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Sample records for acetoacetate decarboxylase adc

  1. Kinetic challenges facing oxalate, malonate, acetoacetate, and oxaloacetate decarboxylases.

    PubMed

    Wolfenden, Richard; Lewis, Charles A; Yuan, Yang

    2011-04-20

    To compare the powers of the corresponding enzymes as catalysts, the rates of uncatalyzed decarboxylation of several aliphatic acids (oxalate, malonate, acetoacetate, and oxaloacetate) were determined at elevated temperatures and extrapolated to 25 °C. In the extreme case of oxalate, the rate of the uncatalyzed reaction at pH 4.2 was 1.1 × 10(-12) s(-1), implying a 2.5 × 10(13)-fold rate enhancement by oxalate decarboxylase. Whereas the enzymatic decarboxylation of oxalate requires O(2) and Mn(II), the uncatalyzed reaction is unaffected by the presence of these cofactors and appears to proceed by heterolytic elimination of CO(2).

  2. Hetero- and homodimerization of Arabidopsis thaliana arginine decarboxylase AtADC1 and AtADC2.

    PubMed

    Maruri-López, Israel; Jiménez-Bremont, Juan F

    2017-03-11

    The arginine decarboxylase enzyme (ADC) carries out the production of agmatine from arginine, which is the precursor of the first polyamine (PA) known as putrescine; subsequently, putrescine is turned into the higher PAs, spermidine and spermine. In Arabidopsis thaliana PA production occurs only from arginine and this step is initiated by two ADC paralogues, AtADC1 and AtADC2. PA production is essential for A. thaliana life cycle. Here, we analyzed the sub-cellular localization of AtADC1 and AtADC2 enzymes through GFP translational fusions. Our data revealed that the A. thaliana arginine decarboxylase enzymes exhibit a dual sub-cellular localization both in the cytosol and chloroplast. Moreover, we examined the protein dimer assembly using a Bimolecular Fluorescence Complementation (BiFC) approach, which showed that AtADC1 and AtADC2 proteins were able to form homodimers in the cytosol and chloroplast. Interestingly, we found the formation of AtADC1/AtADC2 heterodimers with similar sub-cellular localization than homodimers. This study reveals that both ADC proteins are located in the same cell compartments, and they are able to form protein interaction complexes with each other.

  3. Role of Arginine decarboxylase (ADC) in Arabidopsis thaliana defence against the pathogenic bacterium Pseudomonas viridiflava.

    PubMed

    Rossi, F R; Marina, M; Pieckenstain, F L

    2015-07-01

    Polyamine biosynthesis starts with putrescine production through the decarboxylation of arginine or ornithine. In Arabidopsis thaliana, putrescine is synthesised exclusively by arginine decarboxylase (ADC), which exists as two isoforms (ADC1 and 2) that are differentially regulated by abiotic stimuli, but their role in defence against pathogens has not been studied in depth. This work analysed the participation of ADC in Arabidopsis defence against Pseudomonas viridiflava. ADC activity and expression, polyamine levels and bacterial resistance were analysed in null mutants of each ADC isoform. In non-infected wild-type (WT) plants, ADC2 expression was much higher than ADC1. Analysis of adc mutants demonstrated that ADC2 contributes to a much higher extent than ADC1 to basal ADC activity and putrescine biosynthesis. In addition, adc2 mutants showed increased basal expression of salicylic acid- and jasmonic acid-dependent PR genes. Bacterial infection induced putrescine accumulation and ADC1 expression in WT plants, but pathogen-induced putrescine accumulation was blocked in adc1 mutants. Results suggest a specific participation of ADC1 in defence, although basal resistance was not decreased by dysfunction of either of the two ADC genes. In addition, and as opposed to WT plants, bacterial infection increased ADC2 expression and ADC activity in adc1 mutants, which could counterbalance the lack of ADC1. Results demonstrate a major contribution of ADC2 to total ADC activity and the specific induction of ADC1 in response to infection. A certain degree of functional redundancy between the two isoforms in relation to their contribution to basal resistance is also evident.

  4. Arginine decarboxylase (ADC) and agmatinase (AGMAT): an alternative pathway for synthesis of polyamines in pig conceptuses and uteri

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Arginine, a precursor for the synthesis of nitric oxide (NO) and polyamines, is critical for implantation and development of the conceptus. We first reported that the arginine decarboxylase (ADC)/agmatinase(AGMAT) pathway as an alternative pathway for synthesis of polyamines in the ovine conceptuses...

  5. An arginine decarboxylase gene PtADC from Poncirus trifoliata confers abiotic stress tolerance and promotes primary root growth in Arabidopsis.

    PubMed

    Wang, Jing; Sun, Pei-Pei; Chen, Chun-Li; Wang, Yin; Fu, Xing-Zheng; Liu, Ji-Hong

    2011-05-01

    Arginine decarboxylase (ADC) is an important enzyme responsible for polyamine synthesis under stress conditions. In this study, the gene encoding ADC in Poncirus trifoliata (PtADC) was isolated and it existed as a single-copy member. Transcript levels of PtADC were up-regulated by low temperature and dehydration. Overexpression of PtADC in an Arabidopsis thaliana ADC mutant adc1-1 promoted putrescine synthesis in the transgenic line and the stomatal density was reverted to that in the wild type. The transgenic line showed enhanced resistance to high osmoticum, dehydration, long-term drought, and cold stress compared with the wild type and the mutant. The accumulation of reactive oxygen species (ROS) in the transgenic line was appreciably decreased under the stresses, but ROS scavenging capacity was compromised when the transgenic plants were treated with the ADC inhibitor D-arginine prior to stress treatment. In addition, the transgenic line had longer roots than the wild type and the mutant under both normal and stressful conditions, consistent with larger cell number and length of the root meristematic zone. Taken together, these results demonstrated that PtADC is involved in tolerance to multiple stresses, and its function may be due, at least partly, to efficient ROS elimination and to its influence on root growth conducive to drought tolerance.

  6. Acetone production in solventogenic Clostridium species: new insights from non-enzymatic decarboxylation of acetoacetate.

    PubMed

    Han, Bei; Gopalan, Venkat; Ezeji, Thaddeus Chukwuemeka

    2011-08-01

    Development of a butanologenic strain with high selectivity for butanol production is often proposed as a possible route for improving the economics of biobutanol production by solventogenic Clostridium species. The acetoacetate decarboxylase (aadc) gene encoding acetoacetate decarboxylase (AADC), which catalyzes the decarboxylation of acetoacetate into acetone and CO(2), was successfully disrupted by homologous recombination in solventogenic Clostridium beijerinckii NCIMB 8052 to generate an aadc ( - ) mutant. Our fermentation studies revealed that this mutant produces a maximum acetone concentration of 3 g/L (in P2 medium), a value comparable to that produced by wild-type C. beijerinckii 8052. Therefore, we postulated that AADC-catalyzed decarboxylation of acetoacetate is not the sole means for acetone generation. Our subsequent finding that non-enzymatic decarboxylation of acetoacetate in vitro, under conditions similar to in vivo acetone-butanol-ethanol (ABE) fermentation, produces 1.3 to 5.2 g/L acetone between pH 6.5 and 4 helps rationalize why various knock-out and knock-down strategies designed to disrupt aadc in solventogenic Clostridium species did not eliminate acetone production during ABE fermentation. Based on these results, we discuss alternatives to enhance selectivity for butanol production.

  7. Arginine Decarboxylase Is Localized in Chloroplasts.

    PubMed Central

    Borrell, A.; Culianez-Macia, F. A.; Altabella, T.; Besford, R. T.; Flores, D.; Tiburcio, A. F.

