2.-) production by more than threefold over static and laminar conditions as detected using electron spin resonance (ESR). This increase in O2*- was inhibited by oxypurinol and culture of endothelial cells with tungsten but not by inhibitors of other enzymatic sources. Oxypurinol also prevented H2O2 production in response to oscillatory shear stress as measured by dichlorofluorescin diacetate and Amplex Red fluorescence. Xanthine-dependent O2*- production was increased in homogenates of endothelial cells exposed to oscillatory shear stress. This was associated with decreased xanthine dehydrogenase (XDH) protein levels and enzymatic activity resulting in an elevated ratio of xanthine oxidase (XO) to XDH. We also studied endothelial cells lacking the p47phox subunit of the NAD(P)H oxidase. These cells exhibited dramatically depressed O2*- production and had minimal XO protein and activity. Transfection of these cells with p47phox restored XO protein levels. Finally, in bovine aortic endothelial cells, prolonged inhibition of the NAD(P)H oxidase with apocynin decreased XO protein levels and prevented endothelial cell stimulation of O2*- production in response to oscillatory shear stress. These data suggest that the NAD(P)H oxidase maintains endothelial cell XO levels and that XO is responsible for increased reactive oxygen species production in response to oscillatory shear stress.

  19. The 20Ne(d,p)21Ne Transfer Reaction in Relation to the s-Process Abundances

    Energy Technology Data Exchange (ETDEWEB)

    Nsangu, C. T. [University of York, Heslington, YO10 5DD, United Kingdom; Laird, A. M. [University of York, Heslington, YO10 5DD, United Kingdom; Parikh, A. [Univ. Politecnica de Catalana, Spain & Inst. d' Estudis Espacials de Catalunya, Spain; Adsley, P. [University of York, Heslington, YO10 5DD, United Kingdom; Birch, M. D. [McMaster University, Hamilton, Ontario, Canada; Chen, A. A, [McMaster University, Hamilton, Ontario, Canada; Faestermann, T. [Maier-Leibnitz Lab. der Münchner Univ., Garching, Germany; Fox, S. P. [University of York, Heslington, YO10 5DD, United Kingdom; Fulton, B. R. [University of York, Heslington, YO10 5DD, United Kingdom; Hertenberger, R. [Maier-Leibnitz Lab. der Münchner Univ., Garching, Germany; Irvine, D. [McMaster University, Hamilton, Ontario, Canada; Kay, B. P. [University of York, Heslington, YO10 5DD, United Kingdom; Longland, R. [Universitat Politecnica de Catalana, Barcelona, Spain; Manwell, S. [McMaster University, Hamilton, Ontario, Canada; Murphy, A. St. J. [University of Edinburgh, UK; Schmitt, Kyle [ORNL; Séréville, N. de [Institut de Physique Nucléaire d' Orsay & Université Paris Sud 11, Orsay,France; Tomlinson, J. R. [University of York, Heslington, YO10 5DD, United Kingdom; Wirth, H.-F. [Maier-Leibnitz Lab. der Münchner Univ., Garching, Germany

    2016-01-01

    A study of the Ne-20(d,p)Ne-21 transfer reaction was performed using the Quadrupole Dipole Dipole Dipole (Q3D) magnetic spectrograph in Garching, Germany. The experiment probed excitation energies in Ne-21 ranging from 6.9 MeV to 8.5 MeV. The aim was to investigate the spectroscopic information of Ne-21 within the Gamow window of core helium burning in massive stars. Further information in this region will help reduce the uncertainties on the extrapolation down to Gamow window cross sections of the O-17(alpha,gamma)Ne-21 reaction. In low metallicity stars, this reaction has a direct impact on s-process abundances by determining the fate of O-16 as either a neutron poison or a neutron absorber. The experiment used a 22-MeV deuteron beam, with intensities varying from 0.5-1 mu A, and an implanted target of Ne-20 of 7 mu g/cm(2) in 40 mu g/cm(2) carbon foils. Sixteen Ne-21 peaks have been identified in the E-x = 6.9-8.5 MeV range, of which only thirteen peaks correspond to known states. Only the previously-known E-x = 7.960 MeV state was observed within the Gamow window.

  20. Absorbed dose to water comparison between NE 2561 and NE 2671 chambers using IAEA, HPA and NACP protocols for gamma ray beam

    International Nuclear Information System (INIS)

    Mohd Taufik Dolah; Noriah Mod Ali; Taiman Kadni

    2002-01-01

    The aim of this study to evaluate the performance of NE 2571 chamber in comparison with NE 2561 chamber in determination of the absorbed dose to water in gamma ray beam. In this study NE 2561 is taking as a reference standard chamber while NE 2571 as a working standard. Irradiation of chamber (alternately) was performed at a reference depth, 5 cm, inside the IAEA water phantom. Both chambers were exposed to 13 difference exposures of gamma rays. The values of absorbed dose to water were then determined using IAEA, HPA and NACP protocols. Deviations of absorbed dose determined by NE 2561 and NE 2571 were calculated for each protocol. result obtained in terms of [protocol, μ (mean deviation) ± σ s e (standard error)] were (IAEA, 1.12 ± 0.04], [HPA, 0.09 ± 0.04], and [NCP, 0.09 ± 0.04]. It can be concluded that NE 2571 shown acceptable performance as it is within acceptable limit ± 3%. (Author)

  1. Lääne-Virumaa ettevõtete TOP aastal 2004

    Index Scriptorium Estoniae

    2005-01-01

    Tabelid: Lääne-Virumaa ettevõtete TOP 50; Käibe TOP 35; Kasumi TOP 35; Lääne-Virumaa ettevõtete üld- ja finantsandmed; Käibe kasvu TOP 20; Kasumi kasvu TOP 20; Rentaabluse TOP 20; Omakapitali tootlikkuse TOP 20. Vt. samas: Viktor Sepp, Merike Lees. Lääne-Virumaal üllatavad uued tegijad

  2. Le facteur temps ne sonne jamais deux fois

    CERN Document Server

    Klein, Etienne

    2009-01-01

    Chose déroutante, décidément, que le temps. Nous en parlons comme d'une notion familière, évidente, voire domestique, "gérable". Nous parlons même d'un "temps réel" pour évoquer l'instantanéité, c'est-à-dire le temps sur lequel nous n'avons aucune prise. Les physiciens, eux, l'ont couplé à l'espace, en ont fait une variable mathématique, abstraite, qu'ils intègrent dans des théories audacieuses, spectaculaires, si complexes qu'elles sont difficiles à traduire en langage courant. Certains disent même avoir identifié le moteur du temps. Quant aux philosophes, ils ne cessent depuis plus de deux millénaires de soumettre le temps au questionnement : est-il une sorte d'entité primitive, originaire, qui ne dériverait que d'elle-même? Ou procéderait-il au contraire d'une ou plusieurs autres entités, plus fondamentales: la relation de cause à effet, par exemple? Le temps s'écoule-t-il de lui-même ou a-t-il besoin des événements qui s'y déroulent pour passer? S'apparente-t-il au devenir,...

  3. Radiometric ages of the Akashima Formation, Oga Peninsula, NE Japan

    International Nuclear Information System (INIS)

    Kano, Kazuhiko; Ishizuka, Osamu; Tani, Kenichiro; Iwano, Hideki; Danhara, Tohru; Ohguchi, Takeshi; Dunkley, Daniel J.

    2012-01-01

    U-Pb and fission-track dating of zircons and Ar-Ar dating of plagioclase were conducted to estimate the eruption age of the welded dacite lapilli-tuff of the Akashima Formation, Oga Peninsula, NE Japan, yielding ages of 72 Ma, 65-63 Ma and 34 Ma, respectively. The zircon grains are mainly euhedral with zonal structures concordant with their external form, and are set in a welded matrix and in dense lenses of pumice; thus, they are likely to be juvenile. The Standard deviation of U-Pb ages of different zircon grains is small (1-2 Ma) and their age likely represents the eruption age of the magma. Fission-track ages are significantly younger than U-Pb ages, and are thought to represent thermally rejuvenation as suggested by shortening of fission tracks. The Ar-Ar age of the plagioclase remains fairly stable at almost all stages of heating; the plateau age, however, falls within the time range of the Late Eocene Monzen Formation and could represent almost complete albitization ages by associated volcanic activity. The U-Pb ages provide reliable evidence for presence of late Late Cretaceous acidic volcanism in NE Japan. (author)

  4. Biphasic kinetic behavior of E. coli WrbA, an FMN-dependent NAD(PH:quinone oxidoreductase.

    Directory of Open Access Journals (Sweden)

    Iryna Kishko

    Full Text Available The E. coli protein WrbA is an FMN-dependent NAD(PH:quinone oxidoreductase that has been implicated in oxidative defense. Three subunits of the tetrameric enzyme contribute to each of four identical, cavernous active sites that appear to accommodate NAD(PH or various quinones, but not simultaneously, suggesting an obligate tetramer with a ping-pong mechanism in which NAD departs before oxidized quinone binds. The present work was undertaken to evaluate these suggestions and to characterize the kinetic behavior of WrbA. Steady-state kinetics results reveal that WrbA conforms to a ping-pong mechanism with respect to the constancy of the apparent Vmax to Km ratio with substrate concentration. However, the competitive/non-competitive patterns of product inhibition, though consistent with the general class of bi-substrate reactions, do not exclude a minor contribution from additional forms of the enzyme. NMR results support the presence of additional enzyme forms. Docking and energy calculations find that electron-transfer-competent binding sites for NADH and benzoquinone present severe steric overlap, consistent with the ping-pong mechanism. Unexpectedly, plots of initial velocity as a function of either NADH or benzoquinone concentration present one or two Michaelis-Menten phases depending on the temperature at which the enzyme is held prior to assay. The effect of temperature is reversible, suggesting an intramolecular conformational process. WrbA shares these and other details of its kinetic behavior with mammalian DT-diaphorase, an FAD-dependent NAD(PH:quinone oxidoreductase. An extensive literature review reveals several other enzymes with two-plateau kinetic plots, but in no case has a molecular explanation been elucidated. Preliminary sedimentation velocity analysis of WrbA indicates a large shift in size of the multimer with temperature, suggesting that subunit assembly coupled to substrate binding may underlie the two-plateau behavior. An

  5. Recombinant NAD-dependent SIR-2 protein of Leishmania donovani: immunobiochemical characterization as a potential vaccine against visceral leishmaniasis.

    Science.gov (United States)

    Baharia, Rajendra K; Tandon, Rati; Sharma, Tanuj; Suthar, Manish K; Das, Sanchita; Siddiqi, Mohammad Imran; Saxena, Jitendra Kumar; Sundar, Shaym; Sunder, Shyam; Dube, Anuradha

    2015-03-01

    The development of a vaccine conferring long-lasting immunity remains a challenge against visceral leishmaniasis (VL). Immunoproteomic characterization of Leishmania donovani proteins led to the identification of a novel protein NAD+-dependent Silent Information regulatory-2 (SIR2 family or sirtuin) protein (LdSir2RP) as one of the potent immunostimulatory proteins. Proteins of the SIR2 family are characterized by a conserved catalytic domain that exerts unique NAD-dependent deacetylase activity. In the present study, an immunobiochemical characterization of LdSir2RP and further evaluation of its immunogenicity and prophylactic potential was done to assess for its possible involvement as a vaccine candidate against leishmaniasis. LdSir2RP was successfully cloned, expressed and purified. The gene was present as a monomeric protein of ~45 kDa and further established by the crosslinking experiment. rLdSir2RP shown cytosolic localization in L. donovani and demonstrating NAD+-dependent deacetylase activity. Bioinformatic analysis also confirmed that LdSir2RP protein has NAD binding domain. The rLdSir2RP was further assessed for its cellular response by lymphoproliferative assay and cytokine ELISA in cured Leishmania patients and hamsters (Mesocricetus auratus) in comparison to soluble Leishmania antigen and it was observed to stimulate the production of IFN-γ, IL-12 and TNF-α significantly but not the IL-4 and IL-10. The naïve hamsters when vaccinated with rLdSir2RP alongwith BCG resisted the L. donovani challenge to the tune of ~75% and generated strong IL-12 and IFN-γ mediated Th1 type immune response thereof. The efficacy was further supported by remarkable increase in IgG2 antibody level which is indicative of Th1 type of protective response. Further, with a possible implication in vaccine design against VL, identification of potential T-cell epitopes of rLdSir2RP was done using computational approach. The immunobiochemical characterization strongly suggest the

  6. The current state of forests in the lower reaches of the Vah section Sala - Nove Mesto nad Vahom

    International Nuclear Information System (INIS)

    Abrahamova, A.

    2009-01-01

    Area of interest lies in the central part of the Danube plain and extends along both sides of the river Vah. Naturally occurring habitats close to small-scale, and are considerable anthropicly contingent either directly (forestry) or indirectly (modified water regime). The contribution gives phyto-sociological characteristics of forests in the lower reaches of the Vah section Sala - Nove Mesto nad Vahom. We drew up the 21 phyto-sociological entries characterizing forest vegetation, which we classified into two associations Salici-Populetum, Fraxino Ulmetum-and community-Crataegus monogyna Populus nigra.

  7. Monte Carlo studies of the KM3NeT physics performance

    Science.gov (United States)

    Shanidze, Rezo; Kuch, Sebastian; Katz, Ulrich

    2009-04-01

    KM3NeT neutrino telescope configurations with different detector components and geometry have been simulated and studied with modified ANTARES software. The physics performance of KM3NeT is characterised by two parameters: neutrino effective area and angular resolution of the reconstructed muons. These two benchmark parameters are determined and compared for the different KM3NeT options simulated. The physics performance of the KM3NeT configuration is evaluated by calculating achievable average upper limits for cosmic neutrino fluxes.

  8. Monte Carlo studies of the KM3NeT physics performance

    Energy Technology Data Exchange (ETDEWEB)

    Shanidze, Rezo [Erlangen Centre for Astroparticle Physics (ECAP), University of Erlangen-Nuernberg, Erwin-Rommel-Street 1, D-91058 Erlangen (Germany)], E-mail: shanidze@physik.uni-erlangen.de; Kuch, Sebastian; Katz, Ulrich [Erlangen Centre for Astroparticle Physics (ECAP), University of Erlangen-Nuernberg, Erwin-Rommel-Street 1, D-91058 Erlangen (Germany)

    2009-04-11

    KM3NeT neutrino telescope configurations with different detector components and geometry have been simulated and studied with modified ANTARES software. The physics performance of KM3NeT is characterised by two parameters: neutrino effective area and angular resolution of the reconstructed muons. These two benchmark parameters are determined and compared for the different KM3NeT options simulated. The physics performance of the KM3NeT configuration is evaluated by calculating achievable average upper limits for cosmic neutrino fluxes.

  9. Room temperature wafer direct bonding of smooth Si surfaces recovered by Ne beam surface treatments

    Science.gov (United States)

    Kurashima, Yuichi; Maeda, Atsuhiko; Takagi, Hideki

    2013-06-01

    We examined the applicability of a Ne fast atom beam (FAB) to surface activated bonding of Si wafers at room temperature. With etching depth more than 1.5 nm, the bonding strength comparable to Si bulk strength was attained. Moreover, we found the improvement of the bonding strength by surface smoothing effect of the Ne FAB. Silicon surface roughness decreased from 0.40 to 0.17 nm rms by applying a Ne FAB of 30 nm etching depth. The bonding strength between surfaces recovered by Ne FAB surface smoothing was largely improved and finally became equivalent to Si bulk strength.

  10. UV luminescence of NeD in solid neon-deuterium mixtures

    DEFF Research Database (Denmark)

    Stenum, B.; Schou, Jørgen; Gürtler, P.

    1994-01-01

    Solid samples of neon-deuterium mixtures were irradiated by keV electrons, and the luminescence was measured between 100 and 300 nm. For concentrations between 0.1% D-2 in Ne and 1% Ne in D-2 an intense emission band was observed. The maximum intensity was observed for 10% D-2 in Ne. Comparisons...... with results from gas phase measurements indicate that the dominant component of the band originates from a bound-free transition from the A(2) Sigma(+) state of NeD to the repulsive ground state....

  11. Naine kohtub Hundiga - püüne peal / Kaja Kann, Bush Hartshorn

    Index Scriptorium Estoniae

    Kann, Kaja, 1973-

    2006-01-01

    Kanuti Gildi saalis etendub 26.-28. veebr. Kaja Kanni ja Bush Hartshorni lavaduett "Naine ja Hunt". Autorid räägivad, kuidas nad käisid Pärnumaa laantes oma lavaloole õiget tunnetust otsimas. Lisatud andmed Bush Hartshorni kohta

  12. Identification of NAD(PH quinone oxidoreductase activity in azoreductases from P. aeruginosa: azoreductases and NAD(PH quinone oxidoreductases belong to the same FMN-dependent superfamily of enzymes.

    Directory of Open Access Journals (Sweden)

    Ali Ryan

    Full Text Available Water soluble quinones are a group of cytotoxic anti-bacterial compounds that are secreted by many species of plants, invertebrates, fungi and bacteria. Studies in a number of species have shown the importance of quinones in response to pathogenic bacteria of the genus Pseudomonas. Two electron reduction is an important mechanism of quinone detoxification as it generates the less toxic quinol. In most organisms this reaction is carried out by a group of flavoenzymes known as NAD(PH quinone oxidoreductases. Azoreductases have previously been separate from this group, however using azoreductases from Pseudomonas aeruginosa we show that they can rapidly reduce quinones. Azoreductases from the same organism are also shown to have distinct substrate specificity profiles allowing them to reduce a wide range of quinones. The azoreductase family is also shown to be more extensive than originally thought, due to the large sequence divergence amongst its members. As both NAD(PH quinone oxidoreductases and azoreductases have related reaction mechanisms it is proposed that they form an enzyme superfamily. The ubiquitous and diverse nature of azoreductases alongside their broad substrate specificity, indicates they play a wide role in cellular survival under adverse conditions.

  13. Small kernel 1 encodes a pentatricopeptide repeat protein required for mitochondrial nad7 transcript editing and seed development in maize (Zea mays) and rice (Oryza sativa).

    Science.gov (United States)

    Li, Xiao-Jie; Zhang, Ya-Feng; Hou, Mingming; Sun, Feng; Shen, Yun; Xiu, Zhi-Hui; Wang, Xiaomin; Chen, Zong-Liang; Sun, Samuel S M; Small, Ian; Tan, Bao-Cai

    2014-09-01

    RNA editing modifies cytidines (C) to uridines (U) at specific sites in the transcripts of mitochondria and plastids, altering the amino acid specified by the DNA sequence. Here we report the identification of a critical editing factor of mitochondrial nad7 transcript via molecular characterization of a small kernel 1 (smk1) mutant in Zea mays (maize). Mutations in Smk1 arrest both the embryo and endosperm development. Cloning of Smk1 indicates that it encodes an E-subclass pentatricopeptide repeat (PPR) protein that is targeted to mitochondria. Loss of SMK1 function abolishes the C → U editing at the nad7-836 site, leading to the retention of a proline codon that is edited to encode leucine in the wild type. The smk1 mutant showed dramatically reduced complex-I assembly and NADH dehydrogenase activity, and abnormal biogenesis of the mitochondria. Analysis of the ortholog in Oryza sativa (rice) reveals that rice SMK1 has a conserved function in C → U editing of the mitochondrial nad7-836 site. T-DNA knock-out mutants showed abnormal embryo and endosperm development, resulting in embryo or seedling lethality. The leucine at NAD7-279 is highly conserved from bacteria to flowering plants, and analysis of genome sequences from many plants revealed a molecular coevolution between the requirement for C → U editing at this site and the existence of an SMK1 homolog. These results demonstrate that Smk1 encodes a PPR-E protein that is required for nad7-836 editing, and this editing is critical to NAD7 function in complex-I assembly in mitochondria, and hence to embryo and endosperm development in maize and rice. © 2014 The Authors The Plant Journal © 2014 John Wiley & Sons Ltd.

  14. Relative importance of redox buffers GSH and NAD(P)H in age-related neurodegeneration and Alzheimer disease-like mouse neurons.

    Science.gov (United States)

    Ghosh, Debolina; Levault, Kelsey R; Brewer, Gregory J

    2014-08-01

    Aging, a major risk factor in Alzheimer's disease (AD), is associated with an oxidative redox shift, decreased redox buffer protection, and increased free radical reactive oxygen species (ROS) generation, probably linked to mitochondrial dysfunction. While NADH is the ultimate electron donor for many redox reactions, including oxidative phosphorylation, glutathione (GSH) is the major ROS detoxifying redox buffer in the cell. Here, we explored the relative importance of NADH and GSH to neurodegeneration in aging and AD neurons from nontransgenic and 3xTg-AD mice by inhibiting their synthesis to determine whether NADH can compensate for the GSH loss to maintain redox balance. Neurons stressed by either depleting NAD(P)H or GSH indicated that NADH redox control is upstream of GSH levels. Further, although depletion of NAD(P)H or GSH correlated linearly with neuron death, compared with GSH depletion, higher neurodegeneration was observed when NAD(P)H was extrapolated to zero, especially in old age, and in the 3xTg-AD neurons. We also observed an age-dependent loss of gene expression of key redox-dependent biosynthetic enzymes, NAMPT (nicotinamide phosphoribosyltransferase), and NNT (nicotinamide nucleotide transhydrogenase). Moreover, age-related correlations between brain NNT or NAMPT gene expression and NADPH levels suggest that these genes contribute to the age-related declines in NAD(P)H. Our data indicate that in aging and more so in AD-like neurons, NAD(P)H redox control is upstream of GSH and an oxidative redox shift that promotes neurodegeneration. Thus, NAD(P)H generation may be a more efficacious therapeutic target upstream of GSH and ROS. © 2014 The Authors. Aging Cell published by the Anatomical Society and John Wiley & Sons Ltd.

  15. Miocene uplift of the NE Greenland margin linked to plate tectonics: Seismic evidence from the Greenland Fracture Zone, NE Atlantic

    DEFF Research Database (Denmark)

    Døssing Andreasen, Arne; Japsen, Peter; Watts, Anthony B.

    2016-01-01

    Tectonic models predict that, following breakup, rift margins undergo only decaying thermal subsidence during their post-rift evolution. However, post-breakup stratigraphy beneath the NE Atlantic shelves shows evidence of regional-scale unconformities, commonly cited as outer margin responses to ...... by plate tectonic forces, induced perhaps by a change in the Iceland plume (a hot pulse) and/or by changes in intra-plate stresses related to global tectonics.......Tectonic models predict that, following breakup, rift margins undergo only decaying thermal subsidence during their post-rift evolution. However, post-breakup stratigraphy beneath the NE Atlantic shelves shows evidence of regional-scale unconformities, commonly cited as outer margin responses...... backstripping. We explain the thermo-mechanical coupling and the deposition of contourites by the formation of a continuous plate boundary along the Mohns and Knipovich ridges, leading to an accelerated widening of the Fram Strait. We demonstrate that the IMU event is linked to onset of uplift and massive shelf...

  16. Tryptophan phosphorescence spectroscopy reveals that a domain in the NAD(H)-binding component (dI) of transhydrogenase from Rhodospirillum rubrum has an extremely rigid and conformationally homogeneous protein core

    NARCIS (Netherlands)

    Broos, J; Gabellieri, E; van Boxel, GI; Jackson, JB; Strambini, GB; Strambini, Giovanni B.

    2003-01-01

    The characteristics of tryptophan phosphorescence from the NAD(H)-binding component (dI) component of Rhodospirillum rubrum transhydrogenase are described. This enzyme couples hydride transfer between NAD( H) and NADP( H) to proton translocation across a membrane and is only active as a dimer.

  17. An improved glycerol biosensor with an Au-FeS-NAD-glycerol-dehydrogenase anode.

    Science.gov (United States)

    Mahadevan, Aishwarya; Fernando, Sandun

    2017-06-15

    An improved glycerol biosensor was developed via direct attachment of NAD + -glycerol dehydrogenase coenzyme-apoenzyme complex onto supporting gold electrodes, using novel inorganic iron (II) sulfide (FeS)-based single molecular wires. Sensing performance factors, i.e., sensitivity, a detection limit and response time of the FeS and conventional pyrroloquinoline quinone (PQQ)-based biosensor were evaluated by dynamic constant potential amperometry at 1.3V under non-buffered conditions. For glycerol concentrations ranging from 1 to 25mM, a 77% increase in sensitivity and a 53% decrease in detection limit were observed for the FeS-based biosensor when compared to the conventional PQQ-based counterpart. The electrochemical behavior of the FeS-based glycerol biosensor was analyzed at different concentrations of glycerol, accompanied by an investigation into the effects of applied potential and scan rate on the current response. Effects of enzyme stimulants ((NH 4 ) 2 SO 4 and MnCl 2 ·4H 2 O) concentrations and buffers/pH (potassium phosphate buffer pH 6-8, Tris buffer pH 8-10) on the current responses generated by the FeS-based glycerol biosensor were also studied. The optimal detection conditions were 0.03M (NH 4 ) 2 SO 4 and 0.3µm MnCl 2 ·4H 2 O in non-buffered aqueous electrolyte under stirring whereas under non-stirring, Tris buffer at pH 10 with 0.03M (NH 4 ) 2 SO 4 and 30µm MnCl 2 ·4H 2 O were found to be optimal detection conditions. Interference by glucose, fructose, ethanol, and acetic acid in glycerol detection was studied. The observations indicated a promising enhancement in glycerol detection using the novel FeS-based glycerol sensing electrode compared to the conventional PQQ-based one. These findings support the premise that FeS-based bioanodes are capable of biosensing glycerol successfully and may be applicable for other enzymatic biosensors. Copyright © 2016 Elsevier B.V. All rights reserved.

  18. NAD(PH:quinone oxidoreductase 1 inducer activity of some novel anilinoquinazoline derivatives

    Directory of Open Access Journals (Sweden)

    Ghorab MM

    2016-08-01

    Full Text Available Mostafa M Ghorab,1,2 Mansour S Alsaid,1 Maureen Higgins,3 Albena T Dinkova-Kostova,3–5 Abdelaaty A Shahat,1,6 Nehal H Elghazawy,7 Reem K Arafa7,8 1Department of Pharmacognosy, College of Pharmacy, King Saud University, Riyadh, Kingdom of Saudi Arabia; 2Department of Drug Radiation Research, National Center for Radiation Research & Technology, Atomic Energy Authority, Cairo, Egypt; 3Jacqui Wood Cancer Centre, Division of Cancer Research, Medical Research Institute, University of Dundee, Dundee, UK; 4Department of Medicine, 5Department of Pharmacology and Molecular Sciences, Johns Hopkins University School of Medicine, Baltimore, MD, USA; 6Phytochemistry Department, National Research Center, Dokki, Giza, 7Zewail City of Science and Technology, 8Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Cairo University, Cairo, Egypt Abstract: The Kelch-like ECH-associated protein 1 (Keap1/nuclear factor erythroid 2-related factor 2 (Nrf2/antioxidant response elements pathway enables cells to survive oxidative stress conditions through regulating the expression of cytoprotective enzymes such as NAD(PH:quinone oxidoreductase 1 (NQO1. This work presents the design and synthesis of novel anilinoquinazoline derivatives (2–16a and evaluation of their NQO1 inducer activity in murine cells. Molecular docking of the new compounds was performed to assess their ability to inhibit Keap1–Nrf2 protein–protein interaction through occupying the Keap1–Nrf2-binding domain, which leads to Nrf2 accumulation and enhanced gene expression of NQO1. Docking results showed that all compounds can potentially interact with Keap1; however, 1,5-dimethyl-2-phenyl-4-(2-phenylquinazolin-4-ylamino-1,2-dihydropyrazol-3-one (9, the most potent inducer, showed the largest number of interactions with key amino acids in the binding pocket (Arg483, Tyr525, and Phe478 compared to the native ligand or any other compound in this series. Keywords: Kelch domain, molecular

  19. A substrate in pieces: allosteric activation of glycerol 3-phosphate dehydrogenase (NAD+) by phosphite dianion.

    Science.gov (United States)

    Tsang, Wing-Yin; Amyes, Tina L; Richard, John P

    2008-04-22

    The ratio of the second-order rate constants for reduction of dihydroxyacetone phosphate (DHAP) and of the neutral truncated substrate glycolaldehyde (GLY) by glycerol 3-phosphate dehydrogenase (NAD (+), GPDH) saturated with NADH is (1.0 x 10 (6) M (-1) s (-1))/(8.7 x 10 (-3) M (-1) s (-1)) = 1.1 x 10 (8), which was used to calculate an intrinsic phosphate binding energy of at least 11.0 kcal/mol. Phosphite dianion binds very weakly to GPDH ( K d > 0.1 M), but the bound dianion strongly activates GLY toward enzyme-catalyzed reduction by NADH. Thus, the large intrinsic phosphite binding energy is expressed only at the transition state for the GPDH-catalyzed reaction. The ratio of rate constants for the phosphite-activated and the unactivated GPDH-catalyzed reduction of GLY by NADH is (4300 M (-2) s (-1))/(8.7 x 10 (-3) M (-1) s (-1)) = 5 x 10 (5) M (-1), which was used to calculate an intrinsic phosphite binding energy of -7.7 kcal/mol for the association of phosphite dianion with the transition state complex for the GPDH-catalyzed reduction of GLY. Phosphite dianion has now been shown to activate bound substrates for enzyme-catalyzed proton transfer, decarboxylation, hydride transfer, and phosphoryl transfer reactions. Structural data provide strong evidence that enzymic activation by the binding of phosphite dianion occurs at a modular active site featuring (1) a binding pocket complementary to the reactive substrate fragment which contains all the active site residues needed to catalyze the reaction of the substrate piece or of the whole substrate and (2) a phosphate/phosphite dianion binding pocket that is completed by the movement of flexible protein loop(s) to surround the nonreacting oxydianion. We propose that loop motion and associated protein conformational changes that accompany the binding of phosphite dianion and/or phosphodianion substrates lead to encapsulation of the substrate and/or its pieces in the protein interior, and to placement of the active

  20. Expression, purification, crystallization and preliminary X-ray analysis of an NAD-dependent glyceraldehyde-3-phosphate dehydrogenase from Helicobacter pylori

    International Nuclear Information System (INIS)

    Elliott, Paul R.; Mohammad, Shabaz; Melrose, Helen J.; Moody, Peter C. E.

    2008-01-01

    Glyceraldehyde-3-phosphate dehydrogenase B from H. pylori has been cloned, expressed, purified and crystallized in the presence of NAD. Crystals of GAPDHB diffracted to 2.8 Å resolution and belonged to space group P6 5 22, with unit-cell parameters a = b = 166.1, c = 253.1 Å. Helicobacter pylori is a dangerous human pathogen that resides in the upper gastrointestinal tract. Little is known about its metabolism and with the onset of antibiotic resistance new treatments are required. In this study, the expression, purification, crystallization and preliminary X-ray diffraction of an NAD-dependent glyceraldehyde-3-phosphate dehydrogenase from H. pylori are reported

  1. Perturbations of NAD+ salvage systems impact mitochondrial function and energy homeostasis in mouse myoblasts and intact skeletal muscle

    DEFF Research Database (Denmark)

    Andersen, Marianne Agerholm; Dall, Morten; Jensen, Benjamin Anderschou Holbech

    2018-01-01

    Nampt KD) C2C12 cells using a shRNA lentiviral approach. Moreover, we applied gene electrotransfer to express cre recombinase in tibialis anterior muscle of floxed Nampt mice. In shNampt KD C2C12 myoblasts, Nampt and NAD+ levels were reduced by 70% and 50%, respectively, and maximal respiratory capacity......Nampt KD cells, respectively. Expression of cre recombinase in muscle of floxed Nampt mice reduced NAMPT and NAD+ levels by 38% and 43%, respectively. Glucose uptake increased by 40% and mitochondrial complex IV respiration was compromised by 20%. HIF1α-regulated genes and histone H3 lysine 9 (H3K9...

  2. Udržitelný rozvoj města, v aplikaci na Nové Město nad Metují

    OpenAIRE

    Maur, Vilém

    2014-01-01

    The work deals with the sustainable development of the city, in the Nove Mesto nad Metují. The first part theoretical, is engaged in the development of the concept of sustainable development and presents the research methodology of sustainability of the city. In the practical part, these methods are applied to the sustainable development of the Nove Město nad Metují. Sustainable urban development is studied from four aspects, land use, environmental, economic and social. The aim of the work i...

  3. Efficient Sensor Integration on Platforms (NeXOS)

    Science.gov (United States)

    Memè, S.; Delory, E.; Del Rio, J.; Jirka, S.; Toma, D. M.; Martinez, E.; Frommhold, L.; Barrera, C.; Pearlman, J.

    2016-12-01

    In-situ ocean observing platforms provide power and information transmission capability to sensors. Ocean observing platforms can be mobile, such as ships, autonomous underwater vehicles, drifters and profilers, or fixed, such as buoys, moorings and cabled observatories. The process of integrating sensors on platforms can imply substantial engineering time and resources. Constraints range from stringent mechanical constraints to proprietary communication and control firmware. In NeXOS, the implementation of a PUCK plug and play capability is being done with applications to multiple sensors and platforms. This is complemented with a sensor web enablement that addresses the flow of information from sensor to user. Open standards are being tested in order to assess their costs and benefits in existing and future observing systems. Part of the testing implied open-source coding and hardware prototyping of specific control devices in particular for closed commercial platforms where firmware upgrading is not straightforward or possible without prior agreements or service fees. Some platform manufacturers such as European companies ALSEAMAR[1] and NKE Instruments [2] are currently upgrading their control and communication firmware as part of their activities in NeXOS. The sensor development companies Sensorlab[3] SMID[4] and TRIOS [5]upgraded their firmware with this plug and play functionality. Other industrial players in Europe and the US have been sent NeXOS sensors emulators to test the new protocol on their platforms. We are currently demonstrating that with little effort, it is also possible to have such middleware implemented on very low-cost compact computers such as the open Raspberry Pi[6], and have a full end-to-end interoperable communication path from sensor to user with sensor plug and play capability. The result is an increase in sensor integration cost-efficiency and the demonstration will be used to highlight the benefit to users and ocean observatory

  4. Methanol/Oxygen Enzymatic Biofuel Cell Using Laccase and NAD+-Dependent Dehydrogenase Cascades as Biocatalysts on Carbon Nanodots Electrodes.

    Science.gov (United States)

    Wu, Guozhi; Gao, Yue; Zhao, Dan; Ling, Pinghua; Gao, Feng

    2017-11-22

    The efficient immobilization of enzymes on favorable supporting materials to design enzyme electrodes endowed with specific catalysis performances such as deep oxidation of biofuels, and direct electron transfer (DET)-type bioelectrocatalysis is highly desired for fabricating enzymatic biofuel cells (BFCs). In this study, carbon nanodots (CNDs) have been used as the immobilizing matrixes and electron relays of enzymes to construct (NAD + )-dependent dehydrogenase cascades-based bioanode for the deep oxidation of methanol and DET-type laccase-based biocathode for oxygen reduction to water. At the bioanode, multiplex enzymes including alcohol dehydrogenase, aldehyde dehydrogenase, and formate dehydrogenase are coimmobilized on CNDs electrode which is previously coated with in situ polymerized methylene blue as the electrocatalyst for oxidizing NADH to NAD + . At the biocathode, fungal laccase is directly cast on CNDs and facilitated DET reaction is allowed. As a result, a novel membrane-less methanol/O 2 BFC has been assembled and displays a high open-circuit voltage of 0.71(±0.02) V and a maximum power density of 68.7 (±0.4) μW cm -2 . These investigated features imply that CNDs may act as new conductive architectures to elaborate enzyme electrodes for further bioelectrochemical applications.

  5. Toward the virtual screening of potential drugs in the homology modeled NAD+ dependent DNA ligase from Mycobacterium tuberculosis.

    Science.gov (United States)

    Singh, Vijai; Somvanshi, Pallavi

    2010-02-01

    DNA ligase is an important enzyme and it plays vital role in the replication and repair; also catalyzes nick joining between adjacent bases of DNA. The NAD(+) dependent DNA ligase is selectively present in eubacteria and few viruses; but missing in humans. Homology modeling was used to generate 3-D structure of NAD(+) dependent DNA ligase (LigA) of Mycobacterium tuberculosis using the known template (PDB: 2OWO). Furthermore, the stereochemical quality and torsion angle of 3-D structure was validated. Numerous effective drugs were selected and the active amino acid residue in LigA was targeted and virtual screening through molecular docking was done. In this analysis, four drugs Chloroquine, Hydroxychloroquine, Putrienscine and Adriamycin were found more potent in inhibition of M. tuberculosis through the robust binding affinity between protein-drug interactions in comparison with the other studied drugs. A phylogenetic tree was constructed and it was observed that homology of LigA in M. tuberculosis resembled with other Mycobacterium species. The conserved active amino acids of LigA may be useful to target these drugs. These findings could be used as the starting point of a rational design of novel antibacterial drugs and its analogs.