    1995-01-01

    Plants, unlike animals, can use either ornithine decarboxylase or arginine decarboxylase (ADC) to produce the polyamine precursor putrescine. Lack of knowledge of the exact cellular and subcellular location of these enzymes has been one of the main obstacles to our understanding of the biological role of polyamines in plants. We have generated polyclonal antibodies to oat (Avena sativa L.) ADC to study the spatial distribution and subcellular localization of ADC protein in different oat tissues. By immunoblotting and immunocytochemistry, we show that ADC is organ specific. By cell fractionation and immunoblotting, we show that ADC is localized in chloroplasts associated with the thylakoid membrane. The results also show that increased levels of ADC protein are correlated with high levels of ADC activity and putrescine in osmotically stressed oat leaves. A model of compartmentalization for the arginine pathway and putrescine biosynthesis in active photosynthetic tissues has been proposed. In the context of endosymbiote-driven metabolic evolution in plants, the location of ADC in the chloroplast compartment may have major evolutionary significance, since it explains (a) why plants can use two alternative pathways for putrescine biosynthesis and (b) why animals do not possess ADC. PMID:12228631

  8. Localization of arginine decarboxylase in tobacco plants.

    PubMed

    Bortolotti, Cristina; Cordeiro, Alexandra; Alcázar, Rubén; Borrell, Antoni; Culiañez-Macià, Francisco A.; Tiburcio, Antonio F.; Altabella, Teresa

    2004-01-01

    The lack of knowledge about the tissue and subcellular distribution of polyamines (PAs) and the enzymes involved in their metabolism remains one of the main obstacles in our understanding of the biological role of PAs in plants. Arginine decarboxylase (ADC; EC 4.1.1.9) is a key enzyme in polyamine biosynthesis in plants. We have characterized a cDNA coding for ADC from Nicotiana tabacum L. cv. Petit Havana SR1. The deduced ADC polypeptide had 721 amino acids and a molecular mass of 77 kDa. The ADC cDNA was overexpressed in Escherichia coli, and the ADC fusion protein obtained was used to produce polyclonal antibodies. Using immunological methods, we demonstrate the presence of the ADC protein in all plant organs analysed: flowers, seeds, stems, leaves and roots. Moreover, depending on the tissue, the protein is localized in two different subcellular compartments, the nucleus and the chloroplast. In photosynthetic tissues, ADC is located mainly in chloroplasts, whereas in non-photosynthetic tissues the protein appears to be located in nuclei. The different compartmentation of ADC may be related to distinct functions of the protein in different cell types.

  9. Arginine decarboxylase as the source of putrescine for tobacco alkaloids

    NASA Technical Reports Server (NTRS)

    Tiburcio, A. F.; Galston, A. W.

    1986-01-01

    The putrescine which forms a part of nicotine and other pyrrolidine alkaloids is generally assumed to arise through the action of ornithine decarboxylase (ODC). However, we have previously noted that changes in the activity of arginine decarboxylase (ADC), an alternate source of putrescine, parallel changes in tissue alkaloids, while changes in ODC activity do not. This led us to undertake experiments to permit discrimination between ADC and ODC as enzymatic sources of putrescine destined for alkaloids. Two kinds of evidence presented here support a major role for ADC in the generation of putrescine going into alkaloids: (a) A specific 'suicide inhibitor' of ADC effectively inhibits the biosynthesis of nicotine and nornicotine in tobacco callus, while the analogous inhibitor of ODC is less effective, and (b) the flow of 14C from uniformly labelled arginine into nicotine is much more efficient than that from ornithine.

  10. Molecular and functional analyses of amino acid decarboxylases involved in cuticle tanning in Tribolium castaneum

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aspartate 1-decarboxylase (ADC) and dopa decarboxylase (DDC) provide b–alanine and dopamine used in insect cuticle tanning. Beta-alanine is conjugated with dopamine to yield N-b-alanyldopamine (NBAD), a substrate for the phenoloxidase laccase that catalyzes the synthesis of cuticle protein cross-li...

  11. Expression of arginine decarboxylase in brain regions and neuronal cells

    PubMed Central

    Iyo, Abiye H.; Zhu, Meng-Yang; Ordway, Gregory A.; Regunathan, Soundar

    2010-01-01

    After our initial report of a mammalian gene for arginine decarboxylase, an enzyme for the synthesis of agmatine from arginine, we have determined the regional expression of ADC in rat. We have analyzed the expression of ADC in rat brain regions by activity, protein and mRNA levels, and the regulation of expression in neuronal cells by RNA interference. In rat brain, ADC was widely expressed in major brain regions, with a substantial amount in hypothalamus, followed by cortex, and with least amounts in locus coeruleus and medulla. ADC mRNA was detected in primary astrocytes and C6 glioma cells. While no ADC message was detected in fresh neurons (3 days old), significant message appeared in differentiated neurons (3 weeks old). PC12 cells, treated with nerve growth factor, had higher ADC mRNA compared with naive cells. The siRNA mixture directed towards the N-terminal regions of ADC cDNA down-regulated the levels of mRNA and protein in cultured neurons/C6 glioma cells and these cells produced lower agmatine. Thus, this study demonstrates that ADC message is expressed in rat brain regions, that it is regulated in neuronal cells and that the down-regulation of ADC activity by specific siRNA leads to lower agmatine production. PMID:16445852

  12. Simultaneous Silencing of Two Arginine Decarboxylase Genes Alters Development in Arabidopsis

    PubMed Central

    Sánchez-Rangel, Diana; Chávez-Martínez, Ana I.; Rodríguez-Hernández, Aída A.; Maruri-López, Israel; Urano, Kaoru; Shinozaki, Kazuo; Jiménez-Bremont, Juan F.

    2016-01-01

    Polyamines (PAs) are small aliphatic polycations that are found ubiquitously in all organisms. In plants, PAs are involved in diverse biological processes such as growth, development, and stress responses. In Arabidopsis thaliana, the arginine decarboxylase enzymes (ADC1 and 2) catalyze the first step of PA biosynthesis. For a better understanding of PA biological functions, mutants in PA biosynthesis have been generated; however, the double adc1/adc2 mutant is not viable in A. thaliana. In this study, we generated non-lethal A. thaliana lines through an artificial microRNA that simultaneously silenced the two ADC genes (amiR:ADC). The generated transgenic lines (amiR:ADC-L1 and -L2) showed reduced AtADC1 and AtADC2 transcript levels. For further analyses the amiR:ADC-L2 line was selected. We found that the amiR:ADC-L2 line showed a significant decrease of their PA levels. The co-silencing revealed a stunted growth in A. thaliana seedlings, plantlets and delay in its flowering rate; these phenotypes were reverted with PA treatment. In addition, amiR:ADC-L2 plants displayed two seed phenotypes, such as yellow and brownish seeds. The yellow mutant seeds were smaller than adc1, adc2 mutants and wild type seeds; however, the brownish were the smallest seeds with arrested embryos at the torpedo stage. These data reinforce the importance of PA homeostasis in the plant development processes. PMID:27014322

  13. Deuterium Exchange in Ethyl Acetoacetate: An Undergraduate GC-MS [Gas Chromatography-Mass Spectroscopy] Experiment

    ERIC Educational Resources Information Center

    Heinson, C. D.; Williams, J. M.; Tinnerman, W. N.; Malloy, T. B.

    2005-01-01

    The role of ethanol O-d in nullifying the deuterolysis may be demonstrated by determining that transesterification of methyl acetoacetate of the ethyl ester occurs as well as deuterium exchange of the five acetoacetate hydrogens. The significant acidity of the methylene protons in the acetoacetate group, the efficacy of base catalysis, the role of…

  14. Lipase-catalyzed synthesis of partial acylglycerols of acetoacetate

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A commercially available immobilized preparation of Rhizomucor miehei lipase (Lipozyme RMIM) has been employed in the synthesis of partial glycerides of acetoacetate. Due to the chemical reactivitity of the acetoacetyl group, these glycerides could have novel uses in e.g. polymer formation. Both 1...

  15. Whole cells in enantioselective reduction of benzyl acetoacetate

    PubMed Central

    Ribeiro, Joyce Benzaquem; Ramos, Aline de Souza; Lopes, Raquel de Oliveira; da Silva, Gabriela Veloso Vieira; de Souza, Rodrigo Octavio Mendonça Alves

    2014-01-01

    The β-ketoester benzyl acetoacetate was enantioselectively reduced to benzyl (S)-3-hydroxybutanoate by seven microorganism species. The best result using free cells was obtained with the yeast Hansenula sp., which furnished 97% ee and 85% of conversion within 24 h. After immobilization in calcium alginate spheres, K.marxianus showed to be more stable after 2 cycles of reaction. PMID:25477927