  6. AKIBAT BENCANA TSUNAMI TERHADAP PELAYANAN KESEHATAN DI PROVINSI NANGGROE ACEH DARUSSALAM (NAD: SUATU KAJIAN CEPAT GUNA MASUKAN BAGI KEBIJAKAN

    Directory of Open Access Journals (Sweden)

    Agus Suwandono

    2012-10-01

    Full Text Available On 26 December 2004, a series of earthquakes followed by tsunami with an epicentrum in Indian Ocean next to the northwest coast of Sumatra Island caused very serious damaged in Nanggroe Aceh Darussalam (NAD and North Sumatra Provinces. Around 100-150 thousand population were predicted to be the victims, either dead, missing or seriously wounded due to this  world biggest disaster in 2004. This disaster also destroyed the local health services and their human resources on health. A team of rapid assessment was peiformed by Ministry of Health (MOH to provide inputs for rehabilitation of the local health services. Observation, interview and document searching were used as the methods of this rapid assessment. Although there were many weaknesses of this rapid assessment, some findings, analysis and recommendations were provided by the team as follows: 1. MOH should take over immediately and responsible to the recovery of the local health services and solved the 10 issues found by the team; 2. The three phases recommended by the team especially for CDC programs should be considered seriously to be carried out after the tsunami; 3. Deeper assessment should be carried after the basic health system in NAD was recovered; and 4. Rebuilding of local HCs and hospitals as well as supplying the human resources on health should be planned and implemented immedietely.

  7. Differential levels of metabolic activity in isolated versus confluent/partially confluent HeLa cells are analyzed by autofluorescent NAD(P)H using multi-photon FLIM microscopy

    Science.gov (United States)

    Chandler, Andrea; Chandler, Aaron; Wallrabe, Horst; Periasamy, Ammasi

    2017-02-01

    NAD(P)H is a known biomarker for cellular metabolism; a higher ratio of enzyme-bound NAD(P)H to free/unbound NAD(P)H indicates an increase in metabolic activity. Free NADH has a shorter fluorescence lifetime (τ1), the bound version (τ2) a longer lifetime. FLIM's unique capability to establish inter alia the relative fractions of τ1 (a1%) and τ2 (a2%) in each pixel, determines the level of metabolic activity. The relative abundances of bound NAD(P)H were analyzed for single cells, confluent and partially confluent cells within 3 Fields-of-View (FoVs). A gradient of increasing a 2% levels of bound NAD(P)H from single, partially confluent to confluent cells was observed.

  8. ‘Transnationalising’ Ne Bis In Idem: How the Rule of Ne Bis In Idem Reveals the Principle of Personal Legal Certainty

    Directory of Open Access Journals (Sweden)

    Juliette Lelieur

    2013-09-01

    Full Text Available Since Article 54 of the Convention implementing the Schengen Agreement gave the rule of ne bis in idem a transnational dimension, talk of the ‘transnational ne bis in idem principle’ has been commonplace. Thus, when looking for general principles of transnational criminal law, scholars refer to the principle of ‘transnational ne bis in idem’. It is doubtful, however, that ne bis in idem qualifies as a principle of law. It should be regarded, rather, as a rule of criminal procedure, traditionally based on the principle of res judicata. Giving the rule of ne bis in idem a transnational dimension therefore requires either transnationalising the principle of res judicata, or giving the rule of ne bis in idem a new foundation.The principle of res judicata principally serves the credibility of the justice system in a given jurisdiction by prohibiting several tribunals, all acting within the parameters of their jurisdiction, from contradicting each other’s interpretation of the same facts. For this reason, the principle of res judicata does not provide an adequate basis for a transnationalised rule of ne bis in idem.From a human rights perspective, multiple prosecutions against the same person for the same facts collides with protecting individuals against arbitrary judicial treatment. This is true whether the multiple prosecutions all take place in one country or in several different countries. The rule of ne bis in idem could therefore be regarded as a manifestation of the (new ‘principle of personal legal certainty’.

  9. COOLC, Ne-213 Liquid Scintillation Detector Neutron Spectra Unfolding

    International Nuclear Information System (INIS)

    1971-01-01

    1 - Nature of physical problem solved: COOLC is designed to calculate a neutron energy spectrum from a pulse-height spectrum produced by a detector system using the liquid scintillator NE-213. 2 - Method of solution: The program estimates the counts which would be observed in an ideal detector system having a response which is specified by the user. The solution implicitly takes into account the non-negativity of the desired neutron spectrum. The solution is obtained by finding a nearly optimal combination of slices through the spectrometer response functions such that their sum approximates the response of a channel of the ideal analyzer, and then uses the coefficients so determined to obtain an estimate of the desired neutron spectrum. 3 - Restrictions on the complexity of the problem: There are none noted

  10. NE seeks to sell power directly to customers

    International Nuclear Information System (INIS)

    Anon.

    1993-01-01

    Nuclear Electric, the state-owned company that operates nuclear power stations in England and Wales, has applied to compete directly with privatized electricity generating companies in the sale of electricity to major customers. Since its formation in 1990, NE has had to sell all of its electrical output through the so-called pool operated by the National Grid Company, and then to 12 regional distribution companies that have franchises for about 75 percent of electricity consumption in their regions. On the other hand, the two large companies that took over the fossil-fuel power stations at the time of privatization, and other new independent companies that are building combined-cycle gas-turbine plants, are allowed to conclude supply contracts directly with large industrial customers

  11. Calibration of NE213 detector in neutron measurement

    International Nuclear Information System (INIS)

    Akkurt, I

    2004-01-01

    Organic scintillator is one of the widely used materials in neutron measurement as they have good timing properties and a high hydrogen content. Calibration of the detector system is an important part of the experimental study for interpretation of the results. As the neutron uncharged, the pulse from the detector is not directly used to determine neutron kinetic energy but the detection threshold for recoil charged particles (p,d,□ etc) has to be known in order to calculate the neutron detection efficiency. In this work calibration procedure of a NE213 detector array used in neutron measurements at MaxLab (Lund,Sweden ), is described.This includes both pulse height and neutron flight time which is important in neutron energy determinations

  12. Restaurant closure for the Jeûne genevois

    CERN Multimedia

    2007-01-01

    Restaurant 1 will be closed on Thursday 6th September (Jeûne Genevois) as well as Friday 7th, Saturday 8th and Sunday 9th September for technical reasons. During this time, Restaurant 2 will be open at the following times: –\tThursday 6th September: 9:00 – 20:00 –\tFriday 7th September: 8:00 – 20:00 –\tSaturday 8th and Sunday 9th September: 9:00–20:00 Hot meals will be served on all 4 days from 12:00 to 14:00 and from 18:00 to 19:30. For more information please see http://cern.ch/restaurant2 Thank you for your understanding.

  13. La chaîne du froid en agroalimentaire

    OpenAIRE

    Rosset , Philippe; Beaufort , Annie; Cornu , Marie; Poumeyrol , Gérard

    2002-01-01

    Le recours au froid constitue une pratique courante pour assurer une conservation prolongée des aliments, de quelques jours à quelques semaines. Limitant notre propos aux denrées réfrigérées et au risque sanitaire d'origine microbiologique, après un rappel de la définition de la chaîne du froid et des modalités générales de mise en oeuvre, nous aborderons dans un premier temps les particularités technologiques de son application. Celle-ci sera étudiée tout d'abord selon le type d'aliments con...

  14. Daily extreme temperature multifractals in Catalonia (NE Spain)

    Energy Technology Data Exchange (ETDEWEB)

    Burgueño, A. [Departament d' Astronomia i Meteorologia, Universitat de Barcelona, Barcelona (Spain); Lana, X., E-mail: francisco.javier.lana@upc.edu [Departament de Física i Enginyeria Nuclear, Universitat Politècnica de Catalunya, Barcelona (Spain); Serra, C. [Departament de Física i Enginyeria Nuclear, Universitat Politècnica de Catalunya, Barcelona (Spain); Martínez, M.D. [Departament de Física Aplicada, Universitat Politècnica de Catalunya, Barcelona (Spain)

    2014-02-01

    The multifractal character of the daily extreme temperatures in Catalonia (NE Spain) is analyzed by means of the multifractal detrended fluctuation analysis (MF-DFA) applied to 65 thermometric records covering years 1950–2004. Although no clear spatial patterns of the multifractal spectrum parameters appear, factor scores deduced from Principal Component analysis indicate some signs of spatial gradients. Additionally, the daily extreme temperature series are classified depending on their complex time behavior, through four multifractal parameters (Hurst exponent, Hölder exponent with maximum spectrum, spectrum asymmetry and spectrum width). As a synthesis of the three last parameters, a basic measure of complexity is proposed through a normalized Complexity Index. Its regional behavior is found to be free of geographical dependences. This index represents a new step towards the description of the daily extreme temperatures complexity.

  15. First Measurement of Neutrino Interactions in MicroBooNE

    Energy Technology Data Exchange (ETDEWEB)

    Hamilton, Pip [Fermilab

    2016-11-02

    The MicroBooNE detector has recently completed its first year of neutrino beam data-taking in the Booster Neutrino Beam at Fermilab, having collected approximately half of its intended data ($3.4\\times10^{20}$ of $6.6\\times10^{20}$ protons on target). We present kinematic distributions of neutrino interactions observed from a small subset of this data (equivalent to $5\\times10^{19}$ protons on target), both as a first step towards a charged-current muon neutrino cross-section on argon, and as an exploration of the capabilities and operational challenges of large liquid argon time projection chambers as neutrino detectors. These distributions have been assessed using fully automated event selection and reconstruction.

  16. α cluster structures in unbound states in 19Ne

    Science.gov (United States)

    Otani, Reiji; Iwasaki, Masataka; Ito, Makoto

    2016-06-01

    Cluster structures in 19Ne are studied by the microscopic and macroscopic cluster models. In the microscopic calculation, the coupled-channels problem of (3He+16O) + (α+15O) is solved, and the adiabatic energy surfaces, which are the series of the energy eigenvalues as a function of the He-O distance, are investigated. In the adiabatic energy curves, the several local minima are generated in the spatial region of the small core distance, where the neutron hole inside of the He or O nucleus is strongly coupled to the residual nuclei. The energy spectra, which are constructed from the strong coupling states, nicely reproduce the the low-lying energy levels in the 19Ne nucleus. In the macroscopic approach, the α + 15O potential is evaluated from the elastic scattering of the α + 15N system, and the resonant levels of the α + 15O system are calculated under the absorbing boundary condition. The potential model predicts the existence of the resonances above the α threshold, which has a weak-coupling scheme of the α particle and one hole inside of the 16O nucleus. The extended microscopic calculations of (3He+16O) + (α+15O) + (5He+14O) are performed in order to see the coupling effect of the 5p-2h configuration, which corresponds to the shell model limit of the 5He + 14O cluster configuration. The extended calculation suggests that the 5He + 14O configuration plays an important role on the formation of the 3/2+ resonance at 0.5 MeV with respect to the α threshold.

  17. α cluster structures in unbound states in 19Ne

    Directory of Open Access Journals (Sweden)

    Otani Reiji

    2016-01-01

    Full Text Available Cluster structures in 19Ne are studied by the microscopic and macroscopic cluster models. In the microscopic calculation, the coupled-channels problem of (3He+16O + (α+15O is solved, and the adiabatic energy surfaces, which are the series of the energy eigenvalues as a function of the He–O distance, are investigated. In the adiabatic energy curves, the several local minima are generated in the spatial region of the small core distance, where the neutron hole inside of the He or O nucleus is strongly coupled to the residual nuclei. The energy spectra, which are constructed from the strong coupling states, nicely reproduce the the low-lying energy levels in the 19Ne nucleus. In the macroscopic approach, the α + 15O potential is evaluated from the elastic scattering of the α + 15N system, and the resonant levels of the α + 15O system are calculated under the absorbing boundary condition. The potential model predicts the existence of the resonances above the α threshold, which has a weak-coupling scheme of the α particle and one hole inside of the 16O nucleus. The extended microscopic calculations of (3He+16O + (α+15O + (5He+14O are performed in order to see the coupling effect of the 5p-2h configuration, which corresponds to the shell model limit of the 5He + 14O cluster configuration. The extended calculation suggests that the 5He + 14O configuration plays an important role on the formation of the 3/2+ resonance at 0.5 MeV with respect to the α threshold.

  18. Measurement of 17F(d ,n )18Ne and the impact on the 17F(p ,γ )18Ne reaction rate for astrophysics

    Science.gov (United States)

    Kuvin, S. A.; Belarge, J.; Baby, L. T.; Baker, J.; Wiedenhöver, I.; Höflich, P.; Volya, A.; Blackmon, J. C.; Deibel, C. M.; Gardiner, H. E.; Lai, J.; Linhardt, L. E.; Macon, K. T.; Rasco, B. C.; Quails, N.; Colbert, K.; Gay, D. L.; Keeley, N.

    2017-10-01

    Background: The 17F(p ,γ )18Ne reaction is part of the astrophysical "hot CNO" cycles that are important in astrophysical environments like novas. Its thermal reaction rate is low owing to the relatively high energy of the resonances and therefore is dominated by direct, nonresonant capture in stellar environments at temperatures below 0.4 GK. Purpose: An experimental method is established to extract the proton strength to bound and unbound states in experiments with radioactive ion beams and to determine the parameters of direct and resonant capture in the 17F(p ,γ )18Ne reaction. Method: The 17F(d ,n )18Ne reaction is measured in inverse kinematics using a beam of the short-lived isotope 17F and a compact setup of neutron, proton, γ -ray, and heavy-ion detectors called resoneut. Results: The spectroscopic factors for the lowest l =0 proton resonances at Ec .m .=0.60 and 1.17 MeV are determined, yielding results consistent within 1.4 σ of previous proton elastic-scattering measurements. The asymptotic normalization coefficients of the bound 21+ and 22+ states in 18Ne are determined and the resulting direct-capture reaction rates are extracted. Conclusions: The direct-capture component of the 17F(p ,γ )18Ne reaction is determined for the first time from experimental data on 18Ne.

  19. 77 FR 4713 - Proposed Establishment of Class E Airspace; Red Cloud, NE

    Science.gov (United States)

    2012-01-31

    ...-0426; Airspace Docket No. 11-ACE-7] Proposed Establishment of Class E Airspace; Red Cloud, NE AGENCY... action proposes to establish Class E airspace at Red Cloud, NE. Controlled airspace is necessary to accommodate new Standard Instrument Approach Procedures (SIAP) at Red Cloud Municipal Airport. The FAA is...

  20. 77 FR 29871 - Establishment of Class E Airspace; Red Cloud, NE

    Science.gov (United States)

    2012-05-21

    ...-0426; Airspace Docket No. 11-ACE-7] Establishment of Class E Airspace; Red Cloud, NE AGENCY: Federal... at Red Cloud, NE. Controlled airspace is necessary to accommodate new Area Navigation (RNAV) Standard Instrument Approach Procedures at Red Cloud Municipal Airport. The FAA is taking this action to enhance the...

  1. Ne bis in idem põhimõte Euroopa Liidu õiguses / Uno Lõhmus

    Index Scriptorium Estoniae

    Lõhmus, Uno, 1952-

    2009-01-01

    Ne bis in idem põhimõtte ehk teistkordse kohtumõistmise ja karistamise keelu territoriaalsest kohaldamisest. Schengeni rakenduskonventsiooni artiklis 54 sisalduva ne bis in idem põhimõtte tõlgendustest. Mõistetest "sama tegu" ja "lõplik kohtuotsus"

  2. Desktop Video: Multi-Media on the NeXT Computer.

    Science.gov (United States)

    Stammen, Ronald M.; Richardson, Jolene

    A new course, Independent Study Research and Writing via Telecommunications, is being developed by the Division of Independent Study (DIS) of the North Dakota Department of Public Instruction to teach telepublishing skills utilizing the NeXT telecommunicating (interpersonal computing) techniques, i.e., NeXT Mail. This multimedia electronic-mail…

  3. Using ICD for structural analysis of clusters: a case study on NeAr clusters

    Science.gov (United States)

    Fasshauer, E.; Förstel, M.; Pallmann, S.; Pernpointner, M.; Hergenhahn, U.

    2014-10-01

    We present a method to utilize interatomic Coulombic decay (ICD) to retrieve information about the mean geometric structures of heteronuclear clusters. It is based on observation and modelling of competing ICD channels, which involve the same initial vacancy, but energetically different final states with vacancies in different components of the cluster. Using binary rare gas clusters of Ne and Ar as an example, we measure the relative intensity of ICD into (Ne+)2 and Ne+Ar+ final states with spectroscopically well separated ICD peaks. We compare in detail the experimental ratios of the Ne-Ne and Ne-Ar ICD contributions and their positions and widths to values calculated for a diverse set of possible structures. We conclude that NeAr clusters exhibit a core-shell structure with an argon core surrounded by complete neon shells and, possibly, further an incomplete shell of neon atoms for the experimental conditions investigated. Our analysis allows one to differentiate between clusters of similar size and stochiometric Ar content, but different internal structure. We find evidence for ICD of Ne 2s-1, producing Ar+ vacancies in the second coordination shell of the initial site.

  4. 1 1) Pourquoi est-ce que je ne parviens pas à ouvrir les formulaires ...

    International Development Research Centre (IDRC) Digital Library (Canada)

    IDRC CRDI

    Faites vos calculs avec soin car votre demande pourrait ne pas être admissible.) 6) Existe-t-il une liste de pays admissibles dans lesquels mon établissement ou organisme est autorisé à travailler ou à avoir des partenaires ? Non, le CRDI n'a rien de tel; cependant, il ne soutient des travaux que dans les pays à faible.

  5. The neXtProt knowledgebase on human proteins: 2017 update.

    Science.gov (United States)

    Gaudet, Pascale; Michel, Pierre-André; Zahn-Zabal, Monique; Britan, Aurore; Cusin, Isabelle; Domagalski, Marcin; Duek, Paula D; Gateau, Alain; Gleizes, Anne; Hinard, Valérie; Rech de Laval, Valentine; Lin, JinJin; Nikitin, Frederic; Schaeffer, Mathieu; Teixeira, Daniel; Lane, Lydie; Bairoch, Amos

    2017-01-04

    The neXtProt human protein knowledgebase (https://www.nextprot.org) continues to add new content and tools, with a focus on proteomics and genetic variation data. neXtProt now has proteomics data for over 85% of the human proteins, as well as new tools tailored to the proteomics community.Moreover, the neXtProt release 2016-08-25 includes over 8000 phenotypic observations for over 4000 variations in a number of genes involved in hereditary cancers and channelopathies. These changes are presented in the current neXtProt update. All of the neXtProt data are available via our user interface and FTP site. We also provide an API access and a SPARQL endpoint for more technical applications. © The Author(s) 2016. Published by Oxford University Press on behalf of Nucleic Acids Research.

  6. Bůh Tě vyzdvihl nad ostatní, a proto musíš být moudřejší než oni: obraz Karla IV. jako mudrce na trůně

    Czech Academy of Sciences Publication Activity Database

    Žůrek, Václav

    2016-01-01

    Roč. 38, č. 4 (2016), s. 18-21 ISSN 0418-5129 Grant - others:ERC(CZ) Origins of the Vernacular Mode. Regional Identities and European Networks in Late Medieval Europe Institutional support: RVO:67985955 Keywords : Mirror for princes * Charles IV * Charles university * Heinrich von Mügeln * Salomon Subject RIV: AB - History

  7. Addition of exogenous NAD+ prevents mefloquine-induced neuroaxonal and hair cell degeneration through reduction of caspase-3-mediated apoptosis in cochlear organotypic cultures.

    Directory of Open Access Journals (Sweden)

    Dalian Ding

    Full Text Available Mefloquine is widely used for the treatment of malaria. However, this drug is known to induce neurological side effects including depression, anxiety, balance disorder, and sensorineural hearing loss. Yet, there is currently no treatment for these side effects.In this study, we show that the coenzyme NAD(+, known to play a critical role in maintaining the appropriate cellular redox environment, protects cochlear axons and sensory hair cells from mefloquine-induced degeneration in cultured rat cochleae. Mefloquine alone destroyed hair cells and nerve fiber axons in rat cochlear organotypics cultures in a dose-dependent manner, while treatment with NAD(+ protected axons and hair cells from mefloquine-induced degeneration. Furthermore, cochlear organs treated with mefloquine showed increased oxidative stress marker levels, including superoxide and protein carbonyl, and increased apoptosis marker levels, including TUNEL-positive nuclei and caspases-3. Treatment with NAD(+ reduced the levels of these oxidative stress and apoptosis markers.Taken together, our findings suggest that that mefloquine disrupts the cellular redox environment and induces oxidative stress in cochlear hair cells and nerve fibers leading to caspases-3-mediated apoptosis of these structures. Exogenous NAD(+ suppresses mefloquine-induced oxidative stress and prevents the degeneration of cochlear axons and sensory hair cells caused by mefloquine treatment.

  8. Addition of exogenous NAD+ prevents mefloquine-induced neuroaxonal and hair cell degeneration through reduction of caspase-3-mediated apoptosis in cochlear organotypic cultures.

    Science.gov (United States)

    Ding, Dalian; Qi, Weidong; Yu, Dongzhen; Jiang, Haiyan; Han, Chul; Kim, Mi-Jung; Katsuno, Kana; Hsieh, Yun Hua; Miyakawa, Takuya; Salvi, Richard; Tanokura, Masaru; Someya, Shinichi

    2013-01-01

    Mefloquine is widely used for the treatment of malaria. However, this drug is known to induce neurological side effects including depression, anxiety, balance disorder, and sensorineural hearing loss. Yet, there is currently no treatment for these side effects. In this study, we show that the coenzyme NAD(+), known to play a critical role in maintaining the appropriate cellular redox environment, protects cochlear axons and sensory hair cells from mefloquine-induced degeneration in cultured rat cochleae. Mefloquine alone destroyed hair cells and nerve fiber axons in rat cochlear organotypics cultures in a dose-dependent manner, while treatment with NAD(+) protected axons and hair cells from mefloquine-induced degeneration. Furthermore, cochlear organs treated with mefloquine showed increased oxidative stress marker levels, including superoxide and protein carbonyl, and increased apoptosis marker levels, including TUNEL-positive nuclei and caspases-3. Treatment with NAD(+) reduced the levels of these oxidative stress and apoptosis markers. Taken together, our findings suggest that that mefloquine disrupts the cellular redox environment and induces oxidative stress in cochlear hair cells and nerve fibers leading to caspases-3-mediated apoptosis of these structures. Exogenous NAD(+) suppresses mefloquine-induced oxidative stress and prevents the degeneration of cochlear axons and sensory hair cells caused by mefloquine treatment.

  9. "Õhtuti süütasid nad kassi haual küünla..." : [luuletused] / Tomi Kontio ; soome keelest tõlkinud Jan Kaus

    Index Scriptorium Estoniae

    Kontio, Tomi

    2007-01-01

    Sisu: "Õhtuti süütasid nad kassi haual küünla..." ; "Meri on plekkis, terasest neiu seisab..." ; "Lumi puude okstel, kirmetis aknal, jää..." ; Kontula kaar ; Vastavalt tahtele ; Ühel kaardil ; Esimene lumi ; Dimensioonid ; Pieta

  10. In vivo relevance of two critical levels for NAD(P)H:quinone oxidoreductase (NQO1)-mediated cellular protection against electrophile toxicity found in vitro

    NARCIS (Netherlands)

    Haan, de L.H.J.; Pot, G.K.; Aarts, J.M.M.J.G.; Rietjens, I.M.C.M.; Alink, G.M.

    2006-01-01

    NAD(P)H:quinone oxidoreductase (NQO1)-mediated detoxification of quinones is suggested to be involved in cancer prevention. In the present study, using transfected CHO cells, it was demonstrated that the relation between NQO1 activity and the resulting protection against the cytotoxicity of

  11. In vivo relevance of two critical levels for NAD(P)H:quinone oxidoreductase (NQO1)-mediated cellular protection against electrophile toxicity found in vitro.

    NARCIS (Netherlands)

    de Haan, L.H.; Pot, G.K.; Aarts, J.M.; Rietjens, I.M.; Alink, G.M.

    2006-01-01

    NAD(P)H:quinone oxidoreductase (NQO1)-mediated detoxification of quinones is suggested to be involved in cancer prevention. In the present study, using transfected CHO cells, it was demonstrated that the relation between NQO1 activity and the resulting protection against the cytotoxicity of

  12. Human NAD(P)H:quinone oxidoreductase 1 (NQO1)-mediated inactivation of reactive quinoneimine metabolites of diclofenac and mefenamic acid

    NARCIS (Netherlands)

    Vredenburg, Galvin; Elias, Naura S; Venkataraman, Harini; Hendriks, Delilah F G; Vermeulen, Nico P E; Commandeur, Jan N M; Vos, J Chris

    2014-01-01

    NAD(P)H: quinone oxidoreductase 1 (NQO1) is an enzyme capable of reducing a broad range of chemically reactive quinones and quinoneimines (QIs) and can be strongly upregulated by Nrf2/Keap1-mediated stress responses. Several commonly used drugs implicated in adverse drug reactions (ADRs) are known

  13. Reduction and Scavenging of Chemically Reactive Drug Metabolites by NAD(P)H:Quinone Oxidoreductase 1 and NRH:Quinone Oxidoreductase 2 and Variability in Hepatic Concentrations

    NARCIS (Netherlands)

    den Braver-Sewradj, Shalenie P; den Braver, Michiel W; Toorneman, Robin M; van Leeuwen, Stephanie; Zhang, Yongjie; Dekker, Stefan J; Vermeulen, Nico P E; Commandeur, Jan N M; Vos, J Chris

    2018-01-01

    Detoxicating enzymes NAD(P)H:quinone oxidoreductase 1 (NQO1) and NRH:quinone oxidoreductase 2 (NQO2) catalyze the two-electron reduction of quinone-like compounds. The protective role of the polymorphic NQO1 and NQO2 enzymes is especially of interest in the liver as the major site of drug

  14. Purification and characterization of NAD(P)H quinone reductase from the latex of Hevea brasiliensis Müll.-Arg. (Euphorbiaceae).

    Science.gov (United States)

    Chareonthiphakorn, Nopphakaew; Wititsuwannakul, Dhirayos; Golan-Goldhirsh, Avi; Wititsuwannakul, Rapepun

    2002-09-01

    NAD(P)H quinone reductase [NAD(P)H-QR] present in the latex of Hevea brasiliensis Müll.-Arg. (Euphorbiaceae) was purified to homogeniety from the B-serum fraction obtained by freeze-thawing of the bottom fraction of ultracentrifuged fresh latex. The purification protocol involved acetone fractionation, heat treatment, ion exchange chromatography and affinity chromatography. The M(r) determined by SDS-PAGE for the protein subunit was 21 kDa, and the molecular mass of the native enzyme estimated by gel filtration was 83 kDa, indicating that the native enzyme is a homotetramer. The enzyme showed pH stability over a range of 6 to at least 10 (with an optimum at pH 8) and thermal stability up to 80 degrees C. High NAD(P)H-QR activity (70%) was still retained after 10 h of preincubation at 80 degrees C. A comparable substrate specificity for this enzyme was observed among menadione, p-benzoquinone, juglone, and plumbagin, with only duroquinone generating a lower activity. Positive correlations between latex NAD(P)H-QR activity and rubber yield per tapping [fresh latex (r=0.89, PPP<0.01) indicated that enzyme activity could possibly be used as a marker to predict the yield potential of selected clones.

  15. Stereo-specificity for pro-(R) hydrogen of NAD(P)H during enzyme-catalyzed hydride transfer to CL-20

    International Nuclear Information System (INIS)

    Bhushan, Bharat; Halasz, Annamaria; Hawari, Jalal

    2005-01-01

    A dehydrogenase from Clostridium sp. EDB2 and a diaphorase from Clostridium kluyveri were reacted with CL-20 to gain insights into the enzyme-catalyzed hydride transfer to CL-20, and the enzyme's stereo-specificity for either pro-R or pro-S hydrogens of NAD(P)H. Both enzymes biotransformed CL-20 at rates of 18.5 and 24 nmol/h/mg protein, using NADH and NADPH as hydride-source, respectively, to produce a N-denitrohydrogenated product with a molecular weight of 393 Da. In enzyme kinetics studies using reduced deuterated pyridine nucleotides, we found a kinetic deuterium isotopic effect of 2-fold on CL-20 biotransformation rate using dehydrogenase enzyme against (R)NADD as a hydride-source compared to either (S)NADD or NADH. Whereas, in case of diaphorase, the kinetic deuterium isotopic effect of about 1.5-fold was observed on CL-20 biotransformation rate using (R)NADPD as hydride-source. In a comparative study with LC-MS, using deuterated and non-deuterated NAD(P)H, we found a positive mass-shift of 1 Da in the N-denitrohydrogenated product suggesting the involvement of a deuteride (D - ) transfer from NAD(P)D. The present study thus revealed that both dehydrogenase and diaphorase enzymes from the two Clostridium species catalyzed a hydride transfer to CL-20 and showed stereo-specificity for pro-R hydrogen of NAD(P)H

  16. Escherichia coli phnN, encoding ribose 1,5-bisphosphokinase activity (phosphoribosyl diphosphate forming): dual role in phosphonate degradation and NAD biosynthesis pathways

    DEFF Research Database (Denmark)

    Hove-Jensen, Bjarne; Rosenkrantz, Tina J; Haldimann, Andreas

    2003-01-01

    An enzymatic pathway for synthesis of 5-phospho-D-ribosyl alpha-1-diphosphate (PRPP) without the participation of PRPP synthase was analyzed in Escherichia coli. This pathway was revealed by selection for suppression of the NAD requirement of strains with a deletion of the prs gene, the gene...

  17. Search of fission products in 20Ne-ion beam interaction with 165Ho at 8 MeV/nucleon

    International Nuclear Information System (INIS)

    Singh, D.; Ali, R.; Afzal Ansari, M.; Rashid, M.H.

    2006-01-01

    In the present work, during the study complete fusion (CF) and incomplete fusion (ICF) in 20 Ne-induced reactions, the production cross-sections for several fission products in 20 Ne + 165 Ho system have been measured

  18. Ne bis in idem põhimõte Eesti karistusõiguses : [bakalaureusetöö] / Natalja Mogiljova ; Tartu Ülikool, õigusteaduskond ; juhendaja: Elina Elkind

    Index Scriptorium Estoniae

    Mogiljova, Natalja

    2009-01-01

    Ne bis in idem põhimõtte olemusest ja ajaloolisest arengust, horisontaalsest riigisisesest ne bis in idem-ist, rahvusvahelistest konventsioonidest, EIÕK praktikast, Riigikohtu praktikast, ne bis in idem Ameerika Ühendriikide õiguskorras

  19. Interaction of Ne(2p54p), Ar(3p54p) and Kr(4p55p) excited atoms with He and Ne atoms. Processes of collisional depolarization

    International Nuclear Information System (INIS)

    Zagrebin, A.L.; Lednev, M.G.

    1990-01-01

    Quasimolecular terms Ne(2p 5 4p)+He, Ar(3p 5 4p)+He,Ne and Kr(4p 5 5p)+He,Ne are calculated within the framework of one-configuration method of effective Hamiltonian. The results of calculations agree with the experimental data

  20. The MiniBooNE detector technical design report

    Energy Technology Data Exchange (ETDEWEB)

    I. Stancu et al.

    2003-04-18

    The MiniBooNE experiment [1] is motivated by the LSND observation, [2] which has been interpreted as {nu}{sub {mu}} {yields} {nu}{sub e} oscillations, and by the atmospheric neutrino deficit, [3,4,5] which may be ascribed to {nu}{sub {mu}} oscillations into another type of neutrino. MiniBooNE is a single-detector experiment designed to: obtain {approx} 1000 {nu}{sub {mu}} {yields} {nu}{sub e} events if the LSND signal is due to {nu}{sub {mu}} {yields} {nu}{sub e} oscillations, establishing the oscillation signal at the > 5{sigma} level as shown in Fig. 1.1; extend the search for {nu}{sub {mu}} {yields} {nu}{sub e} oscillations significantly beyond what has been studied previously if no signal is observed; search for {nu}{sub {mu}} disappearance to address the atmospheric neutrino deficit with a signal that is a suppression of the rate of {nu}{sub {mu}}C {yields} {mu}N events from the expected 600,000 per year; measure the oscillation parameters as shown in Fig. 1.2 if oscillations are observed; and test CP conservation in the lepton sector if oscillations are observed by running with separate {nu}{sub {mu}} and {bar {nu}}{sub {mu}} beams. The detector will consist of a spherical tank 6.1 m (20 feet) in radius, as shown in Fig. 1.3, that stands in a 45-foot diameter cylindrical vault. An inner tank structure at 5.75 m radius will support 1280 8-inch phototubes (10% coverage) pointed inward and optically isolated from the outer region of the tank. The tank will be filled with 807 t of mineral oil, resulting in a 445 t fiducial volume. The outer tank volume will serve as a veto shield for identifying particles both entering and leaving the detector with 240 phototubes mounted on the tank wall. Above the detector tank will be an electronics enclosure that houses the fast electronics and data acquisition system and a utilities enclosure that houses the plumbing, overflow tank, and calibration laser. The detector will be located {approx} 550 m from the Booster neutrino

  1. Microseismicity and seismotectonics of the South Caspian Lowlands, NE Iran

    Science.gov (United States)

    Nemati, Majid; Hollingsworth, James; Zhan, Zhongwen; Bolourchi, Mohammad Javad; Talebian, Morteza

    2013-06-01

    This paper is concerned with the microseismicity and seismotectonics of the eastern South Caspian Sea region, where the East Alborz mountains descend to meet the South Caspian Lowlands of NE Iran. To better understand the present-day tectonics and seismicity of this region, which includes the cities of Gorgan and Gonbad-e-Qabus (combined population 500 000), we installed a temporary local seismic network across the area for 6 months between 2009 and 2010. We analysed the seismicity and focal mechanisms together with data from the permanent networks of the Institute of Geophysics, University of Tehran (IGUT) and the International Institute of Earthquake Engineering and Seismology (IIEES), based in Tehran. Microseismicity is focused primarily on the Shahrud fault system, which bounds the east Alborz range to the south. Relatively few earthquakes are associated with the Khazar thrust fault, which bounds the north side of the range. A cluster of shallow microseismicity (Khazar fault (within the South Caspian Lowlands; SCL), an area typically thought to be non-deforming. This area coincides with the location of three relatively deep thrust earthquakes (Mw 5.3-5.5) which occurred in 1999, 2004 and 2005. Inversion of teleseismic body waveforms allows us to constrain the depth of these earthquakes at 26-29 km. Although significant sedimentation throughout the SCL obscures any expression of recent fault activity at the surface, focal mechanisms of well-located events from the shallow cluster of micro-seismicity show a significant component of left-lateral strike-slip motion (assuming slip occurs on NE-SW fault planes, typical of active faults in the region), as well as a small normal component. Inversion of traveltimes for well-located events in our network yields a velocity structure for the region, and a Moho depth of 41 km. The pattern of deep thrust and shallow normal seismicity could be explained by bending of the rigid South Caspian crust as it underthrusts the East

  2. Nicotinamide riboside, a form of vitamin B3 and NAD+ precursor, relieves the nociceptive and aversive dimensions of paclitaxel-induced peripheral neuropathy in female rats.

    Science.gov (United States)

    Hamity, Marta V; White, Stephanie R; Walder, Roxanne Y; Schmidt, Mark S; Brenner, Charles; Hammond, Donna L

    2017-05-01

    Injury to sensory afferents may contribute to the peripheral neuropathies that develop after administration of chemotherapeutic agents. Manipulations that increase levels of nicotinamide adenine dinucleotide (NAD) can protect against neuronal injury. This study examined whether nicotinamide riboside (NR), a third form of vitamin B3 and precursor of NAD, diminishes tactile hypersensitivity and place escape-avoidance behaviors in a rodent model of paclitaxel-induced peripheral neuropathy. Female Sprague-Dawley rats received 3 intravenous injections of 6.6 mg/kg paclitaxel over 5 days. Daily oral administration of 200 mg/kg NR beginning 7 days before paclitaxel treatment and continuing for another 24 days prevented the development of tactile hypersensitivity and blunted place escape-avoidance behaviors. These effects were sustained after a 2-week washout period. This dose of NR increased blood levels of NAD by 50%, did not interfere with the myelosuppressive effects of paclitaxel, and did not produce adverse locomotor effects. Treatment with 200 mg/kg NR for 3 weeks after paclitaxel reversed the well-established tactile hypersensitivity in a subset of rats and blunted escape-avoidance behaviors. Pretreatment with 100 mg/kg oral acetyl-L-carnitine (ALCAR) did not prevent paclitaxel-induced tactile hypersensitivity or blunt escape-avoidance behaviors. ALCAR by itself produced tactile hypersensitivity. These findings suggest that agents that increase NAD, a critical cofactor for mitochondrial oxidative phosphorylation systems and cellular redox systems involved with fuel utilization and energy metabolism, represent a novel therapeutic approach for relief of chemotherapy-induced peripheral neuropathies. Because NR is a vitamin B3 precursor of NAD and a nutritional supplement, clinical tests of this hypothesis may be accelerated.