  16. Molecular cloning and expression analysis of an arginine decarboxylase gene from peach (Prunus persica).

    PubMed

    Liu, Ji Hong; Ban, Yusuke; Wen, Xiao-Peng; Nakajima, Ikuko; Moriguchi, Takaya

    2009-01-15

    Arginine decarboxylase (ADC), one of the enzymes responsible for putrescine (Put) biosynthesis, has been shown to be implicated in stress response. In the current paper attempts were made to clone and characterize a gene encoding ADC from peach (Prunus persica (L.) Batsch, 'Akatsuki'). Rapid amplification of cDNA ends (RACE) gave rise to a full-length ADC cDNA (PpADC) with a complete open reading frame of 2178 bp, encoding a 725 amino acid polypeptide. Homology search and sequence multi-alignment demonstrated that the deduced PpADC protein sequence shared a high identity with ADCs from other plants, including several highly conservative motifs and amino acids. Southern blotting indicated that PpADC existed in peach genome as a single gene. Expression levels of PpADC in different tissues of peach (P. persica 'Akatsuki') were spatially and developmentally regulated. Treatment of peach shoots from 'Mochizuki' with exogenous 5 mM Put, an indirect product of ADC, remarkably induced accumulation of PpADC mRNA. Transcripts of PpADC in peach leaves from 'Mochizuki' were quickly induced, either transiently or continuously, in response to dehydration, high salinity (200 mM NaCl), low temperature (4 degrees C) and heavy metal (150 microM CdCl(2)), but repressed by high temperature 37 degrees C) during a 2-day treatment, which changed in an opposite direction when the stresses were otherwise removed with the exception of CdCl(2) treatment. In addition, steady-state of PpADC mRNA could be also transiently up-regulated by abscisic acid (ABA) in 'Mochizuki' leaves. All of these, taken together, suggest that PpADC is a stress-responsive gene and can be considered as a potential target that is genetically manipulated so as to create novel germplasms with enhanced stress tolerance in the future.

  17. /sup 3/H-DFMA metabolism in tobacco: non-specific, arginase mediated inhibition of ornithine decarboxylase activity

    SciTech Connect

    Slocum, R.D.; Feirer, R.L.

    1987-04-01

    ..cap alpha..-Difluoromethylarginine (DFMA) is a specific, enzyme-activated, irreversible inhibit of arginine decarboxylase (ADC) in vitro. ADC catalyzes the first step leading to putrescine biosynthesis and the activity of this enzyme is closely linked to overall polyamine (PA) biosynthesis in non-meristematic plant tissues. Consequently, ADC represents an important target enzyme for inhibitors of PA metabolism. DFMA has been shown to inhibit ADC activity in a variety of tissues in vivo but its specificity in tobacco was questioned since ornithine decarboxylase (ODC) activity was also inhibited. Further studies have shown that (/sup 3/H)-DFMA is efficiently hydrolyzed in tobacco to (/sup 3/H)-difluoromethylornithine (DFMO), an irreversible inhibitor of ODC. Tobacco and bovine arginases also catalyze the hydrolysis of DFMA in vitro, suggesting a role for this enzyme in mediating the non-specific inhibition of ODC by DFMA in tobacco flowers.

  18. Arginine and Ornithine Decarboxylases, the Polyamine Biosynthetic Enzymes of Mung Bean Seedlings 1

    PubMed Central

    Altman, Arie; Friedman, Ra'Anan; Levin, Nitsa

    1982-01-01

    General properties and relative activities of l-arginine decarboxylase (ADC) (EC 4.1.1.19) and l-ornithine decarboxylase (ODC) (EC 4.1.1.17), two important enzymes in putrescine and polyamine biosynthesis, were investigated in mung bean (Vigna radiata L.) tissues. Both activities increase linearly with increasing concentrations of crude enzyme, but the increase in ADC activity is considerably greater. The decarboxylation reaction is linear for up to 30 to 60 minutes, and both enzymes have a pH optimum of 7.2. α-Difluoromethyl-ornithine inhibits ODC activity of excised roots, while increasing ADC activity. High specific activity of both enzymes is detected in terminal buds and leaves, while root and hypocotyl activity is low. Different ADC-to-ODC activity ratios are found in various tissues of mung bean plants. Substantial increase in the activity of both enzymes is detected in incubated sections as compared with intact plants. A comparison of several plant species indicates a wide range of ADC-to-ODC activity ratio. It is suggested that both ADC and ODC are active in plant tissues and that their relative contribution to putrescine biosynthesis is dependent upon the type of tissue and growth process. PMID:16662312

  19. Dynamic Testing of ADC: A Review

    NASA Astrophysics Data System (ADS)

    Garg, Bhawana; Mishra, D. K.

    2012-09-01

    Analog to digital converters are Mixed signal devices. With the increasing popularity of these devices , it is important to get more faster and accurate device. Along with design, testing of ADC plays major role . Static and Dynamic methods are available for ADC testing. Ideal ADC itself has quantization error. A NonIdeal ADC consists many other errors like offset error, Gain error, DNL, INL,ENOB,SNR,THD,SINAD etc. In this paper various methods of dynamic testing of ADC are discussed.

  20. PET study of 11C-acetoacetate kinetics in rat brain during dietary treatments affecting ketosis.

    PubMed

    Bentourkia, M'hamed; Tremblay, Sébastien; Pifferi, Fabien; Rousseau, Jacques; Lecomte, Roger; Cunnane, Stephen

    2009-04-01

    Normally, the brain's fuel is glucose, but during fasting it increasingly relies on ketones (beta-hydroxybutyrate, acetoacetate, and acetone) produced in liver mitochondria from fatty acid beta-oxidation. Although moderately raised blood ketones produced on a very high fat ketogenic diet have important clinical effects on the brain, including reducing seizures, ketone metabolism by the brain is still poorly understood. The aim of the present work was to assess brain uptake of carbon-11-labeled acetoacetate (11C-acetoacetate) by positron emission tomography (PET) imaging in the intact, living rat. To vary plasma ketones, we used three dietary conditions: high carbohydrate control diet (low plasma ketones), fat-rich ketogenic diet (raised plasma ketones), and 48-h fasting (raised plasma ketones). 11C-acetoacetate metabolism was measured in the brain, heart, and tissue in the mouth area. Using 11C-acetoacetate and small animal PET imaging, we have noninvasively quantified an approximately seven- to eightfold enhanced brain uptake of ketones on a ketogenic diet or during fasting. This opens up an opportunity to study brain ketone metabolism in humans.

  1. Changes in citric acid cycle flux and anaplerosis antedate the functional decline in isolated rat hearts utilizing acetoacetate.

    PubMed Central

    Russell, R R; Taegtmeyer, H

    1991-01-01

    To determine the temporal relationship between changes in contractile performance and flux through the citric acid cycle in hearts oxidizing acetoacetate, we perfused isolated working rat hearts with either glucose or acetoacetate (both 5 mM) and freeze-clamped the tissue at defined times. After 60 min of perfusion, hearts utilizing acetoacetate exhibited lower systolic and diastolic pressures and lower cardiac outputs. The oxidation of acetoacetate increased the tissue content of 2-oxoglutarate and glutamate and decreased the content of succinyl-CoA suggesting inhibition of citric acid cycle flux through 2-oxoglutarate dehydrogenase. Whereas hearts perfused with either acetoacetate or glucose were similar with respect to their function for the first 20 min, changes in tissue metabolites were already observed within 5 min of perfusion at near-physiological workloads. The addition of lactate or propionate, but not acetate, to hearts oxidizing acetoacetate improved contractile performance, although inhibition of 2-oxoglutarate dehydrogenase was probably not diminished. If lactate or propionate were added, malate and citrate accumulated indicating utilization of anaplerotic pathways for the citric acid cycle. We conclude that a decreased rate of flux through 2-oxoglutarate dehydrogenase in hearts oxidizing acetoacetate precedes, and may be responsible for, contractile failure and is not the result of decreased cardiac work. Further, anaplerosis play an important role in the maintenance of contractile function in hearts utilizing acetoacetate. Images PMID:1671390

  2. Aspartate Decarboxylase is Required for a Normal Pupa Pigmentation Pattern in the Silkworm, Bombyx mori

    PubMed Central

    Dai, Fangyin; Qiao, Liang; Cao, Cun; Liu, Xiaofan; Tong, Xiaoling; He, Songzhen; Hu, Hai; Zhang, Li; Wu, Songyuan; Tan, Duan; Xiang, Zhonghuai; Lu, Cheng