  3. Head to head comparison of short-term treatment with the NAD+ precursor nicotinamide mononucleotide (NMN and six weeks of exercise in obese female mice

    Directory of Open Access Journals (Sweden)

    Golam Mezbah Uddin

    2016-08-01

    Full Text Available Obesity is well known to be a major cause of several chronic metabolic diseases, which can be partially counteracted by exercise. This is due, in part, to an upregulation of mitochondrial activity through increased nicotinamide adenine dinucleotide (NAD+. Recent studies have shown that NAD+ levels can be increased by using the NAD+ precursor, nicotinamide mononucleotide (NMN leading to the suggestion that NMN could be a useful intervention in diet related metabolic disorders. In this study we compared the metabolic, and especially mitochondrial-associated, effects of exercise and NMN in ameliorating the consequences of high-fat diet (HFD induced obesity in mice. Sixty female 5 week old C57BL6/J mice were allocated across 5 interventions: Chow sedentary: CS; Chow exercise: CEX; HFD sedentary: HS; HFD NMN: HNMN; HFD exercise: HEX (12/group. After 6 weeks of diet, exercise groups underwent treadmill exercise (15 m/min for 45 minutes, 6 days per week for 6 weeks. NMN or vehicle (500 mg/kg body weight was injected (i.p. daily for the last 17 days. No significant alteration in body weight was observed in response to exercise or NMN. The HFD significantly altered adiposity, glucose tolerance, plasma insulin, NADH levels and citrate synthase activity in muscle and liver. HEX and HNMN groups both showed significantly improved glucose tolerance compared to the HS group. NAD+ levels were increased significantly both in muscle and liver by NMN whereas exercise increased NAD+ only in muscle. Both NMN and exercise ameliorated the HFD-induced reduction in liver citrate synthase activity. However, exercise, but not NMN, ameliorated citrate synthase activity in muscle. Overall these data suggest that while exercise and NMN-supplementation can induce similar reversal of the glucose intolerance induced by obesity, they are associated with tissue-specific effects and differential alterations to mitochondrial function in muscle and liver.

  4. The Nuclear Receptor LXR Limits Bacterial Infection of Host Macrophages through a Mechanism that Impacts Cellular NAD Metabolism

    Directory of Open Access Journals (Sweden)

    Jonathan Matalonga

    2017-01-01

    Full Text Available Macrophages exert potent effector functions against invading microorganisms but constitute, paradoxically, a preferential niche for many bacterial strains to replicate. Using a model of infection by Salmonella Typhimurium, we have identified a molecular mechanism regulated by the nuclear receptor LXR that limits infection of host macrophages through transcriptional activation of the multifunctional enzyme CD38. LXR agonists reduced the intracellular levels of NAD+ in a CD38-dependent manner, counteracting pathogen-induced changes in macrophage morphology and the distribution of the F-actin cytoskeleton and reducing the capability of non-opsonized Salmonella to infect macrophages. Remarkably, pharmacological treatment with an LXR agonist ameliorated clinical signs associated with Salmonella infection in vivo, and these effects were dependent on CD38 expression in bone-marrow-derived cells. Altogether, this work reveals an unappreciated role for CD38 in bacterial-host cell interaction that can be pharmacologically exploited by activation of the LXR pathway.

  5. NAD(P-DEPENDENT DEHYDROGENASE ACTIVITY IN PERIPHERAL BLOOD LYMPHOCYTES OF INFANTS WITH ENLARGEMENT OF PHARYNGEAL TONSILS

    Directory of Open Access Journals (Sweden)

    L. M. Kurtasova

    2014-01-01

    Full Text Available We have observed and examined 57 children 1 to 3 years old diagnosed with enlargement of pharyngeal tonsils. A control group was presented by 35 healthy children. Bioluminescence technique was applied for studying NAD(P-dependent dehydrogenase activity in peripheral blood lymphocytes. Activation of aerobic respiration and increasing activity of pentose phosphate cycle-dependent plastic processes were registered in blood lymphocytes of children with hypertrophic pharyngeal tonsils; along with decreased function of malate-aspartate shunt in energy metabolism of the cells, diminished anaerobic reaction of NADHdependent LDH, lower interaction between Krebs cycle and reactions of amino acid metabolism, and reduced activity of glutathione reductase.

  6. Effects of Al on the splenic immune function and NE in rats.

    Science.gov (United States)

    Hu, Chongwei; Li, Jing; Zhu, Yanzhu; Bai, Chongsheng; Zhang, Jihong; Xia, Shiliang; Li, Yanfei

    2013-12-01

    Norepinephrine (NE) regulates the splenic immune function and it may be related to the effects of Aluminum (Al) on the splenic immune function. Here, the aim of this study was to further explore the effects of aluminum trichloride (AlCl3) on the splenic immune function and its relationship with NE. Forty male Wistar rats were orally exposed to AlCl3 (0, 64.18, 128.36 and 256.72 mg/kg BW) through drinking water for 120 days. The CD3(+), CD4(+), CD8(+) T lymphocytes, the T and B lymphocytes proliferation rates and serum NE concentration were examined. The correlation analysis between splenic immune function and NE were done. The results showed that the CD3(+), CD4(+), CD8(+) T lymphocytes and the T and B lymphocytes proliferation rates decreased and NE concentration increased in AlCl3-treated rats. NE was negatively correlated with proportions of CD3(+), CD4(+) T lymphocytes and T and B lymphocytes proliferation rates, but not correlated with CD8(+) T lymphocytes. The results suggest that AlCl3 suppresses the splenic immune function and NE plays important role in this process. Copyright © 2013 Elsevier Ltd. All rights reserved.

  7. Arabidopsis mTERF15 is required for mitochondrial nad2 intron 3 splicing and functional complex I activity.

    Directory of Open Access Journals (Sweden)

    Ya-Wen Hsu

    Full Text Available Mitochondria play a pivotal role in most eukaryotic cells, as they are responsible for the generation of energy and diverse metabolic intermediates for many cellular events. During endosymbiosis, approximately 99% of the genes encoded by the mitochondrial genome were transferred into the host nucleus, and mitochondria import more than 1000 nuclear-encoded proteins from the cytosol to maintain structural integrity and fundamental functions, including DNA replication, mRNA transcription and RNA metabolism of dozens of mitochondrial genes. In metazoans, a family of nuclear-encoded proteins called the mitochondrial transcription termination factors (mTERFs regulates mitochondrial transcription, including transcriptional termination and initiation, via their DNA-binding activities, and the dysfunction of individual mTERF members causes severe developmental defects. Arabidopsis thaliana and Oryza sativa contain 35 and 48 mTERFs, respectively, but the biological functions of only a few of these proteins have been explored. Here, we investigated the biological role and molecular mechanism of Arabidopsis mTERF15 in plant organelle metabolism using molecular genetics, cytological and biochemical approaches. The null homozygous T-DNA mutant of mTERF15, mterf15, was found to result in substantial retardation of both vegetative and reproductive development, which was fully complemented by the wild-type genomic sequence. Surprisingly, mitochondria-localized mTERF15 lacks obvious DNA-binding activity but processes mitochondrial nad2 intron 3 splicing through its RNA-binding ability. Impairment of this splicing event not only disrupted mitochondrial structure but also abolished the activity of mitochondrial respiratory chain complex I. These effects are in agreement with the severe phenotype of the mterf15 homozygous mutant. Our study suggests that Arabidopsis mTERF15 functions as a splicing factor for nad2 intron 3 splicing in mitochondria, which is essential

  8. Glycerol and environmental factors: effects on 1,3-propanediol production and NAD(+) regeneration in Lactobacillus panis PM1.

    Science.gov (United States)

    Kang, T S; Korber, D R; Tanaka, T

    2013-10-01

    This study was conducted to understand the influences of fermentation factors in NADH recycling and mechanisms of 1,3-propanediol (1,3-PDO) production in Lactobacillus panis PM1. We conducted metabolite analyses, qRT-PCR of the glycerol reductive pathway [glycerol dehydratase (DhaB) and 1,3-PDO dehydrogenase (DhaT)] and DhaT activity assays at different pH, temperature and initial glycerol concentrations. The supplementation of 150 mmol l(-1) glycerol caused a shift in NADH flux from ethanol to 1,3-PDO production, whereas 300 mol l(-1) glycerol negatively affected the regeneration of NAD(+) via 1,3-PDO production. This retardation decreased transcription levels and specific activities of DhaT. The decreased DhaT activity eventually caused the shutdown of 1,3-PDO production. Temperature and pH did not significantly affect the specific activity of DhaT, whereas expression of genes for DhaB and DhaT was activated under acidic conditions. Moreover, fresh glucose addition after its depletion could not restart the glycerol reduction, but increased ethanol production. Those environmental factors affect 1,3-PDO production in different ways through changing the expression level of enzymes and shifting the NAD(+) regeneration pathways. Our findings elucidated a key element to optimize 1,3-PDO production by Lact. panis PM1, which potentially improves 1,3-PDO manufacturing efficiencies. © 2013 The Society for Applied Microbiology.

  9. New biotechnological perspectives of a NADH oxidase variant from Thermus thermophilus HB27 as NAD+-recycling enzyme

    Directory of Open Access Journals (Sweden)

    Rocha-Martín Javier

    2011-11-01

    Full Text Available Abstract Background The number of biotransformations that use nicotinamide recycling systems is exponentially growing. For this reason one of the current challenges in biocatalysis is to develop and optimize more simple and efficient cofactor recycling systems. One promising approach to regenerate NAD+ pools is the use of NADH-oxidases that reduce oxygen to hydrogen peroxide while oxidizing NADH to NAD+. This class of enzymes may be applied to asymmetric reduction of prochiral substrates in order to obtain enantiopure compounds. Results The NADH-oxidase (NOX presented here is a flavoenzyme which needs exogenous FAD or FMN to reach its maximum velocity. Interestingly, this enzyme is 6-fold hyperactivated by incubation at high temperatures (80°C under limiting concentrations of flavin cofactor, a change that remains stable even at low temperatures (37°C. The hyperactivated form presented a high specific activity (37.5 U/mg at low temperatures despite isolation from a thermophile source. Immobilization of NOX onto agarose activated with glyoxyl groups yielded the most stable enzyme preparation (6-fold more stable than the hyperactivated soluble enzyme. The immobilized derivative was able to be reactivated under physiological conditions after inactivation by high solvent concentrations. The inactivation/reactivation cycle could be repeated at least three times, recovering full NOX activity in all cases after the reactivation step. This immobilized catalyst is presented as a recycling partner for a thermophile alcohol dehydrogenase in order to perform the kinetic resolution secondary alcohols. Conclusion We have designed, developed and characterized a heterogeneous and robust biocatalyst which has been used as recycling partner in the kinetic resolution of rac-1-phenylethanol. The high stability along with its capability to be reactivated makes this biocatalyst highly re-useable for cofactor recycling in redox biotransformations.

  10. Atomic-resolution structures of horse liver alcohol dehydrogenase with NAD(+) and fluoroalcohols define strained Michaelis complexes.

    Science.gov (United States)

    Plapp, Bryce V; Ramaswamy, S

    2012-05-15

    Structures of horse liver alcohol dehydrogenase complexed with NAD(+) and unreactive substrate analogues, 2,2,2-trifluoroethanol or 2,3,4,5,6-pentafluorobenzyl alcohol, were determined at 100 K at 1.12 or 1.14 Å resolution, providing estimates of atomic positions with overall errors of ~0.02 Å, the geometry of ligand binding, descriptions of alternative conformations of amino acid residues and waters, and evidence of a strained nicotinamide ring. The four independent subunits from the two homodimeric structures differ only slightly in the peptide backbone conformation. Alternative conformations for amino acid side chains were identified for 50 of the 748 residues in each complex, and Leu-57 and Leu-116 adopt different conformations to accommodate the different alcohols at the active site. Each fluoroalcohol occupies one position, and the fluorines of the alcohols are well-resolved. These structures closely resemble the expected Michaelis complexes with the pro-R hydrogens of the methylene carbons of the alcohols directed toward the re face of C4N of the nicotinamide rings with a C-C distance of 3.40 Å. The oxygens of the alcohols are ligated to the catalytic zinc at a distance expected for a zinc alkoxide (1.96 Å) and participate in a low-barrier hydrogen bond (2.52 Å) with the hydroxyl group of Ser-48 in a proton relay system. As determined by X-ray refinement with no restraints on bond distances and planarity, the nicotinamide rings in the two complexes are slightly puckered (quasi-boat conformation, with torsion angles of 5.9° for C4N and 4.8° for N1N relative to the plane of the other atoms) and have bond distances that are somewhat different compared to those found for NAD(P)(+). It appears that the nicotinamide ring is strained toward the transition state on the path to alcohol oxidation.

  11. Mitochondrial NAD+-dependent malic enzyme from Anopheles stephensi: a possible novel target for malaria mosquito control

    Directory of Open Access Journals (Sweden)

    Pon Jennifer

    2011-10-01

    Full Text Available Abstract Background Anopheles stephensi mitochondrial malic enzyme (ME emerged as having a relevant role in the provision of pyruvate for the Krebs' cycle because inhibition of this enzyme results in the complete abrogation of oxygen uptake by mitochondria. Therefore, the identification of ME in mitochondria from immortalized A. stephensi (ASE cells and the investigation of the stereoselectivity of malate analogues are relevant in understanding the physiological role of ME in cells of this important malaria parasite vector and its potential as a possible novel target for insecticide development. Methods To characterize the mitochondrial ME from immortalized ASE cells (Mos. 43; ASE, mass spectrometry analyses of trypsin fragments of ME, genomic sequence analysis and biochemical assays were performed to identify the enzyme and evaluate its activity in terms of cofactor dependency and inhibitor preference. Results The encoding gene sequence and primary sequences of several peptides from mitochondrial ME were found to be highly homologous to the mitochondrial ME from Anopheles gambiae (98% and 59% homologous to the mitochondrial NADP+-dependent ME isoform from Homo sapiens. Measurements of ME activity in mosquito mitochondria isolated from ASE cells showed that (i Vmax with NAD+ was 3-fold higher than that with NADP+, (ii addition of Mg2+ or Mn2+ increased the Vmax by 9- to 21-fold, with Mn2+ 2.3-fold more effective than Mg2+, (iii succinate and fumarate increased the activity by 2- and 5-fold, respectively, at sub-saturating concentrations of malate, (iv among the analogs of L-malate tested as inhibitors of the NAD+-dependent ME catalyzed reaction, small (2- to 3-carbons organic diacids carrying a 2-hydroxyl/keto group behaved as the most potent inhibitors of ME activity (e.g., oxaloacetate, tartronic acid and oxalate. Conclusions The biochemical characterization of Anopheles stephensi ME is of critical relevance given its important role in

  12. Root phosphoenolpyruvate carboxylase and NAD-malic enzymes activity increase the ammonium-assimilating capacity in tomato.

    Science.gov (United States)

    Setién, Igor; Vega-Mas, Izargi; Celestino, Natalia; Calleja-Cervantes, María Eréndira; González-Murua, Carmen; Estavillo, José María; González-Moro, María Begoña

    2014-03-01

    Plant ammonium tolerance has been associated with the capacity to accumulate large amounts of ammonium in the root vacuoles, to maintain carbohydrate synthesis and especially with the capacity of maintaining high levels of inorganic nitrogen assimilation in the roots. The tricarboxylic acid cycle (TCA) is considered a cornerstone in nitrogen metabolism, since it provides carbon skeletons for nitrogen assimilation. The hypothesis of this work was that the induction of anaplerotic routes of phosphoenolpyruvate carboxylase (PEPC), malate dehydrogenase (MDH) and malic enzyme (NAD-ME) would enhance tolerance to ammonium nutrition. An experiment was established with tomato plants (Agora Hybrid F1) grown under different ammonium concentrations. Growth parameters, metabolite contents and enzymatic activities related to nitrogen and carbon metabolism were determined. Unlike other tomato cultivars, tomato Agora Hybrid F1 proved to be tolerant to ammonium nutrition. Ammonium was assimilated as a biochemical detoxification mechanism, thus leading to the accumulation of Gln and Asn as free amino acids in both leaves and roots as an innocuous and transitory store of nitrogen, in addition to protein synthesis. When the concentration of ammonium in the nutrient solution was high, the cyclic operation of the TCA cycle seemed to be interrupted and would operate in two interconnected branches to provide α-ketoglutarate for ammonium assimilation: one branch supported by malate accumulation and by the induction of anaplerotic PEPC and NAD-ME in roots and MDH in leaves, and the other branch supported by stored citrate in the precedent dark period. Copyright © 2013 Elsevier GmbH. All rights reserved.

  13. Ne2 encodes protein(s) and the altered RuBisCO could be the proteomics leader of hybrid necrosis in wheat (Triticum aestivum L.).

    Science.gov (United States)

    Pan, Si Rui; Pan, Xing Lai; Pan, Qian Ying; Shi, Yin Hong; Zhang, Li; Fan, Yun; Xue, Yan Rui

    2017-06-01

    Wheat hybrid necrosis is caused by the interaction of two dominant complementary genes, Ne1 and Ne2, located on chromosome arms 5BL and 2BS, respectively. The sequences of Ne1 or Ne2 have not yet been identified. It is also not known whether Ne1 and Ne2 are structural or regulatory genes. Understanding the proteomic pathways may provide a knowledge base for protecting or maximizing the photosynthesis capacity of wheat. Using DIGE and MALDITOF- TOF MS, the flag leaf protein patterns of the two unique F14 near-isogenic line siblings (NILs), the necrotic ShunMai 12Ah (Ne1Ne1Ne2Ne2) and the normal ShunMai 12Af (Ne1Ne1ne2ne2) were compared. Due to the presence or absence of Ne2, (i) three protein spots were expressed or disappeared, (ii) seven RuBisCO-related proteins were altered significantly, and (iii) 21 photosynthesis/glucose related proteins were changed significantly. Three hypotheses were deduced, (i) Ne1 may also encode protein(s), (ii) genetic maladjustment of RuBisCO could lead to early leaf death, and (iii) interactions between nuclear genes and chloroplast genes could determine photosynthetic traits. Our hypothetical model presents the RuBisCO pathway of hybrid necrosis in wheat and explains how Ne1 and Ne2 interact at molecular level.

  14. NeSSM: a Next-generation Sequencing Simulator for Metagenomics.

    Directory of Open Access Journals (Sweden)

    Ben Jia

    Full Text Available BACKGROUND: Metagenomics can reveal the vast majority of microbes that have been missed by traditional cultivation-based methods. Due to its extremely wide range of application areas, fast metagenome sequencing simulation systems with high fidelity are in great demand to facilitate the development and comparison of metagenomics analysis tools. RESULTS: We present here a customizable metagenome simulation system: NeSSM (Next-generation Sequencing Simulator for Metagenomics. Combining complete genomes currently available, a community composition table, and sequencing parameters, it can simulate metagenome sequencing better than existing systems. Sequencing error models based on the explicit distribution of errors at each base and sequencing coverage bias are incorporated in the simulation. In order to improve the fidelity of simulation, tools are provided by NeSSM to estimate the sequencing error models, sequencing coverage bias and the community composition directly from existing metagenome sequencing data. Currently, NeSSM supports single-end and pair-end sequencing for both 454 and Illumina platforms. In addition, a GPU (graphics processing units version of NeSSM is also developed to accelerate the simulation. By comparing the simulated sequencing data from NeSSM with experimental metagenome sequencing data, we have demonstrated that NeSSM performs better in many aspects than existing popular metagenome simulators, such as MetaSim, GemSIM and Grinder. The GPU version of NeSSM is more than one-order of magnitude faster than MetaSim. CONCLUSIONS: NeSSM is a fast simulation system for high-throughput metagenome sequencing. It can be helpful to develop tools and evaluate strategies for metagenomics analysis and it's freely available for academic users at http://cbb.sjtu.edu.cn/~ccwei/pub/software/NeSSM.php.