    2015-01-01

    The pigmentation pattern of Lepidoptera varies greatly in different development stages. To date, the effects of key genes in the melanin metabolism pathway on larval and adult body color are distinct, yet the effects on pupal pigmentation remains unclear. In the silkworm, Bombyx mori, the black pupa (bp) mutant is only specifically melanized at the pupal stage. Using positional cloning, we found that a mutation in the Aspartate decarboxylase gene (BmADC) is causative in the bp mutant. In the bp mutant, a SINE-like transposon with a length of 493 bp was detected ~2.2 kb upstream of the transcriptional start site of BmADC. This insertion causes a sharp reduction in BmADC transcript levels in bp mutants, leading to deficiency of β-alanine and N-β-alanyl dopamine (NBAD), but accumulation of dopamine. Following injection of β-alanine into bp mutants, the color pattern was reverted that of the wild-type silkworms. Additionally, melanic pupae resulting from knock-down of BmADC in the wild-type strain were obtained. These findings show that BmADC plays a crucial role in melanin metabolism and in the pigmentation pattern of the silkworm pupal stage. Finally, this study contributes to a better understanding of pupa pigmentation patterns in Lepidoptera. PMID:26077025

  3. Aspartate Decarboxylase is Required for a Normal Pupa Pigmentation Pattern in the Silkworm, Bombyx mori.

    PubMed

    Dai, Fangyin; Qiao, Liang; Cao, Cun; Liu, Xiaofan; Tong, Xiaoling; He, Songzhen; Hu, Hai; Zhang, Li; Wu, Songyuan; Tan, Duan; Xiang, Zhonghuai; Lu, Cheng

    2015-06-16

    The pigmentation pattern of Lepidoptera varies greatly in different development stages. To date, the effects of key genes in the melanin metabolism pathway on larval and adult body color are distinct, yet the effects on pupal pigmentation remains unclear. In the silkworm, Bombyx mori, the black pupa (bp) mutant is only specifically melanized at the pupal stage. Using positional cloning, we found that a mutation in the Aspartate decarboxylase gene (BmADC) is causative in the bp mutant. In the bp mutant, a SINE-like transposon with a length of 493 bp was detected ~2.2 kb upstream of the transcriptional start site of BmADC. This insertion causes a sharp reduction in BmADC transcript levels in bp mutants, leading to deficiency of β-alanine and N-β-alanyl dopamine (NBAD), but accumulation of dopamine. Following injection of β-alanine into bp mutants, the color pattern was reverted that of the wild-type silkworms. Additionally, melanic pupae resulting from knock-down of BmADC in the wild-type strain were obtained. These findings show that BmADC plays a crucial role in melanin metabolism and in the pigmentation pattern of the silkworm pupal stage. Finally, this study contributes to a better understanding of pupa pigmentation patterns in Lepidoptera.

  4. Molecular cloning and functional identification of a plant ornithine decarboxylase cDNA.

    PubMed

    Michael, A J; Furze, J M; Rhodes, M J; Burtin, D

    1996-02-15

    A cDNA for a plant ornithine decarboxylase (ODC), a key enzyme in putrescine and polyamine biosynthesis, has been isolated from root cultures of the solanaceous plant Datura stramonium. Reverse transcription-PCR employing degenerate oligonucleotide primers representing conserved motifs from other eukaryotic ODCs was used to isolate the cDNA. The longest open reading frame potentially encodes a peptide of 431 amino acids and exhibits similarity to other eukaryotic ODCs, prokaryotic and eukaryotic arginine decarboxylases (ADCs), prokaryotic meso-diaminopimelate decarboxylases and the product of the tabA gene of Pseudomonas syringae cv. tabaci. Residues involved at the active site of the mouse ODC are conserved in the plant enzyme. The plant ODC does not possess the C-terminal extension found in the mammalian enzyme, implicated in rapid turnover of the protein, suggesting that the plant ODC may have a longer half-life. Expression of the plant ODC in Escherichia coli and demonstration of ODC activity confirmed that the cDNA encodes an active ODC enzyme. This is the first description of the primary structure of a eukaryotic ODC isolated from an organism where the alternative ADC routine to putrescine is present.

  5. Arginine Decarboxylase expression, polyamines biosynthesis and reactive oxygen species during organogenic nodule formation in hop.

    PubMed

    Fortes, Ana M; Costa, Joana; Santos, Filipa; Seguí-Simarro, José M; Palme, Klaus; Altabella, Teresa; Tiburcio, Antonio F; Pais, Maria S

    2011-02-01

    Hop (Humulus lupulus L.) is an economically important plant species used in beer production and as a health-promoting medicine. Hop internodes develop upon stress treatments organogenic nodules which can be used for genetic transformation and micropropagation. Polyamines are involved in plant development and stress responses. Arginine decarboxylase (ADC; EC 4·1.1·19) is a key enzyme involved in the biosynthesis of putrescine in plants. Here we show that ADC protein was increasingly expressed at early stages of hop internode culture (12h). Protein continued accumulating until organogenic nodule formation after 28 days, decreasing thereafter. The same profile was observed for ADC transcript suggesting transcriptional regulation of ADC gene expression during morphogenesis. The highest transcript and protein levels observed after 28 days of culture were accompanied by a peak in putrescine levels. Reactive oxygen species accumulate in nodular tissues probably due to stress inherent to in vitro conditions and enhanced polyamine catabolism. Conjugated polyamines increased during plantlet regeneration from nodules suggesting their involvement in plantlet formation and/or in the control of free polyamine levels. Immunogold labeling revealed that ADC is located in plastids, nucleus and cytoplasm of nodular cells. In vacuolated cells, ADC immunolabelling in plastids doubled the signal of proplastids in meristematic cells. Location of ADC in different subcellular compartments may indicate its role in metabolic pathways taking place in these compartments. Altogether these data suggest that polyamines play an important role in organogenic nodule formation and represent a progress towards understanding the role played by these growth regulators in plant morphogenesis.

  6. Capacitor-Chain Successive-Approximation ADC

    NASA Technical Reports Server (NTRS)

    Cunningham, Thomas

    2003-01-01

    A proposed successive-approximation analog-to-digital converter (ADC) would contain a capacitively terminated chain of identical capacitor cells. Like a conventional successive-approximation ADC containing a bank of binary-scaled capacitors, the proposed ADC would store an input voltage on a sample-and-hold capacitor and would digitize the stored input voltage by finding the closest match between this voltage and a capacitively generated sum of binary fractions of a reference voltage (Vref). However, the proposed capacitor-chain ADC would offer two major advantages over a conventional binary-scaled-capacitor ADC: (1) In a conventional ADC that digitizes to n bits, the largest capacitor (representing the most significant bit) must have 2(exp n-1) times as much capacitance, and hence, approximately 2(exp n-1) times as much area as does the smallest capacitor (representing the least significant bit), so that the total capacitor area must be 2(exp n) times that of the smallest capacitor. In the proposed capacitor-chain ADC, there would be three capacitors per cell, each approximately equal to the smallest capacitor in the conventional ADC, and there would be one cell per bit. Therefore, the total capacitor area would be only about 3(exp n) times that of the smallest capacitor. The net result would be that the proposed ADC could be considerably smaller than the conventional ADC. (2) Because of edge effects, parasitic capacitances, and manufacturing tolerances, it is difficult to make capacitor banks in which the values of capacitance are scaled by powers of 2 to the required precision. In contrast, because all the capacitors in the proposed ADC would be identical, the problem of precise binary scaling would not arise.

  7. Genetics Home Reference: malonyl-CoA decarboxylase deficiency

    MedlinePlus

    ... deficiency of malonyl-CoA decarboxylase malonic aciduria malonyl-coenzyme A decarboxylase deficiency MCD deficiency Related Information How ... molecular characterization of nine new patients with malonyl-coenzyme A decarboxylase deficiency. J Inherit Metab Dis. 2007 ...

  8. Genetics Home Reference: aromatic l-amino acid decarboxylase deficiency

    MedlinePlus

    ... l-amino acid decarboxylase deficiency aromatic l-amino acid decarboxylase deficiency Enable Javascript to view the expand/ ... Open All Close All Description Aromatic l-amino acid decarboxylase (AADC) deficiency is an inherited disorder that ...

  9. An archaeal glutamate decarboxylase homolog functions as an aspartate decarboxylase and is involved in β-alanine and coenzyme A biosynthesis.