  15. DAФNE Operation with Electron-Cloud-Clearing Electrodes

    CERN Document Server

    Alesini, D; Gallo, A; Guiducci, S; Milardi, C; Stella, A; Zobov, Mikhail; De Santis, S; Demma, Theo; Raimondi, P

    2013-01-01

    The effects of an electron cloud (e-cloud) on beam dynamics are one of the major factors limiting performances of high intensity positron, proton, and ion storage rings. In the electron-positron collider DAΦNE, namely, a horizontal beam instability due to the electron-cloud effect has been identified as one of the main limitations on the maximum stored positron beam current and as a source of beam quality deterioration. During the last machine shutdown in order to mitigate such instability, special electrodes have been inserted in all dipole and wiggler magnets of the positron ring. It has been the first installation all over the world of this type since long metallic electrodes have been installed in all arcs of the collider positron ring and are currently used during the machine operation in collision. This has allowed a number of unprecedented measurements (e-cloud instabilities growth rate, transverse beam size variation, tune shifts along the bunch train) where the e-cloud contribution is clearly eviden...

  16. Submarine pyroclastic deposits in Tertiary basins, NE Slovenia

    Directory of Open Access Journals (Sweden)

    Polona Kralj

    2013-12-01

    Full Text Available In Tertiary basins of NE Slovenia, Upper Oligocene volcanic activity occurred in a submarine environment that experienced contemporaneous clastic sedimentation. Pyroclastic deposits are essentially related to gas- and watersupported eruption-fed density currents. At Trobni Dol, the Lako Basin, an over 100 m thick deposit formed by a sigle sustained volcanic explosion that fed gas-supported pyroclastic flow. Diagnostic features are large matrixshard content, normal grading of pumice lapilli, collapsed pumice lapilli and the presence of charcoal. In the Smrekovec Volcanic Complex, several but only up to 5 m thick deposits related to eruption-fed gassupported pyroclastic flows occur. Deposits settled from water-supported eruption-fed density currents form fining- and thinning-upward sedimentary units which resemble the units of volcaniclastic turbidites. Pyroclastic deposits related to gas- and water-supported density currents occur in an up to 1000 m thick succession composed of coherent volcanics, autoclastic, pyroclastic, reworked volcaniclastic and mixed volcaniclastic-siliciclastic deposits that indicate a complex explosive and depositional history of the Smrekovec Volcanic Complex.

  17. Multi-usages of the Ilan geothermal field, NE Taiwan

    Science.gov (United States)

    Lee, C. S.; Tseng, P.; Wang, S.; Chang, C.

    2017-12-01

    The tectonics of Taiwan is very dynamic. The area produces more than 30,000 earthquakes/year; the mountains uplift 4-5 cm/year; the rainfall culminates 3,000 mm/year; there are some 4,000 hot spring operators. One of the two hot geothermal areas is located in NE Taiwan - the Ilan geothermal field. In order to develop the geothermal energy for the electricity need, the Ministry of Science and Technology have provided the fund to drill two 2,500 deep wells. The results are not so encourage for the need of an Enhanced Geothermal System. However, one of the wells has a bottom temperature of 160oC and the water up loading with 60 ton/hr. This can be combined with the near-by wells drilled by the private drilling company and the Cardinal Tien Junior College of Healthcare and Management to develop the multi-usages of the geothermal energy, such as 1 MW of electricity for the college and village, the long-term healthcare and hot spring medicare, aquaculture and agriculture need etc. The universities and private drilling company cooperate together to join the development. Hope this will provide a new model for the need of a self-sufficient community. The geothermal is a clean, renewable, and no pollution energy. Taiwan is in an initial stage of using this green energy.

  18. ASIC design in the KM3NeT detector

    International Nuclear Information System (INIS)

    Gajanana, D; Gromov, V; Timmer, P

    2013-01-01

    In the KM3NeT project [1], Cherenkov light from the muon interactions with transparent matter around the detector, is used to detect neutrinos. Photo multiplier tubes (PMT) used as photon sensor, are housed in a glass sphere (aka Optical Module) to detect single photons from the Cherenkov light. The PMT needs high operational voltage ( ∼ 1.5 kV) and is generated by a Cockroft-Walton (CW) multiplier circuit. The electronics required to control the PMT's and collect the signals is integrated in two ASIC's namely: 1) a front-end mixed signal ASIC (PROMiS) for the readout of the PMT and 2) an analog ASIC (CoCo) to generate pulses for charging the CW circuit and to control the feedback of the CW circuit. In this article, we discuss the two integrated circuits and test results of the complete setup. PROMiS amplifies the input charge, converts it to a pulse width and delivers the information via LVDS signals. These LVDS signals carry accurate information on the Time of arrival ( 2 C bus. This unique combination of the ASIC's results in a very cost and power efficient PMT base design.

  19. Nitrogen Uptake Rates during Spring in the NE Arabian Sea

    Directory of Open Access Journals (Sweden)

    Naveen Gandhi

    2010-01-01

    Full Text Available We present new data on N uptake rates and f-ratios in the north-eastern (NE Arabian Sea, where significant amounts of Trichodesmium were present in spring, 2006. The measured total nitrogen uptake rates ranged from 0.34 to 1.58 mmol N m−2d−1. N2 fixation associated with Trichodesmium varied from 0.002 to 0.54 mmol N m−2d−1 estimated from the abundance of Trichodesmium and specific N2 fixation rates of 1.5 pmol N trichome−1h−1. Inclusion of N2 fixation rates significantly changes f-ratios particularly in the coastal stations. Nitrogen isotopic data of surface suspended particles suggest that recently fixed nitrogen contributes as high as ~79% of the nitrogen in surface suspended particles. In addition, water column gained ~30 mmol N m−2 in the form of nitrate, likely due to nitrification of ammonium released by Trichodesmium. For better estimations, direct measurement of N2 fixation is recommended.

  20. Gels de poly(octylthiophène) : cristallisation

    Science.gov (United States)

    Pépin-Donat, B.; Sixou, B.; de Geyer, A.; Viallat, A.; Fah. Hin, L. Won

    1998-06-01

    The study of conjugated poly(octylthiophene) gels by x-ray diffraction and differential calorimetry shows that these networks are semi-crystalline. State of crystallinity, transport properties and large scale heterogeneity seem to be correlated. The determination of this correlation requires a detailed study of the mechanisms of crystallization of these polymer gels, the first results of which are reported here. They show that crystallinity ratios and crystallization rates can be determined by NMR and, as expected, strongly depend on the gels thermodynamic history. L'étude de gels conjugués de poly(octylthiophène) par diffraction des rayons X et par calorimétrie différentielle montre que ces réseaux sont semi-cristallins. Etat de cristallinité, propriétés de transport et état d'hétérogénéité à grande échelle semblent être corrélés. La détermination de cette corrélation nécessite une étude approfondie des mécanismes de cristallisation de ces gels polymères dont les premiers résultats sont reportés ici. Ils montrent que taux de cristallinité et vitesses de cristallisation peuvent être déterminés par RMN et, comme attendu, dépendent fortement de l'histoire thermodynamique des gels.

  1. Response matrices of NE213 scintillation detectors for neutrons

    Energy Technology Data Exchange (ETDEWEB)

    Guldbakke, S.; Klein, H. [Physikalisch-Technische Bundesanstalt, Braunschweig (Germany); Meister, A.; Scheler, U.; Unholzer, S. [Technical Univ., Dresden (Germany); Pulpan, J.; Tichy, M. [Inst. of Radiation Dosimetry, Prague (Czech Republic)

    1994-12-31

    Four NE213 detectors of different size have been calibrated at the accelerator facility of the PTB. The response functions were experimentally determined for 33 neutron energies between 1 MeV and 16 MeV and compared with Monte Carlo simulations using the NRESP7 code. The light output functions for recoil protons were found to be significantly different for all detectors even if they were of the same size. The neutron fluence determined on the basis of the response functions calculated with the corresponding light output functions agreed to better than {+-}2% with reference values if energy independent adjustment factors between 0.98 and 1.03 were applied. The response matrices required for the unfolding of neutron induced pulse height spectra were therefore calculated with the NRESP7 code taking into account the adjustment factors. Similarly, the response matrices for photons were calculated with the EGS4 code, but without any adjustment. Finally, the DIFBAS code was applied for the unfolding of pure neutron- and photon-induced pulse height spectra. The resulting spectral fluences are in reasonable agreement with the results obtained by time-of-flight measurements and by spectrometry with a Ge detector.

  2. Response matrices of NE213 scintillation detectors for neutrons

    International Nuclear Information System (INIS)

    Guldbakke, S.; Klein, H.; Meister, A.; Scheler, U.; Unholzer, S.; Pulpan, J.; Tichy, M.

    1994-01-01

    Four NE213 detectors of different size have been calibrated at the accelerator facility of the PTB. The response functions were experimentally determined for 33 neutron energies between 1 MeV and 16 MeV and compared with Monte Carlo simulations using the NRESP7 code. The light output functions for recoil protons were found to be significantly different for all detectors even if they were of the same size. The neutron fluence determined on the basis of the response functions calculated with the corresponding light output functions agreed to better than ±2% with reference values if energy independent adjustment factors between 0.98 and 1.03 were applied. The response matrices required for the unfolding of neutron induced pulse height spectra were therefore calculated with the NRESP7 code taking into account the adjustment factors. Similarly, the response matrices for photons were calculated with the EGS4 code, but without any adjustment. Finally, the DIFBAS code was applied for the unfolding of pure neutron- and photon-induced pulse height spectra. The resulting spectral fluences are in reasonable agreement with the results obtained by time-of-flight measurements and by spectrometry with a Ge detector

  3. Vloga teorije gestalt v snovanju in oblikovanju celotne grafične podobe

    OpenAIRE

    Mesarič Bohar, Nina

    2017-01-01

    Poznavanje gestalt principov je za oblikovalca poznavanje dragocenih oblikovalskih orodij. Občinstvo bo vedno odreagiralo na očitne gestalt vzorce. Z ujemanjem ciljnega občinstva z izbranimi gestalt deli lahko skrajšamo pot do efektivnega komuniciranja. Gestalt principi opisujejo različne načine, kako ljudje individualne elemente grupirajo skupaj. Pri oblikovanju celotne grafične podobe in spletne strani je pomembno, da upoštevamo celotne elemente in ne le njene posamezne elemente, ker še...

  4. Čelične okvirne konstrukcije nasuprot samonosivim regalnim skladištima

    OpenAIRE

    Vujanac, Rodoljub; Živković, Miroslav; Slavković, Radovan; Vulović, Snežana

    2017-01-01

    Čelične okvirne konstrukcije hala sa stupovima i krovnim gredama ili rešetkama su najzastupljeniji stil građenja skladišnih hala za proizvodne i distributivne potrebe. Ovisno o namjeni, klasične skladišne hale su po potrebi opremljene različitim tipovima regalne konstrukcije. Konstrukcija samonosećeg paletnog regalnog skladišta, kao poseban vid montažnog objekta, uglavnom se sastoji od elemenata regalnog sustava koji nosi krov i zidove, a istovremeno služi i za skladištenje materijala. Svi el...

  5. Neutrinos from LHC and the Mediterranean very large neutrino telescope (KM3NeT)

    Science.gov (United States)

    Shanidze, Rezo

    2006-11-01

    High-energy neutrinos will be copiously produced from the proton beams of the Large Hadron Collider (LHC) at CERN. We consider neutrino fluxes from the LHC and estimate possible event rates for a future very large volume neutrino telescope in the Mediterranean Sea (KM3NeT). The rates were obtained for the case when the LHC neutrinos are directed to the neutrino telescope, although in the current configuration neutrino fluxes are not pointing to the possible KM3NeT sites. Availability of the artificial and controllable source of neutrinos could significantly enhance the physics potential of the KM3NeT project.

  6. Detector optimisation studies for the KM3NeT conceptual design report

    Energy Technology Data Exchange (ETDEWEB)

    Shanidze, Rezo [Erlangen Centre for Astroparticle Physics (ECAP), Erlangen University (Germany)

    2008-07-01

    The KM3NeT design study is an EU-funded project for an European deep-sea research infrastructure, which will host a high energy neutrino telescope with a volume of at least one cubic kilometre in the Mediterranean Sea. Recently, the KM3NeT consortium prepared the conceptual design report (CDR), in which the possible options for the KM3NeT detector are described. The detector optimisation studies, performed in Erlangen using MC simulations, are presented in the talk.

  7. Neutrinos from LHC and the Mediterranean very large neutrino telescope (KM3NeT)

    Energy Technology Data Exchange (ETDEWEB)

    Shanidze, Rezo [Physics Institute, University of Erlangen, Erwin-Rommel Str.1, D-91058 Erlangen (Germany)]. E-mail: shanidze@physik.uni-erlangen.de

    2006-11-15

    High-energy neutrinos will be copiously produced from the proton beams of the Large Hadron Collider (LHC) at CERN. We consider neutrino fluxes from the LHC and estimate possible event rates for a future very large volume neutrino telescope in the Mediterranean Sea (KM3NeT). The rates were obtained for the case when the LHC neutrinos are directed to the neutrino telescope, although in the current configuration neutrino fluxes are not pointing to the possible KM3NeT sites. Availability of the artificial and controllable source of neutrinos could significantly enhance the physics potential of the KM3NeT project.

  8. Statična analiza večetažne zidane stavbe v Hrpeljah

    OpenAIRE

    Mugerli, Matija

    2017-01-01

    V diplomski nalogi sem analiziral in dimenzioniral karakteristične elemente nosilne konstrukcije večetažne zidane stanovanjske stavbe v Hrpeljah. Vsebinsko je naloga sestavljena iz dveh delov in sicer iz analize nosilnosti medetažne armiranobetonske (AB) plošče ter iz analize nosilnosti zidanih sten v pritličju stavbe. Pri analizi sem se poslužil metod mejnih stanj in sicer mejnih stanj nosilnosti (MSN) in mejnih stanj uporabnosti (MSU). Analizo AB plošče sem izvedel s pomočjo idealizira...

  9. POVEČANJE IZKORISTKA PRIDOBIVANJA SONČNE ENERGIJE Z AKTIVNIM SLEDENJEM

    OpenAIRE

    Bračič, Simon

    2009-01-01

    V diplomskem delu bomo predstavili nekaj o virih energije (obnovljivi in neobnovljivi viri) in se podrobneje posvetili sončni energiji. Raziskali bomo, kakšne so trenutne možnosti izkoriščanja sončne energije in ali je trenutno nakup sistema za izkoriščanje te energije rentabilen. Podrobneje bomo raziskali, kako lahko povečamo izkoristek sončne energije s sledenjem Soncu in tudi sami izdelali napravo za povečanje izkoristka sončnih celic ali kolektorjev.

  10. Measurements of low-energy e+--Ne and e+-Ar total scattering cross sections

    International Nuclear Information System (INIS)

    Charlton, M.; Laricchia, G.; Griffith, T.C.; Wright, G.L.; Heyland, G.R.

    1984-01-01

    Measurements of positron and electron-Ne total scattering cross sections are reported in the energy range 2-50 eV. e + -Ar total cross sections are presented in the energy range 2-20 eV. In both the e + -Ne and e + -Ar cases marked discrepancies are found with other recent measurements and these are discussed along with possible systematic errors which may affect the data. The e - -Ne measurements are found to be in excellent agreement with recent experiments. (author)

  11. The Jurassic of Denmark and Greenland: The Jurassic of Skåne, southern Sweden

    OpenAIRE

    Sivhed, Ulf; Ahlberg, Anders; Erlström, Mikael

    2003-01-01

    In Sweden, Jurassic strata are restricted to Skåne and adjacent offshore areas. Jurassic sedimentary rocks predominantly comprise sandy to muddy siliciclastics, with subordinate coal beds andfew carbonate-rich beds. During Mesozoic times, block-faulting took place in the Sorgenfrei–Tornquist Zone, a tectonic zone which transects Skåne in a NW–SE direction. The Jurassic depositionalenvironments in Skåne were thus strongly influenced by uplift and downfaulting, and to some extent by volcanism. ...

  12. KM3NeT/ARCA sensitivity and discovery potential for neutrino point-like sources

    Directory of Open Access Journals (Sweden)

    Trovato A.

    2016-01-01

    Full Text Available KM3NeT is a large research infrastructure with a network of deep-sea neutrino telescopes in the abyss of the Mediterranean Sea. Of these, the KM3NeT/ARCA detector, installed in the KM3NeT-It node of the network, is optimised for studying high-energy neutrinos of cosmic origin. Sensitivities to galactic sources such as the supernova remnant RXJ1713.7-3946 and the pulsar wind nebula Vela X are presented as well as sensitivities to a generic point source with an E−2 spectrum which represents an approximation for the spectrum of extragalactic candidate neutrino sources.

  13. Excitation functions for some Ne induced reactions with Holmium: incomplete fusion vs complete fusion

    International Nuclear Information System (INIS)

    Agarwal, Avinash; Kumar, Munish; Sharma, Anjali; Rizvi, I.A.; Ahamad, Tauseef; Ghugre, S.S.; Sinha, A.K.; Chaubey, A.K.

    2010-01-01

    Reactions induced by 20 Ne are expected to be considerably more complex than those of 12 C, and 16 O. As a part of the ongoing program to understand CF and ICF reaction mechanisms, it is of great interest to see whether the same experimental technique yield similarly valuable information for 20 Ne induced reactions. In this present work an attempt has been made to measure the excitation functions for fifteen evaporation residues (ERs) identified in the interaction of 20 Ne + 165 Ho system in the energy range 4 -7 MeV/A

  14. Gamma-decay study of sup 2 sup 1 Na and sup 2 sup 1 Ne, octupole bands in sup 2 sup 1 Ne

    CERN Document Server

    Thummerer, S; Kokalova, T; Bohlen, H G; Gebauer, B; Tumino, A; Massey, T N; Angelis, G D; Axiotis, M; Gadea, A; Kröll, T; Marginean, N; Napoli, D R; Poli, M; Ur, C; Bazzacco, D; Lenzi, S M; Alvarez, C R; Lunardi, S; Menegazzo, R; Bizzeti, P G; Bizzeti-Sona, A M

    2003-01-01

    The reactions sup 1 sup 6 O( sup 7 Li, 2n) and sup 1 sup 6 O( sup 7 Li, np) populating sup 2 sup 1 Na and sup 2 sup 1 Ne have been studied at E sub L = 27 MeV using the GASP gamma-detector array. The level scheme for sup 2 sup 1 Na and sup 2 sup 1 Ne has been extended to higher excitation energy. Spins and parities of the observed levels are assigned tentatively supporting the identification of opposite parity structures connected by strong dipole transitions. The structure of these bands is interpreted as based on a reflection asymmetric cluster structure.

  15. Observation of an impact-parameter window in low-velocity ionizing collisions of Ne+ on Ne proceeding through quasimolecular states

    International Nuclear Information System (INIS)

    Abdallah, M.A.; Cocke, C.L.; Stoeckli, M.; Wolff, W.; Wolf, H.E.

    1998-01-01

    Target ionization in collisions of singly charged Ne + ions with Ne has been investigated at projectile velocities from 0.25 to 0.55 a.u. using electron and recoil momentum imaging techniques. The momentum distributions of the ejected electrons were found to carry a distinct signature strongly suggesting that ionization is taking place by successive promotions through molecular orbitals. The observed recoil transverse momentum distributions are donut-shaped, indicating that single ionization is confined to a well-defined impact-parameter window. copyright 1998 The American Physical Society

  16. Alimentation du nouveau-ne et du nourrisson dans la region ...

    African Journals Online (AJOL)

    Alimentation du nouveau-ne et du nourrisson dans la region centrale du togo : pratiques familiales et communautaires avant la mise en oeuvre de la strategie « prise en charge integree des maladies de l'enfant »

  17. Lääne abi vajava Moldova toetamine on Eesti huvides / Marianne Mikko

    Index Scriptorium Estoniae

    Mikko, Marianne, 1961-

    2004-01-01

    Euroopa Parlamendi Moldova delegatsiooni juht annab ülevaate olukorrast riigis. Autori sõnul nähakse Moldovas Transnistria probleemi lahendamise ainsat võimalust Lääne toetuses ning surves Ukrainale

  18. Momentum dependence of hadrons produced in ν(antiν)Ne interactions

    International Nuclear Information System (INIS)

    Lubatti, H.J.; Burnett, T.H.; Csorna, S.E.; Holmgren, D.; Kollman, G.; Moriyasu, K.; Rudnicka, H.; Swider, G.M.; Yuldashev, B.S.; Ballagh, H.C.; Bingham, H.H.; Fretter, W.B.; Gee, D.; Lynch, G.; Marriner, J.P.; Orthel, J.; Stevenson, M.L.; Yost, G.P.

    The inclusive distributions of hadrons produced in ν(anti ν)Ne interactions are studied as a function of the momentum fraction z. Discrepancies between the data and existing parametrizations of the quark-parton model are found. (author)

  19. The neXtProt knowledgebase on human proteins: current status.

    Science.gov (United States)

    Gaudet, Pascale; Michel, Pierre-André; Zahn-Zabal, Monique; Cusin, Isabelle; Duek, Paula D; Evalet, Olivier; Gateau, Alain; Gleizes, Anne; Pereira, Mario; Teixeira, Daniel; Zhang, Ying; Lane, Lydie; Bairoch, Amos

    2015-01-01

    neXtProt (http://www.nextprot.org) is a human protein-centric knowledgebase developed at the SIB Swiss Institute of Bioinformatics. Focused solely on human proteins, neXtProt aims to provide a state of the art resource for the representation of human biology by capturing a wide range of data, precise annotations, fully traceable data provenance and a web interface which enables researchers to find and view information in a comprehensive manner. Since the introductory neXtProt publication, significant advances have been made on three main aspects: the representation of proteomics data, an extended representation of human variants and the development of an advanced search capability built around semantic technologies. These changes are presented in the current neXtProt update. © The Author(s) 2014. Published by Oxford University Press on behalf of Nucleic Acids Research.

  20. Arisaema gracilentum, a new species of Arisaema (Araceae) from NE India

    NARCIS (Netherlands)

    Bruggeman, P.

    2016-01-01

    Arisaema gracilentum, a new species of Araceae, belonging to section Arisaema from the Lower Dibang Valley, Arunachal Pradesh State in NE India is described, illustrated and compared with related taxa.

  1. Spectral line intensities of NeVII for non-equilibrium ionization plasma including dielectronic recombination processes

    Energy Technology Data Exchange (ETDEWEB)

    Murakami, Izumi; Kato, Takako [National Inst. for Fusion Science, Toki, Gifu (Japan); Safronova, U.

    1999-01-01

    We have calculated the dielectronic recombination rate coefficients from Li-like Ne (Ne{sup 7+}) ions to Be-like Ne (Ne{sup 6+}) ions for selected excited states of Ne{sup 6+} ions. A collisional-radiative model (CRM) for Ne{sup 6+} ions is constructed to calculate the population density of each excited state in non-equilibrium ionization plasmas, including recombining processes. NeVII spectral line intensities and the radiative power loss are calculated with the CRM. A density effect caused by collisional excitation from the metastable state 2s2p {sup 3}P is found at an electron density of 10{sup 5} - 10{sup 17} cm{sup -3}. The collisional excitations between excited states become important at high electron temperature T{sub e} > or approx. 100 eV. (author)

  2. Mitigation of gamma-radiation induced abasic sites in genomic DNA by dietary nicotinamide supplementation: Metabolic up-regulation of NAD{sup +} biosynthesis

    Energy Technology Data Exchange (ETDEWEB)

    Batra, Vipen, E-mail: batravipen@gmail.com; Kislay, Binita

    2013-09-15

    Highlights: • Dietary nicotinamide increases enzyme dependent NAD{sup +} synthesis after irradiation. • Enhanced NAD{sup +} levels mitigate gamma (γ)-radiation induced abasic sites in DNA. • Dietary nicotinamide induces and prolongs expression of excision repair enzymes. • Nicotinamide reduces radiation-generated biomarker (8-oxo-dG) of DNA base damage. • Dietary nicotinamide reduces radiation inflicted DNA damage and delays apoptosis. - Abstract: The search for non-toxic radio-protective drugs has yielded many potential agents but most of these compounds have certain amount of toxicity. The objective of the present study was to investigate dietary nicotinamide enrichment dependent adaptive response to potential cytotoxic effect of {sup 60}Co γ-radiation. To elucidate the possible underlying mechanism(s), male Swiss mice were maintained on control diet (CD) and nicotinamide supplemented diet (NSD). After 6 weeks of CD and NSD dietary regimen, we exposed the animals to γ-radiation (2, 4 and 6 Gy) and investigated the profile of downstream metabolites and activities of enzymes involved in NAD{sup +} biosynthesis. Increased activities of nicotinamide phosphoribosyltransferase (NAMPT) and nicotinamide mononucleotide adenylyltransferase (NMNAT) were observed up to 48 h post-irradiation in NSD fed irradiated mice. Concomitant with increase in liver NAMPT and NMNAT activities, NAD{sup +} levels were replenished in NSD fed and irradiated animals. However, NAMPT and NMNAT-mediated NAD{sup +} biosynthesis and ATP levels were severely compromised in liver of CD fed irradiated mice. Another major finding of these studies revealed that under γ-radiation stress, dietary nicotinamide supplementation might induce higher and long-lasting poly(ADP)-ribose polymerase 1 (PARP1) and poly(ADP-ribose) glycohydrolase (PARG) activities in NSD fed animals compared to CD fed animals. To investigate liver DNA damage, number of apurinic/apyrimidinic sites (AP sites) and level of

  3. Reservoir-induced seismicity at Castanhao reservoir, NE Brazil

    Science.gov (United States)

    Nunes, B.; do Nascimento, A.; Ferreira, J.; Bezerra, F.

    2012-04-01

    Our case study - the Castanhão reservoir - is located in NE Brazil on crystalline rock at the Borborema Province. The Borborema Province is a major Proterozoic-Archean terrain formed as a consequence of convergence and collision of the São Luis-West Africa craton and the São Francisco-Congo-Kasai cratons. This reservoir is a 60 m high earth-filled dam, which can store up to 4.5 billion m3 of water. The construction begun in 1990 and finished in October 2003.The first identified reservoir-induced events occurred in 2003, when the water level was still low. The water reached the spillway for the first time in January 2004 and, after that, an increase in seismicity occured. The present study shows the results of a campaign done in the period from November 19th, 2009 to December 31th, 2010 at the Castanhão reservoir. We deployed six three-component digital seismographic station network around one of the areas of the reservoir. We analyzed a total of 77 events which were recorded in at least four stations. To determine hypocenters and time origin, we used HYPO71 program (Lee & Lahr, 1975) assuming a half-space model with following parameters: VP= 5.95 km/s and VP/VS=1.73. We also performed a relocation of these events using HYPODD (Waldhauser & Ellsworth, 2000) programme. The input data used we used were catalogue data, with all absolute times. The results from the spatio-temporal suggest that different clusters at different areas and depths are triggered at different times due to a mixture of: i - pore pressure increase due to diffusion and ii - increase of pore pressure due to the reservoir load.

  4. Phylogeographic analysis of paternal lineages in NE Portuguese Jewish communities.

    Science.gov (United States)

    Nogueiro, Inês; Manco, Licínio; Gomes, Verónica; Amorim, António; Gusmão, Leonor

    2010-03-01

    The establishment of Jewish communities in the territory of contemporary Portugal is archaeologically documented since the 3rd century CE, but their settlement in Trás-os-Montes (NE Portugal) has not been proved before the 12th century. The Decree of Expulsion followed by the establishment of the Inquisition, both around the beginning of the 16th century, accounted for a significant exodus, as well as the establishment of crypto-Jewish communities. Previous Y chromosome studies have shown that different Jewish communities share a common origin in the Near East, although they can be quite heterogeneous as a consequence of genetic drift and different levels of admixture with their respective host populations. To characterize the genetic composition of the Portuguese Jewish communities from Trás-os-Montes, we have examined 57 unrelated Jewish males, with a high-resolution Y-chromosome typing strategy, comprising 16 STRs and 23 SNPs. A high lineage diversity was found, at both haplotype and haplogroup levels (98.74 and 82.83%, respectively), demonstrating the absence of either strong drift or founder effects. A deeper and more detailed investigation is required to clarify how these communities avoided the expected inbreeding caused by over four centuries of religious repression. Concerning haplogroup lineages, we detected some admixture with the Western European non-Jewish populations (R1b1b2-M269, approximately 28%), along with a strong ancestral component reflecting their origin in the Middle East [J1(xJ1a-M267), approximately 12%; J2-M172, approximately 25%; T-M70, approximately 16%] and in consequence Trás-os-Montes Jews were found to be more closely related with other Jewish groups, rather than with the Portuguese non-Jewish population.

  5. Diffuse fluxes of cosmic neutrinos in KM3NeT

    Science.gov (United States)

    Seitz, T.; Shanidze, R.; KM3NeT Consortium1

    2011-01-01

    The search for diffuse fluxes of cosmic neutrinos is among the main objectives of high-energy neutrino astronomy. The future Mediterranean deep-sea neutrino telescope KM3NeT, due to its multi-km3 instrumented volume will have unprecedented sensitivity for the observation of these fluxes. Detection of diffuse neutrino fluxes of different origins (extragalactic, from inner Galaxy and cosmogenic) in the KM3NeT neutrino telescope is discussed in this paper.

  6. Diffuse fluxes of cosmic neutrinos in KM3NeT

    Energy Technology Data Exchange (ETDEWEB)

    Seitz, T., E-mail: thomas.seitz@physik.uni-erlangen.d [Erlangen Centre for Astroparticle Physics, University of Erlangen-Nuernberg, Erwin-Rommel-Str. 1, 91058 Erlangen (Germany); Shanidze, R., E-mail: shanidze@physik.uni-erlangen.d [Erlangen Centre for Astroparticle Physics, University of Erlangen-Nuernberg, Erwin-Rommel-Str. 1, 91058 Erlangen (Germany)

    2011-01-21

    The search for diffuse fluxes of cosmic neutrinos is among the main objectives of high-energy neutrino astronomy. The future Mediterranean deep-sea neutrino telescope KM3NeT, due to its multi-km{sup 3} instrumented volume will have unprecedented sensitivity for the observation of these fluxes. Detection of diffuse neutrino fluxes of different origins (extragalactic, from inner Galaxy and cosmogenic) in the KM3NeT neutrino telescope is discussed in this paper.

  7. Agnostic stacking of intergalactic doublet absorption: measuring the Ne VIII population

    Science.gov (United States)

    Frank, Stephan; Pieri, Matthew M.; Mathur, Smita; Danforth, Charles W.; Shull, J. Michael

    2018-05-01

    We present a blind search for doublet intergalactic metal absorption with a method dubbed `agnostic stacking'. Using a forward-modelling framework, we combine this with direct detections in the literature to measure the overall metal population. We apply this novel approach to the search for Ne VIII absorption in a set of 26 high-quality COS spectra. We probe to an unprecedented low limit of log N>12.3 at 0.47≤z ≤1.34 over a path-length Δz = 7.36. This method selects apparent absorption without requiring knowledge of its source. Stacking this mixed population dilutes doublet features in composite spectra in a deterministic manner, allowing us to measure the proportion corresponding to Ne VIII absorption. We stack potential Ne VIII absorption in two regimes: absorption too weak to be significant in direct line studies (12.3 13.7). We do not detect Ne VIII absorption in either regime. Combining our measurements with direct detections, we find that the Ne VIII population is reproduced with a power-law column density distribution function with slope β = -1.86 ^{+0.18 }_{ -0.26} and normalization log f_{13.7} = -13.99 ^{+0.20 }_{ -0.23}, leading to an incidence rate of strong Ne VIII absorbers dn/dz =1.38 ^{+0.97 }_{ -0.82}. We infer a cosmic mass density for Ne VIII gas with 12.3 < log N < 15.0 of Ω _{{{Ne {VIII}}}} = 2.2 ^{+1.6 }_{ _-1.2} × 10^{-8}, a value significantly lower that than predicted by recent simulations. We translate this density into an estimate of the baryon density Ωb ≈ 1.8 × 10-3, constituting 4 per cent of the total baryonic mass.

  8. Agnostic Stacking of Intergalactic Doublet Absorption: Measuring the Ne VIII Population

    Science.gov (United States)

    Frank, Stephan; Pieri, Matthew M.; Mathur, Smita; Danforth, Charles W.; Michael Shull, J.

    2018-02-01

    We present a blind search for doublet intergalactic metal absorption with a method dubbed `agnostic stacking'. Using a forward-modelling framework we combine this with direct detections in the literature to measure the overall metal population. We apply this novel approach to the search for Ne VIII absorption in a set of 26 high-quality COS spectra. We probe to an unprecedented low limit of log N>12.3 at 0.47≤z ≤1.34 over a pathlength Δz = 7.36. This method selects apparent absorption without requiring knowledge of its source. Stacking this mixed population dilutes doublet features in composite spectra in a deterministic manner, allowing us to measure the proportion corresponding to Ne VIII absorption. We stack potential Ne VIII absorption in two regimes: absorption too weak to be significant in direct line studies (12.3 13.7). We do not detect Ne VIII absorption in either regime. Combining our measurements with direct detections, we find that the Ne VIII population is reproduced with a power law column density distribution function with slope β = -1.86+0.18-0.26 and normalisation log f_{13.7} = -13.99+0.20-0.23, leading to an incidence rate of strong Ne VIII absorbers dn/dz =1.38+0.97-0.82. We infer a cosmic mass density for Ne VIII gas with 12.3 < log N < 15.0 of Ω _{Ne VIII} = 2.2+1.6-1.2 × 10^{-8}, a value significantly lower that than predicted by recent simulations. We translate this density into an estimate of the baryon density Ωb ≈ 1.8 × 10-3, constituting 4% of the total baryonic mass.

  9. [Ne III]/[O II] as an ionization parameter diagnostic in star-forming galaxies

    International Nuclear Information System (INIS)

    Levesque, Emily M.; Richardson, Mark L. A.

    2014-01-01

    We present our parameterizations of the log([Ne III]λ3869/[O II]λ3727) (Ne3O2) and log([O III]λ5007/[O II]λ3727) (O3O2) ratios as diagnostics of ionization parameter in star-forming galaxies. Our calibrations are based on the Starburst99/Mappings III photoionization models, which extend up to the extremely high values of ionization parameter found in high-redshift galaxies. While similar calibrations have been presented previously for O3O2, this is the first such calibration of Ne3O2. We illustrate the tight correlation between these two ratios for star-forming galaxies and discuss the underlying physics that dictates their very similar evolution. Based on this work, we propose the Ne3O2 ratio as a new and useful diagnostic of ionization parameter for star-forming galaxies. Given the Ne3O2 ratio's relative insensitivity to reddening, this ratio is particularly valuable for use with galaxies that have uncertain amounts of extinction. The short wavelengths of the Ne3O2 ratio can also be applied out to very high redshifts, extending studies of galaxies' ionization parameters out to z ∼ 1.6 with optical spectroscopy and z ∼ 5.2 with ground-based near-infrared spectra.

  10. Could LC-NE-Dependent Adjustment of Neural Gain Drive Functional Brain Network Reorganization?

    Directory of Open Access Journals (Sweden)

    Carole Guedj

    2017-01-01

    Full Text Available The locus coeruleus-norepinephrine (LC-NE system is thought to act at synaptic, cellular, microcircuit, and network levels to facilitate cognitive functions through at least two different processes, not mutually exclusive. Accordingly, as a reset signal, the LC-NE system could trigger brain network reorganizations in response to salient information in the environment and/or adjust the neural gain within its target regions to optimize behavioral responses. Here, we provide evidence of the co-occurrence of these two mechanisms at the whole-brain level, in resting-state conditions following a pharmacological stimulation of the LC-NE system. We propose that these two mechanisms are interdependent such that the LC-NE-dependent adjustment of the neural gain inferred from the clustering coefficient could drive functional brain network reorganizations through coherence in the gamma rhythm. Via the temporal dynamic of gamma-range band-limited power, the release of NE could adjust the neural gain, promoting interactions only within the neuronal populations whose amplitude envelopes are correlated, thus making it possible to reorganize neuronal ensembles, functional networks, and ultimately, behavioral responses. Thus, our proposal offers a unified framework integrating the putative influence of the LC-NE system on both local- and long-range adjustments of brain dynamics underlying behavioral flexibility.