    PubMed

    Tomita, Hiroya; Yokooji, Yuusuke; Ishibashi, Takuya; Imanaka, Tadayuki; Atomi, Haruyuki

    2014-03-01

    β-Alanine is a precursor for coenzyme A (CoA) biosynthesis and is a substrate for the bacterial/eukaryotic pantothenate synthetase and archaeal phosphopantothenate synthetase. β-Alanine is synthesized through various enzymes/pathways in bacteria and eukaryotes, including the direct decarboxylation of Asp by aspartate 1-decarboxylase (ADC), the degradation of pyrimidine, or the oxidation of polyamines. However, in most archaea, homologs of these enzymes are not present; thus, the mechanisms of β-alanine biosynthesis remain unclear. Here, we performed a biochemical and genetic study on a glutamate decarboxylase (GAD) homolog encoded by TK1814 from the hyperthermophilic archaeon Thermococcus kodakarensis. GADs are distributed in all three domains of life, generally catalyzing the decarboxylation of Glu to γ-aminobutyrate (GABA). The recombinant TK1814 protein displayed not only GAD activity but also ADC activity using pyridoxal 5'-phosphate as a cofactor. Kinetic studies revealed that the TK1814 protein prefers Asp as its substrate rather than Glu, with nearly a 20-fold difference in catalytic efficiency. Gene disruption of TK1814 resulted in a strain that could not grow in standard medium. Addition of β-alanine, 4'-phosphopantothenate, or CoA complemented the growth defect, whereas GABA could not. Our results provide genetic evidence that TK1814 functions as an ADC in T. kodakarensis, providing the β-alanine necessary for CoA biosynthesis. The results also suggest that the GAD activity of TK1814 is not necessary for growth, at least under the conditions applied in this study. TK1814 homologs are distributed in a wide range of archaea and may be responsible for β-alanine biosynthesis in these organisms.

  10. An Archaeal Glutamate Decarboxylase Homolog Functions as an Aspartate Decarboxylase and Is Involved in β-Alanine and Coenzyme A Biosynthesis

    PubMed Central

    Tomita, Hiroya; Yokooji, Yuusuke; Ishibashi, Takuya; Imanaka, Tadayuki

    2014-01-01

    β-Alanine is a precursor for coenzyme A (CoA) biosynthesis and is a substrate for the bacterial/eukaryotic pantothenate synthetase and archaeal phosphopantothenate synthetase. β-Alanine is synthesized through various enzymes/pathways in bacteria and eukaryotes, including the direct decarboxylation of Asp by aspartate 1-decarboxylase (ADC), the degradation of pyrimidine, or the oxidation of polyamines. However, in most archaea, homologs of these enzymes are not present; thus, the mechanisms of β-alanine biosynthesis remain unclear. Here, we performed a biochemical and genetic study on a glutamate decarboxylase (GAD) homolog encoded by TK1814 from the hyperthermophilic archaeon Thermococcus kodakarensis. GADs are distributed in all three domains of life, generally catalyzing the decarboxylation of Glu to γ-aminobutyrate (GABA). The recombinant TK1814 protein displayed not only GAD activity but also ADC activity using pyridoxal 5′-phosphate as a cofactor. Kinetic studies revealed that the TK1814 protein prefers Asp as its substrate rather than Glu, with nearly a 20-fold difference in catalytic efficiency. Gene disruption of TK1814 resulted in a strain that could not grow in standard medium. Addition of β-alanine, 4′-phosphopantothenate, or CoA complemented the growth defect, whereas GABA could not. Our results provide genetic evidence that TK1814 functions as an ADC in T. kodakarensis, providing the β-alanine necessary for CoA biosynthesis. The results also suggest that the GAD activity of TK1814 is not necessary for growth, at least under the conditions applied in this study. TK1814 homologs are distributed in a wide range of archaea and may be responsible for β-alanine biosynthesis in these organisms. PMID:24415726

  11. Corticosteroids in nocturnal blood plasma of cows in the field related to stage of lactation and plasma acetoacetate.

    PubMed

    Blom, A K; Halse, K

    1975-02-01

    Two materials of nocturnal blood plasma samples, each representing more than 100 cows unselected with respect to health condition, showed acetoacetate variations within a wide range, overlapping with the range for animals with clinical ketosis. Within each of the two materials a statistically significant (P smaller than 0.001) negative correlation (r equal to minus 0.30) was obtained between plasma corticosteroids and log acetoacetate. The lowest corticosteroid and the highest acetoacetate averages were obtained for animals which were within the first month of the lactation cycle. With increasing time post-partum the two components varied inversely, a maximum in corticosteroids coinciding with a minimum in acetoacetate in cows which had calved 2 1/2 to 3 months before the date of sampling. At this stage the mean plasma sugar was about 10 per cent higher than in early lactation. The corticosteroid ranges for different levels of acetoacetate overlapped extensively. Still cows with acetoacetate smaller than or equal to 0.6 mg/100 ml gave significantly higher corticosteroid averages than the rest of the material. But in animals exceeding the 0.6 mg limit no relationship was found between the severity of the ketonaemia and the steroid level. Thus, no difference was obtained between cows with clinical ketosis and animals which were moderately ketonaemic. Some change in adrenal function may be associated with the development of ketonaemic conditions in cows. But no proof has been provided of a direct adrenal involvement in the transformation of a case of subclinical ketosis to the clinical stage.

  12. Acetoacetate promotes the formation of fluorescent advanced glycation end products (AGEs).

    PubMed

    Bohlooli, Mousa; Ghaffari-Moghaddam, Mansour; Khajeh, Mostafa; Aghashiri, Zohre; Sheibani, Nader; Moosavi-Movahedi, Ali Akbar

    2016-12-01

    Acetoacetate (AA) is an important ketone body, which produces reactive oxygen species (ROS). Advanced glycation end products (AGEs) are defined as final products of glycation process whose production is influenced by the levels of ROS. The accumulation of AGEs in the body contributes to pathogenesis of many diseases including complications of diabetes, and Alzheimer's and Parkinson's disease. Here, we evaluated the impact of AA on production of AGEs upon incubation of human serum albumin (HSA) with glucose. The effect of AA on the AGEs formation of HSA was studied under physiological conditions after incubation with glucose for 35 days. The physical techniques including circular dichroism (CD) and fluorescence spectroscopy were used to assess the impact of AA on formation and structural changes of glycated HSA (GHSA). Our results indicated that the secondary and tertiary structural changes of GHSA were increased in the presence of AA. The fluorescence intensity measurements of AGEs also showed an increase in AGEs formation. Acetoacetate has an activator effect in formation of AGEs through ROS production. The presence of AA may result in enhanced glycation in the presence of glucose and severity of complications associated with accumulation of AGEs.

  13. Polyamines induced by osmotic stress protect Synechocystis sp. PCC 6803 cells and arginine decarboxylase transcripts against UV-B radiation.

    PubMed

    Pothipongsa, Apiradee; Jantaro, Saowarath; Incharoensakdi, Aran

    2012-11-01

    The effect of UV-B radiation on growth and polyamines content of Synechocystis sp. PCC 6803 subjected to either NaCl or sorbitol stress was investigated. Cells could not grow in the presence of 350 mM NaCl or 500 mM sorbitol under normal white light. However, cells grown in BG11 under osmotic stress imposed by NaCl or sorbitol followed by ultraviolet-B (UV-B) irradiation for 2 h showed higher cell density than those under the same condition but no osmotic stress. The chlorophyll fluorescence parameter (F(v)/F(m)) also showed an apparent decrease upon UV-B irradiation. Intracellular polyamines increased by about 2- and 4-fold in NaCl- and sorbitol-stressed cells, respectively. When these cells were irradiated with UV-B for 1 h, a further 3-fold increase in polyamines content was detected in NaCl-stressed but not sorbitol-stressed cells. Synechocystis cells contained adc1 and adc2 genes encoding arginine decarboxylase (ADC) with only adc1 showing upregulation by NaCl or sorbitol stress. NaCl- or sorbitol-stressed cells contained about 5-fold higher level of adc1 transcript than did the unstressed cells after 1-h irradiation with UV-B, suggesting the protection of adc1 transcript by accumulated polyamines, due to NaCl or sorbitol stress, against UV-B radiation damage. ADC levels as analyzed by Western blot showed upregulation by UV-B in NaCl-stressed but not sorbitol-stressed cells.

  14. Overexpression of PtADC confers enhanced dehydration and drought tolerance in transgenic tobacco and tomato: effect on ROS elimination.

    PubMed

    Wang, Bao-Quan; Zhang, Qing-Fu; Liu, Ji-Hong; Li, Guo-Huai

    2011-09-16

    Drought is a major environmental factor that limits plant growth and productivity. Polyamines have been shown to act as stress molecules that accumulate in plant adaptation to abiotic stresses. In this study, an arginine decarboxylase gene isolated from Poncirus trifoliata, PtADC, was introduced into tobacco and tomato to investigate its function in drought tolerance. We demonstrate that the transgenic plants showed an improvement in dehydration and drought tolerance. Under dehydration stress conditions, the accumulation of reactive oxygen species (ROS) was remarkably decreased in the transgenic lines as compared with the wild type. Moreover, the transcript levels of three stress-responsive genes were increased in the transgenic tobacco lines. Taken together, our results suggest that PtADC plays a key role in drought tolerance, which is, at least partially, attributed to its role in ROS detoxification.