  11. The Jurassic of Denmark and Greenland: The Jurassic of Skåne, southern Sweden

    Directory of Open Access Journals (Sweden)

    Sivhed, Ulf

    2003-10-01

    Full Text Available In Sweden, Jurassic strata are restricted to Skåne and adjacent offshore areas. Jurassic sedimentary rocks predominantly comprise sandy to muddy siliciclastics, with subordinate coal beds andfew carbonate-rich beds. During Mesozoic times, block-faulting took place in the Sorgenfrei–Tornquist Zone, a tectonic zone which transects Skåne in a NW–SE direction. The Jurassic depositionalenvironments in Skåne were thus strongly influenced by uplift and downfaulting, and to some extent by volcanism. Consequently, the sedimentary record reveals evidence of numerous transgressions, regressions and breaks in sedimentation. Relative sea-level changes played a significant role in controlling the facies distribution, as deposition mainly took place in coastal plain to shallow shelf environments.The alluvial deposits in Skåne include floodplain palaeosols, autochthonous coals, overbank sandstones, and stream channel pebbly sandstones. Restricted marine strata comprise intertidalheteroliths with mixed freshwater and marine trace fossil assemblages, and intertidal delta distributary channel sandstones. Shallow marine sediments encompass subtidal and shoreface sandstoneswith herringbone structures, and bioturbated mudstones with tempestite sandstones. Offshore deposits typically comprise extensively bioturbated muddy sandstones.Floral remains, palaeopedology, clay mineralogy and arenite maturity indicate a warm and humid climate in Skåne throughout the Jurassic, possibly with slightly increasing aridity towards the end of the period. Most Jurassic strata in Skåne have been subjected to mild burial diagenesis, and the petroleum generative window has rarely been reached.

  12. Matériaux pour la production d'hydrogène

    Science.gov (United States)

    Saint-Just, J.

    2002-04-01

    L'émergence des piles à combustible et l'apparition de l'hydrogène comme carburant dans des véhicules grand public comme les autobus, suscitent un élan fort pour la mise au point de technologies nouvelles pour la production d'hydrogène décentralisée et à petite échelle. Du fait de sa richesse en hydrogène. des réserves disponibles et de son infrastructure de distribution, le gaz naturel est considéré comme la ressource majeure pouvant fournir les quantités d'hydrogène nécessaires pour un passage progressif vers une économie de l'hydrogène dans les décennies immédiates en attendant que progressivement les énergies renouvelables prennent le relais. Des solutions innovantes, incluant de nouveaux matériaux, sont nécessaires pour la mise au point des petits générateurs d'hydrogène efficaces et bon marché alimenté au gaz naturel qui sont attendus par les fabricants (le piles à combustible, les acteurs de l'énergie et les fabricants de véhicules.

  13. Stabilization of Au Monatomic-High Islands on the (2 ×2 )-Nad Reconstructed Surface of Wurtzite AlN(0001)

    Science.gov (United States)

    Eydoux, Benoit; Baris, Bulent; Khoussa, Hassan; Guillermet, Olivier; Gauthier, Sébastien; Bouju, Xavier; Martrou, David

    2017-10-01

    Noncontact atomic force microscopy images show that gold grows on the (2 ×2 )-Nad reconstructed polar (0001) surface of AlN insulating films, in the form of large monatomic islands. High-resolution images and in situ reflection high-energy electron diffraction spectra reveal two moiré patterns from which an atomic model can be built. Density functional theory calculations confirm this model and give insight into the mechanisms that lead to the stabilization of the monolayer. Gold adsorption is accompanied, first, by a global vertical charge transfer from the AlN substrate that fulfills the electrostatic stability criterion for a polar material, and second, by lateral charge transfers that are driven by the local chemical properties of the (2 ×2 )-Nad reconstruction. These results present alternative strategies to grow metal electrodes onto nitride compounds with a better controlled interface, a crucial issue for applications.

  14. Expression, purification, crystallization and preliminary X-ray analysis of an NAD-dependent glyceraldehyde-3-phosphate dehydrogenase from Helicobacter pylori

    Energy Technology Data Exchange (ETDEWEB)

    Elliott, Paul R.; Mohammad, Shabaz; Melrose, Helen J.; Moody, Peter C. E., E-mail: pcem1@leicester.ac.uk [Henry Wellcome Laboratories for Structural Biology, University of Leicester, Leicester LE1 9HN (United Kingdom)

    2008-08-01

    Glyceraldehyde-3-phosphate dehydrogenase B from H. pylori has been cloned, expressed, purified and crystallized in the presence of NAD. Crystals of GAPDHB diffracted to 2.8 Å resolution and belonged to space group P6{sub 5}22, with unit-cell parameters a = b = 166.1, c = 253.1 Å. Helicobacter pylori is a dangerous human pathogen that resides in the upper gastrointestinal tract. Little is known about its metabolism and with the onset of antibiotic resistance new treatments are required. In this study, the expression, purification, crystallization and preliminary X-ray diffraction of an NAD-dependent glyceraldehyde-3-phosphate dehydrogenase from H. pylori are reported.

  15. Biochemical characterisation of LigN, an NAD+-dependent DNA ligase from the halophilic euryarchaeon Haloferax volcanii that displays maximal in vitro activity at high salt concentrations

    DEFF Research Database (Denmark)

    Poidevin, L.; MacNeill, S. A.

    2006-01-01

    Background DNA ligases are required for DNA strand joining in all forms of cellular life. NAD+-dependent DNA ligases are found primarily in eubacteria but also in some eukaryotic viruses, bacteriophage and archaea. Among the archaeal NAD+-dependent DNA ligases is the LigN enzyme of the halophilic...... assays using ¿ DNA restriction fragments with 12 bp cos cohesive ends were used to show that LigN activity was dependent on addition of divalent cations and salt. No activity was detected in the absence of KCl, whereas maximum activity could be detected at 3.2 M KCl, close to the intracellular KCl...... concentration of Hfx.volcanii cells. Conclusion LigN is unique amongst characterised DNA ligase enzymes in displaying maximal DNA strand joining activity at high (> 3 M) salt levels. As such the LigN enzyme has potential both as a novel tool for biotechnology and as a model enzyme for studying the adaptation...

  16. Immunohistochemical Analysis of Neuroendocrine (NE) Differentiation in Testicular Germ Cell Tumors (GCTs): Use of Confocal Laser Scanning Microscopy (CLSM) to Demonstrate Direct NE Differentiation from GCTs

    International Nuclear Information System (INIS)

    Kumaki, Nobue; Umemura, Shinobu; Kajiwara, Hiroshi; Itoh, Johbu; Itoh, Yoshiko; Osamura, R.Yoshiyuki

    2007-01-01

    Neuroendocrine (NE) differentiation is infrequent in testicular tumors and its histogenesis is not well understood. The present study is aimed at elucidating the pathway of neuroendocrine differentiation in germ cell tumors (GCTs) of the testis. In the analysis of 46 germ cell tumor components from 23 testicular tumors, we focused on GCTs with neuroendocrine differentiation, 7 teratoma, 1 embryonal carcinoma and 1 neuroendocrine carcinoma by immunohistochemical study and confocal laser scanning microscopy (CLSM) analysis. NE marker positive cells were noted in the tumor with collision of teratoma and embryonal carcinoma (E&T tumor), in the immature columnar cells of transitional form of embryonal carcinoma to teratoma (E-T cells) and neuroendocrine carcinoma cells, in addition to the well known mature intestinal mucosa in teratoma. Double staining for a NE marker (CGA) and a germ cell marker (PLAP) demonstrated the localization of both proteins in the same E-T cells confirmed by CLSM. Another finding, indicating the intimate relation between embryonal carcinoma and neuroendcrine differentiation, is that neuroendocrine carcinoma expressed a marker of embryonal carcinoma, CD30. The present results indicated that the NE cells might be differentiated from embryonal carcinoma, a view that has not been proposed before, but that is made in the present study using CLSM

  17. Deep sea tests of a prototype of the KM3NeT digital optical module : KM3NeT Collaboration

    NARCIS (Netherlands)

    Adrián-Martínez, S.; Ageron, M.; Aharonian, F.; Aiello, S.; Albert, A.; Ameli, F.; Anassontzis, E. G.; Anghinolfi, M.; Anton, G.; Anvar, S.; Ardid, M.; de Asmundis, R.; Balasi, K.; Band, H.; Barbarino, G.; Barbarito, E.; Barbato, F.; Baret, B.; Baron, S.; Belias, A.; Berbee, E.; van den Berg, A. M.; Berkien, A.; Bertin, V.; Beurthey, S.; van Beveren, V.; Beverini, N.; Biagi, S.; Bianucci, S.; Billault, M.; Birbas, A.; Boer Rookhuizen, H.; Bormuth, R.; Bouché, V.; Bouhadef, B.; Bourlis, G.; Bouwhuis, M.; Bozza, C.; Bruijn, R.; Brunner, J.; Cacopardo, G.; Caillat, L.; Calamai, M.; Calvo, D.; Capone, A.; Caramete, L.; Caruso, F.; Cecchini, S.; Ceres, A.; Cereseto, R.; Champion, C.; Château, F.; Chiarusi, T.; Christopoulou, B.; Circella, M.; Classen, L.; Cocimano, R.; Colonges, S.; Coniglione, R.; Cosquer, A.; Costa, M.; Coyle, P.; Creusot, A.; Curtil, C.; Cuttone, G.; D’Amato, C.; D’Amico, A.; De Bonis, G.; De Rosa, G.; Deniskina, N.; Destelle, J. J.; Distefano, C.; Donzaud, C.; Dornic, D.; Dorosti-Hasankiadeh, Q.; Drakopoulou, E.; Drouhin, D.; Drury, L.; Durand, D.; Eberl, T.; Eleftheriadis, C.; Elsaesser, D.; Enzenhöfer, A.; Fermani, P.; Fusco, L. A.; Gajana, D.; Gal, T.; Galatà, S.; Gallo, F.; Garufi, F.; Gebyehu, M.; Giordano, V.; Gizani, N.; Gracia Ruiz, R.; Graf, K.; Grasso, R.; Grella, G.; Grmek, A.; Habel, R.; van Haren, H.; Heid, T.; Heijboer, A.; Heine, E.; Henry, S.; Hernández-Rey, J. J.; Herold, B.; Hevinga, M. A.; van der Hoek, M.; Hofestädt, J.; Hogenbirk, J.; Hugon, C.; Hößl, J.; Imbesi, M.; James, C.; Jansweijer, P.; Jochum, J.; de Jong, M.; Kadler, M.; Kalekin, O.; Kappes, A.; Kappos, E.; Katz, U.; Kavatsyuk, O.; Keller, P.; Kieft, G.; Koffeman, E.; Kok, H.; Kooijman, P.; Koopstra, J.; Korporaal, A.; Kouchner, A.; Koutsoukos, S.; Kreykenbohm, I.; Kulikovskiy, V.; Lahmann, R.; Lamare, P.; Larosa, G.; Lattuada, D.; Le Provost, H.; Leisos, A.; Lenis, D.; Leonora, E.; Lindsey Clark, M.; Liolios, A.; Llorens Alvarez, C. D.; Löhner, H.; Lo Presti, D.; Louis, F.; Maccioni, E.; Mannheim, K.; Manolopoulos, K.; Margiotta, A.; Mariş, O.; Markou, C.; Martínez-Mora, J. A.; Martini, A.; Masullo, R.; Michael, T.; Migliozzi, P.; Migneco, E.; Miraglia, A.; Mollo, C.; Mongelli, M.; Morganti, M.; Mos, S.; Moudden, Y.; Musico, P.; Musumeci, M.; Nicolaou, C.; Nicolau, C. A.; Orlando, A.; Orzelli, A.; Papageorgiou, K.; Papaikonomou, A.; Papaleo, R.; Păvălaş, G. E.; Peek, H.; Pellegrino, C.; Pellegriti, M. G.; Perrina, C.; Petridou, C.; Piattelli, P.; Pikounis, K.; Popa, V.; Pradier, Th; Priede, M.; Pühlhofer, G.; Pulvirenti, S.; Racca, C.; Raffaelli, F.; Randazzo, N.; Rapidis, P. A.; Razis, P.; Real, D.; Resvanis, L.; Reubelt, J.; Riccobene, G.; Rovelli, A.; Royon, J.; Saldaña, M.; Samtleben, D. F E; Sanguineti, M.; Santangelo, A.; Sapienza, P.; Savvidis, I.; Schmelling, J.; Schnabel, J.; Sedita, M.; Seitz, T.; Sgura, I.; Simeone, F.; Siotis, I.; Sipala, V.; Solazzo, M.; Spitaleri, A.; Spurio, M.; Stavropoulos, G.; Steijger, J.; Stolarczyk, T.; Stransky, D.; Taiuti, M.; Terreni, G.; Tézier, D.; Théraube, S.; Thompson, L. F.; Timmer, P.; Trapierakis, H. I.; Trasatti, L.; Trovato, A.; Tselengidou, M.; Tsirigotis, A.; Tzamarias, S.; Tzamariudaki, E.; Vallage, B.; Van Elewyck, V.; Vermeulen, J.; Vernin, P.; Viola, S.; Vivolo, D.; Werneke, P.; Wiggers, L.; Wilms, J.; de Wolf, E.; van Wooning, R. H L; Yatkin, K.; Zachariadou, K.; Zonca, E.; Zornoza, J. D.; Zúñiga, J.; Zwart, A.

    2014-01-01

    The first prototype of a photo-detection unit of the future KM3NeT neutrino telescope has been deployed in the deep waters of the Mediterranean Sea. This digital optical module has a novel design with a very large photocathode area segmented by the use of 31 three inch photomultiplier tubes. It has

  18. NAD(P)H oxidase/nitric oxide interactions in peroxisome proliferator activated receptor (PPAR){alpha}-mediated cardiovascular effects

    Energy Technology Data Exchange (ETDEWEB)

    Newaz, Mohammad [Center for Cardiovascular Diseases, College of Pharmacy and Health Sciences, Texas Southern University, Houston, TX 77004 (United States); Blanton, Ahmad [Center for Cardiovascular Diseases, College of Pharmacy and Health Sciences, Texas Southern University, Houston, TX 77004 (United States); Fidelis, Paul [Center for Cardiovascular Diseases, College of Pharmacy and Health Sciences, Texas Southern University, Houston, TX 77004 (United States); Oyekan, Adebayo [Center for Cardiovascular Diseases, College of Pharmacy and Health Sciences, Texas Southern University, Houston, TX 77004 (United States)]. E-mail: Oyekan_AO@TSU.EDU

    2005-11-11

    Activation of peroxisome proliferator activated receptor (PPAR){alpha} and its protective role in cardiovascular function has been reported but the exact mechanism(s) involved is not clear. As we have shown that PPAR{alpha} ligands increased nitric oxide (NO) production and cardiovascular function is controlled by a balance between NO and free radicals, we hypothesize that PPAR{alpha} activation tilts the balance between NO and free radicals and that this mechanism defines the protective effects of PPAR{alpha} ligands on cardiovascular system. Systolic blood pressure (SBP) was greater in PPAR{alpha} knockout (KO) mice compared with its wild type (WT) litter mates (130 {+-} 10 mmHg versus 107 {+-} 4 mmHg). L-NAME (100 mg/L p.o.), the inhibitor of NO production abolished the difference between PPAR{alpha} KO and WT mice. In kidney homogenates, tissue lipid hydroperoxide generation was greater in KO mice (11.8 {+-} 1.4 pM/mg versus 8.3 {+-} 0.6 pM/mg protein). This was accompanied by a higher total NOS activity (46 {+-} 6%, p < 0.05) and a {approx}3 fold greater Ca{sup 2+}-dependent NOS activity in kidney homogenates of untreated PPAR{alpha} WT compared with the KO mice. Clofibrate, a PPAR{alpha} ligand, increased NOS activity in WT but not KO mice. Bezafibrate (30 mg/kg) reduced SBP in conscious rats (19 {+-} 4%, p < 0.05), increased urinary NO excretion (4.06 {+-} 0.53-7.07 {+-} 1.59 {mu}M/24 h; p < 0.05) and reduced plasma 8-isoprostane level (45.8 {+-} 15 {mu}M versus 31.4 {+-} 8 {mu}M), and NADP(H) oxidase activity (16 {+-} 5%). Implantation of DOCA pellet (20 mg s.c.) in uninephrectomized mice placed on 1% NaCl drinking water increased SBP by a margin that was markedly greater in KO mice (193 {+-} 13 mmHg versus 130 {+-} 12 mmHg). In the rat, DOCA increased SBP and NAD(P)H oxidase activity and both effects were diminished by clofibrate. In addition, clofibrate reduced ET-1 production in DOCA/salt hypertensive rats. Thus, apart from inhibition of ET-1 production

  19. Nmnat1-Rbp7 Is a Conserved Fusion-Protein That Combines NAD+ Catalysis of Nmnat1 with Subcellular Localization of Rbp7.

    Directory of Open Access Journals (Sweden)

    Hao Chen

    Full Text Available Retinol binding proteins (Rbps are known as carriers for transport and targeting of retinoids to their metabolizing enzymes. Rbps are also reported to function in regulating the homeostatic balance of retinoid metabolism, as their level of retinoid occupancy impacts the activities of retinoid metabolizing enzymes. Here we used zebrafish as a model to study rbp7a function and regulation. We find that early embryonic rbp7a expression is negatively regulated by the Nodal/FoxH1-signaling pathway and we show that Nodal/FoxH1 activity has the opposite effect on aldh1a2, which encodes the major enzyme for early embryonic retinoic acid production. The data are consistent with a Nodal-dependent coordination of the allocation of retinoid precursors to processing enzymes with the catalysis of retinoic acid formation. Further, we describe a novel nmnat1-rbp7 transcript encoding a fusion of Rbp7 and the NAD+ (Nicotinamide adenine dinucleotide synthesizing enzyme Nmnat1. We show that nmnat1-rbp7 is conserved in fish, mouse and chicken, and that in zebrafish regulation of nmnat1-rbp7a is distinct from that of rbp7a and nmnat1. Injection experiments in zebrafish further revealed that Nmnat1-Rbp7a and Nmnat1 have similar NAD+ catalyzing activities but a different subcellular localization. HPLC measurements and protein localization analysis highlight Nmnat1-Rbp7a as the only known cytoplasmic and presumably endoplasmic reticulum (ER specific NAD+ catalyzing enzyme. These studies, taken together with previously documented NAD+ dependent interaction of RBPs with ER-associated enzymes of retinal catalysis, implicate functions of this newly described NMNAT1-Rbp7 fusion protein in retinol oxidation.

  20. An Examination by Site-Directed Mutagenesis of Putative Key Residues in the Determination of Coenzyme Specificity in Clostridial NAD+-Dependent Glutamate Dehydrogenase

    OpenAIRE

    Griffin, Joanna; Engel, Paul C.

    2011-01-01

    Sequence and structure comparisons of various glutamate dehydrogenases (GDH) and other nicotinamide nucleotide-dependent dehydrogenases have potentially implicated certain residues in coenzyme binding and discrimination. We have mutated key residues in Clostridium symbiosum NAD+-specific GDH to investigate their contribution to specificity and to enhance acceptance of NADPH. Comparisons with E. coli NADPH-dependent GDH prompted design of mutants F238S, P262S, and F238S/P262S, which were purif...