  15. Zymographic detection of cinnamic acid decarboxylase activity.

    PubMed

    Prim, Núria; Pastor, F I Javier; Diaz, Pilar

    2002-11-01

    The manuscript includes a concise description of a new, fast and simple method for detection of cinnamic acid decarboxylase activity. The method is based on a color shift caused a by pH change and may be an excellent procedure for large screenings of samples from natural sources, as it involves no complex sample processing or purification. The method developed can be used in preliminary approaches to biotransformation processes involving detection of hydroxycinnamic acid decarboxylase activity.

  16. AdcAII of Streptococcus pneumoniae Affects Pneumococcal Invasiveness

    PubMed Central

    Brown, Lindsey R.; Gunnell, Steven M.; Cassella, Adam N.; Keller, Lance E.; Scherkenbach, Lisa A.; Mann, Beth; Brown, Matthew W.; Hill, Rebecca; Fitzkee, Nicholas C.; Rosch, Jason W.; Tuomanen, Elaine I.; Thornton, Justin A.

    2016-01-01

    Across bacterial species, metal binding proteins can serve functions in pathogenesis in addition to regulating metal homeostasis. We have compared and contrasted the activities of zinc (Zn2+)-binding lipoproteins AdcA and AdcAII in the Streptococcus pneumoniae TIGR4 background. Exposure to Zn2+-limiting conditions resulted in delayed growth in a strain lacking AdcAII (ΔAdcAII) when compared to wild type bacteria or a mutant lacking AdcA (ΔAdcA). AdcAII failed to interact with the extracellular matrix protein laminin despite homology to laminin-binding proteins of related streptococci. Deletion of AdcA or AdcAII led to significantly increased invasion of A549 human lung epithelial cells and a trend toward increased invasion in vivo. Loss of AdcAII, but not AdcA, was shown to negatively impact early colonization of the nasopharynx. Our findings suggest that expression of AdcAII affects invasiveness of S. pneumoniae in response to available Zn2+ concentrations. PMID:26752283

  17. Condensation of anhydrides or dicarboxylic acids with compounds containing active methylene groups. Part 1: Condensation of phthalic anhydride with acetoacetic and malonic ester

    NASA Technical Reports Server (NTRS)

    Oshkaya, V. P.; Vanag, G. Y.

    1985-01-01

    Phthalic anhydride was condensed with acetoacetic ester in acetic anhydride and triethylamine solution, and when phthalyl chloride was reacted with sodium acetoacetic ester compounds were formed of the phthalide and indandione series: phthalylacetoacetic ester and a derivative of indan-1,3-dione which after boiling with hydrochloric acid yielded indan-1,3-dione. Phthalylmalonic ester was obtained from phthalic anhydride and malonic ester in the presence of triethylamine.

  18. FcWRKY70, a WRKY protein of Fortunella crassifolia, functions in drought tolerance and modulates putrescine synthesis by regulating arginine decarboxylase gene.

    PubMed

    Gong, Xiaoqing; Zhang, Jingyan; Hu, Jianbing; Wang, Wei; Wu, Hao; Zhang, Qinghua; Liu, Ji-Hong

    2015-11-01

    WRKY comprises a large family of transcription factors in plants, but most WRKY members are still poorly understood. In this study, we report functional characterization of a Group III WRKY gene (FcWRKY70) from Fortunella crassifolia. FcWRKY70 was greatly induced by drought and abscisic acid, but slightly or negligibly by salt and cold. Overexpression of FcWRKY70 in tobacco (Nicotiana nudicaulis) and lemon (Citrus lemon) conferred enhanced tolerance to dehydration and drought stresses. Transgenic tobacco and lemon exhibited higher expression levels of ADC (arginine decarboxylase), and accumulated larger amount of putrescine in comparison with wild type (WT). Treatment with D-arginine, an inhibitor of ADC, caused transgenic tobacco plants more sensitive to dehydration. Knock-down of FcWRKY70 in kumquat down-regulated ADC abundance and decreased putrescine level, accompanied by compromised dehydration tolerance. The promoter region of FcADC contained two W-box elements, which were shown to be interacted with FcWRKY70. Taken together, our data demonstrated that FcWRKY70 functions in drought tolerance by, at least partly, promoting production of putrescine via regulating ADC expression.

  19. Conformational analysis, tautomerization, IR, Raman, and NMR studies of benzyl acetoacetate

    NASA Astrophysics Data System (ADS)

    Tayyari, Sayyed Faramarz; Naghavi, Farnaz; Pojhan, Sahar; McClurg, Ryan W.; Sammelson, Robert E.

    2011-02-01

    A complete conformational analysis of the keto and enol forms of benzyl acetoacetate (BAA), a β-dicarbonyl compound, was carried out by ab initio calculations, at the density functional theory (DFT) level. By inspection of all possible conformers and tautomers, 22 stable cis-enol, 28 stable trans-enol, and five keto conformers were obtained. Among all stable cis-enol forms only six of them are engaged in intramolecular hydrogen bond. The hydrogen bond strength of the most stable conformer of BAA is compared with that of acetylacetone (AA) and dimethyl oxaloacetate (DMOA). Harmonic vibrational frequencies of the most stable enol and keto forms and their deuterated analogues were also calculated and compared with the experimental data. According to the theoretical calculations, the hydrogen bond strength of the most stable enol conformer of BAA is 56.7 kJ/mol (calculated at the B3LYP/6-311++G ∗∗ level), about 10 kJ/mol less than that of AA. This weakening of hydrogen bond is consistent with the spectroscopic results. NMR studies indicate that BAA exists mainly as a keto tautomer in all considered solutions. The Gibbs energies for keto/enol tautomerization were calculated at the B3LYP level, with several basis sets, in both gas phase and CH 3CN solution (using PCM model), for the most stable enol and keto conformers.

  20. The stimulation of hepatic gluconeogenesis by acetoacetate precursors. A role for the monocarboxylate translocator.

    PubMed

    Patel, T B; Barron, L L; Olson, M S

    1984-06-25

    The regulation of the gluconeogenic pathway from the 3-carbon precursors pyruvate, lactate, and alanine was investigated in the isolated perfused rat liver. Using pyruvate (less than 1 mM), lactate, or alanine as the gluconeogenic precursor, infusion of the acetoacetate precursors oleate, acetate, or beta-hydroxybutyrate stimulated the rate of glucose production and, in the case of pyruvate (less than 1 mM), the rate of pyruvate decarboxylation. alpha-Cyanocinnamate, an inhibitor of the monocarboxylate transporter, prevented the stimulation of pyruvate decarboxylation and glucose production due to acetate infusion. With lactate as the gluconeogenic precursor, acetate infusion in the presence of L-carnitine stimulated the rate of gluconeogenesis (100%) and ketogenesis (60%) without altering the tissue acetyl-CoA level usually considered a requisite for the stimulation of gluconeogenesis by fatty acids. Hence, our studies suggest that gluconeogenesis from pyruvate or other substrates which are converted to pyruvate prior to glucose synthesis may be limited or controlled by the rate of entry of pyruvate into the mitochondrial compartment on the monocarboxylate translocator.

  1. ADC and TDC implemented using FPGA

    SciTech Connect

    Wu, Jinyuan; Hansen, Sten; Shi, Zonghan; /Fermilab

    2007-11-01

    Several tests of FPGA devices programmed as analog waveform digitizers are discussed. The ADC uses the ramping-comparing scheme. A multi-channel ADC can be implemented with only a few resistors and capacitors as external components. A periodic logic levels are shaped by passive RC network to generate exponential ramps. The FPGA differential input buffers are used as comparators to compare the ramps with the input signals. The times at which these ramps cross the input signals are digitized by time-to-digital-converters (TDCs) implemented within the FPGA. The TDC portion of the logic alone has potentially a broad range of HEP/nuclear science applications. A 96-channel TDC card using FPGAs as TDCs being designed for the Fermilab MIPP electronics upgrade project is discussed. A deserializer circuit based on multisampling circuit used in the TDC, the 'Digital Phase Follower' (DPF) is also documented.

  2. Physiological and biochemical effects of allelochemical ethyl 2-methyl acetoacetate (EMA) on cyanobacterium Microcystis aeruginosa.

    PubMed

    Hong, Yu; Hu, Hong-Ying; Li, Feng-Min

    2008-10-01

    The physiological and biochemical effects of an allelochemical ethyl 2-methyl acetoacetate (EMA) isolated from reed (Phragmites communis) on bloom-forming cyanobacterium, Microcystis aeruginosa, were investigated. EMA significantly inhibited the growth of M. aeruginosa in a concentration-dependent way. The metabolic indices (represented by esterase and total dehydrogenase activities), the cellular redox status (represented by the level of reactive oxygen species (ROS)), and the oxidative damage index (represented by the content of malondialdehyde (MDA), the product of membrane lipid peroxidation) were used to evaluate the physiological and biochemical changes in M. aeruginosa after EMA exposure. Esterase activity in M. aeruginosa did not change (P>0.05) after 2 h of exposure to EMA, but increased greatly after 24 and 48 h (P<0.05). EMA exposure (>0.5 mg L(-1)) resulted in a remarkable loss of total dehydrogenase activity in M. aeruginosa after 4 h (P<0.01), but an increase after 40 h (P<0.05). EMA caused a great increase in ROS level of the algal cells. At high EMA concentration (4 mg L(-1)), the ROS level was remarkably elevated to 1.91 times as much as that in the controls after 2 h. Increases in the ROS level also occurred after 24 and 48 h. The increase in lipid peroxidation of M. aeruginosa was dependent upon EMA concentration and the exposure time. After 40 h of exposure, the MDA content at 4 mg L(-1) of EMA reached approximately 3.5 times (P<0.01) versus the controls. These results suggest that the cellular structure and metabolic activity of M. aeruginosa are influenced by EMA; the increased metabolic activity perhaps reflects the fact that the resistance of cellular response system to the stress from EMA is initiated during EMA exposure, and the oxidative damage induced by EMA via the oxidation of ROS may be an important factor responsible for the inhibition of EMA on the growth of M. aeruginosa.

  3. Novel and efficient ADC concept for BlackGEM telescope

    NASA Astrophysics Data System (ADS)

    ter Horst, Rik; Kragt, Jan; Lesman, Dirk; Navarro, Ramon

    2016-07-01

    Ground based telescopes suffer from Atmospheric Dispersion that can be compensated for with an Atmospheric Dispersion Corrector (ADC). In the BlackGEM array of 650 mm diameter telescopes, the ADC is fully integrated in the three-lens field corrector and requires lateral displacement of only one lens for a full correction of the Atmospheric Dispersion. This concept results in a very compact and efficient ADC design without the need for any additional optical components. This paper describes the optical trade-offs, optical design and optimization, as well as the mechanical design and implementation of this novel ADC solution.

  4. Elevation of arginine decarboxylase-dependent putrescine production enhances aluminum tolerance by decreasing aluminum retention in root cell walls of wheat.

    PubMed

    Yu, Yan; Jin, Chongwei; Sun, Chengliang; Wang, Jinghong; Ye, Yiquan; Lu, Lingli; Lin, Xianyong

    2015-12-15

    Aluminum (Al) stress induces putrescine (Put) accumulation in several plants and this response is proposed to alleviate Al toxicity. However, the mechanisms underlying this alleviation remain largely unknown. Here, we show that exposure to Al clearly increases Put accumulation in the roots of wheat plants (Triticum aestivum L. 'Xi Aimai-1') and that this was accompanied by significant increase in the activity of arginine decarboxylase (ADC), a Put producing enzyme. Application of an ADC inhibitor (d-arginine) terminated the Al-induced Put accumulation, indicating that increased ADC activity may be responsible for the increase in Put accumulation in response to Al. The d-arginine treatment also increased the Al-induced accumulation of cell wall polysaccharides and the degree of pectin demethylation in wheat roots. Thus, it elevated Al retention in cell walls and exacerbated Al accumulation in roots, both of which aggravate Al toxicity in wheat plants. The opposite effects were true for exogenous Put application. These results suggest that ADC-dependent Put accumulation plays important roles in providing protection against Al toxicity in wheat plants through decreasing cell wall polysaccharides and increasing the degree of pectin methylation, thus decreasing Al retention in the cell walls.

  5. Chlorine Dioxide-Iodide-Methyl Acetoacetate Oscillation Reaction Investigated by UV-Vis and Online FTIR Spectrophotometric Method

    PubMed Central

    Shi, Laishun; Wang, Xiaomei; Li, Na; Liu, Jie; Yan, Chunying

    2012-01-01

    In order to study the chemical oscillatory behavior and mechanism of a new chlorine dioxide-iodide ion-methyl acetoacetate reaction system, a series of experiments were done by using UV-Vis and online FTIR spectrophotometric method. The initial concentrations of methyl acetoacetate, chlorine dioxide, potassium iodide, and sulfuric acid and the pH value have great influence on the oscillation observed at wavelength of 289 nm. There is a preoscillatory or induction period, and the amplitude and the number of oscillations are associated with the initial concentration of reactants. The equations for the triiodide ion reaction rate changing with reaction time and the initial concentrations in the oscillation stage were obtained. Oscillation reaction can be accelerated by increasing temperature. The apparent activation energies in terms of the induction period and the oscillation period were 26.02 KJ/mol and 17.65 KJ/mol, respectively. The intermediates were detected by the online FTIR analysis. Based upon the experimental data in this work and in the literature, a plausible reaction mechanism was proposed for the oscillation reaction. PMID:22454614

  6. Chlorine dioxide-iodide-methyl acetoacetate oscillation reaction investigated by UV-vis and online FTIR spectrophotometric method.

    PubMed

    Shi, Laishun; Wang, Xiaomei; Li, Na; Liu, Jie; Yan, Chunying

    2012-01-01

    In order to study the chemical oscillatory behavior and mechanism of a new chlorine dioxide-iodide ion-methyl acetoacetate reaction system, a series of experiments were done by using UV-Vis and online FTIR spectrophotometric method. The initial concentrations of methyl acetoacetate, chlorine dioxide, potassium iodide, and sulfuric acid and the pH value have great influence on the oscillation observed at wavelength of 289 nm. There is a preoscillatory or induction period, and the amplitude and the number of oscillations are associated with the initial concentration of reactants. The equations for the triiodide ion reaction rate changing with reaction time and the initial concentrations in the oscillation stage were obtained. Oscillation reaction can be accelerated by increasing temperature. The apparent activation energies in terms of the induction period and the oscillation period were 26.02 KJ/mol and 17.65 KJ/mol, respectively. The intermediates were detected by the online FTIR analysis. Based upon the experimental data in this work and in the literature, a plausible reaction mechanism was proposed for the oscillation reaction.

  7. ADC evaluation of the corticospinal tract in multiple sclerosis.

    PubMed

    Inal, Mikail; Unal, Birsen; Kala, Ibrahim; Turkel, Yakup; Bilgili, Yasemin Karadeniz

    2015-06-01

    Apparent diffusion coefficient (ADC) values derived from diffusion-weighted MR imaging (DWI) provide important information about tissues. The goal of this study was to evaluate the ADC values in the corticospinal tract regions in multiple sclerosis (MS). The ADC values of 42 patients with multiple sclerosis and 46 healthy people were measured. The ADC values in the corticospinal tract at the capsula interna posterior crus from six points and mesencephalon from three points bilaterally in MS patients were compared with those of controls. An ANOVA post hoc test was used to analyse the differences in mean ADC values between the MS and control groups. The mean ADC values of the right (p = 0.008) and left internal capsules (p = 0.000) and right (p = 0.002) and left mesencephalons (p = 0.044) in MS patients were significantly lower than in the control group. There was no significant difference between the right and left side ADC values in MS (p = 0.313 vs. p = 0.223) and control groups (p = 0.756 vs. p = 0.105), respectively. The mean ADC values of the corticospinal tract in MS patients were significantly lower than in the control group. This decreased diffusion may be the result of cellular infiltration due to inflammation, cytotoxic oedema, demyelination or remyelination processes.

  8. Serial pixel analog-to-digital converter (ADC)

    NASA Astrophysics Data System (ADS)

    Larson, Eric D.

    2010-02-01

    This method reduces the data path from the counter to the pixel register of the analog-to-digital converter (ADC) from as many as 10 bits to a single bit. The reduction in data path width is accomplished by using a coded serial data stream similar to a pseudo random number (PRN) generator. The resulting encoded pixel data is then decoded into a standard hexadecimal format before storage. The high-speed serial pixel ADC concept is based on the single-slope integrating pixel ADC architecture. Previous work has described a massively parallel pixel readout of a similar architecture. The serial ADC connection is similar to the state-of-the art method with the exception that the pixel ADC register is a shift register and the data path is a single bit. A state-of-the-art individual-pixel ADC uses a single-slope charge integration converter architecture with integral registers and "one-hot" counters. This implies that parallel data bits are routed among the counter and the individual on-chip pixel ADC registers. The data path bit-width to the pixel is therefore equivalent to the pixel ADC bit resolution.

  9. Induction of aromatic-L-amino acid decarboxylase by decarboxylase inhibitors in idiopathic parkinsonism.

    PubMed

    Boomsma, F; Meerwaldt, J D; Man in 't Veld, A J; Hovestadt, A; Schalekamp, M A

    1989-06-01

    We evaluated the effect of administration of L-dopa, alone or in combination with a peripheral decarboxylase inhibitor, on plasma levels of aromatic-L-amino acid decarboxylase (ALAAD). After single-dose administration of L-dopa plus benserazide (Madopar) in healthy subjects and in chronically treated patients with parkinsonism, plasma ALAAD followed for 2 to 3 hours fell, but returned to predosing levels within 90 minutes. Four groups of patients with idiopathic parkinsonism were studied during chronic treatment: Group I, no L-dopa treatment (n = 31); Group II, L-dopa alone (n = 15); Group III, L-dopa plus benserazide (n = 28); and Group IV, L-dopa plus carbidopa (Sinemet, n = 30). Plasma ALAAD 2 hours after dosing was normal in Groups I and II. ALAAD was increased threefold in Groups III and IV, suggesting induction of ALAAD by the coadministration of a peripheral decarboxylase inhibitor. In a study of 3 patients in whom L-dopa/benserazide was started, plasma ALAAD rose gradually over 3 to 4 weeks. Further detailed pharmacokinetic studies of L-dopa, dopamine, and ALAAD in plasma and cerebrospinal fluid are required to determine if the apparent ALAAD induction by a peripheral decarboxylase inhibitor may be related to the loss of clinical efficacy of combination therapy in some patients and how it is related to end-of-dose deterioration and on-off phenomena.

  10. High performance 14-bit pipelined redundant signed digit ADC

    NASA Astrophysics Data System (ADS)

    Narula, Swina; Pandey, Sujata

    2016-03-01

    A novel architecture of a pipelined redundant-signed-digit analog to digital converter (RSD-ADC) is presented featuring a high signal to noise ratio (SNR), spurious free dynamic range (SFDR) and signal to noise plus distortion (SNDR) with efficient background correction logic. The proposed ADC architecture shows high accuracy with a high speed circuit and efficient utilization of the hardware. This paper demonstrates the functionality of the digital correction logic of 14-bit pipelined ADC at each 1.5 bit/stage. This prototype of ADC architecture accounts for capacitor mismatch, comparator offset and finite Op-Amp gain error in the MDAC (residue amplification circuit) stages. With the proposed architecture of ADC, SNDR obtained is 85.89 dB, SNR is 85.9 dB and SFDR obtained is 102.8 dB at the sample rate of 100 MHz. This novel architecture of digital correction logic is transparent to the overall system, which is demonstrated by using 14-bit pipelined ADC. After a latency of 14 clocks, digital output will be available at every clock pulse. To describe the circuit behavior of the ADC, VHDL and MATLAB programs are used. The proposed architecture is also capable of reducing the digital hardware. Silicon area is also the complexity of the design.

  11. Spermine facilitates recovery from drought but does not confer drought tolerance in transgenic rice plants expressing Datura stramonium S-adenosylmethionine decarboxylase.

    PubMed

    Peremarti, Ariadna; Bassie, Ludovic; Christou, Paul; Capell, Teresa

    2009-06-01

    Polyamines are known to play important roles in plant stress tolerance but it has been difficult to determine precise functions for each type of polyamine and their interrelationships. To dissect the roles of putrescine from the higher polyamines spermidine and spermine, we generated transgenic rice plants constitutively expressing a heterologous S-adenosylmethionine decarboxylase (SAMDC) gene from Datura stramonium so that spermidine and spermine levels could be investigated while maintaining a constant putrescine pool. Whereas transgenic plants expressing arginine decarboxylase (ADC) produced higher levels of putrescine, spermidine and spermine, and were protected from drought stress, transgenic plants expressing SAMDC produced normal levels of putrescine and showed drought symptoms typical of wild type plants under stress, but the transgenic plants showed a much more robust recovery on return to normal conditions (90% full recovery compared to 25% partial recovery for wild type plants). At the molecular level, both wild type and transgenic plants showed transient reductions in the levels of endogenous ADC1 and SAMDC mRNA, but only wild type plants showed a spike in putrescine levels under stress. In transgenic plants, there was no spike in putrescine but a smooth increase in spermine levels at the expense of spermidine. These results confirm and extend the threshold model for polyamine activity in drought stress, and attribute individual roles to putrescine, spermidine and spermine.

  12. Metal-free arylation of ethyl acetoacetate with hypervalent diaryliodonium salts: an immediate access to diverse 3-aryl-4(1H)-quinolones.

    PubMed

    Monastyrskyi, Andrii; Namelikonda, Niranjan K; Manetsch, Roman

    2015-03-06

    A clean arylation protocol of ethyl acetoacetate was developed using hypervalent diaryliodonium salts under mild and metal-free conditions. The scope of the reaction, using symmetric and unsymmetric iodonium salts with varying sterics and electronics, was examined. Further, this method has been applied for the synthesis of antimalarial compound ELQ-300, which is currently in preclinical development.

  13. Role of glutamate decarboxylase-like protein 1 (GADL1) in taurine biosynthesis.

    PubMed

    Liu, Pingyang; Ge, Xiaomei; Ding, Haizhen; Jiang, Honglin; Christensen, Bruce M; Li, Jianyong

    2012-11-30

    This manuscript concerns the tissue-specific transcription of mouse and cattle glutamate decarboxylase-like protein 1 (GADL1) and the biochemical activities of human GADL1 recombinant protein. Bioinformatic analysis suggested that GADL1 appears late in evolution, only being found in reptiles, birds, and mammals. RT-PCR determined that GADL1 mRNA is transcribed at high levels in mouse and cattle skeletal muscles and also in mouse kidneys. Substrate screening determined that GADL1, unlike its name implies, has no detectable GAD activity, but it is able to efficiently catalyze decarboxylation of aspartate, cysteine sulfinic acid, and cysteic acid to β-alanine, hypotaurine, and taurine, respectively. Western blot analysis verified the presence of GADL1 in mouse muscles, kidneys, C2C12 myoblasts, and C2C12 myotubes. Incubation of the supernatant of fresh muscle or kidney extracts with cysteine sulfinic acid resulted in the detection of hypotaurine or taurine in the reaction mixtures, suggesting the possible involvement of GADL1 in taurine biosynthesis. However, when the tissue samples were incubated with aspartate, no β-alanine production was observed. We proposed several possibilities that might explain the inactivation of ADC activity of GADL1 in tissue protein extracts. Although β-alanine-producing activity was not detected in the supernatant of tissue protein extracts, its potential role in β-alanine synthesis cannot be excluded. There are several inhibitors of the ADC activity of GADL1 identified. The discovery of GADL1 biochemical activities, in conjunction with its expression and activities in muscles and kidneys, provides some tangible insight toward establishing its physiological function(s).

  14. Identification of the putrescine biosynthetic genes in Pseudomonas aeruginosa and characterization of agmatine deiminase and N-carbamoylputrescine amidohydrolase of the arginine decarboxylase pathway.

    PubMed

    Nakada, Yuji; Itoh, Yoshifumi

    2003-03-01

    Putrescine can be synthesized either directly from ornithine by ornithine decarboxylase (ODC; the speC product) or indirectly from arginine via arginine decarboxylase (ADC; the speA product). The authors identified the speA and speC genes in Pseudomonas aeruginosa PAO1. The activities of the two decarboxylases were similar and each enzyme alone appeared to direct sufficient formation of the polyamine for normal growth. A mutant defective in both speA and speC was a putrescine auxotroph. In this strain, agmatine deiminase (the aguA product) and N-carbamoylputrescine amidohydrolase (the aguB product), which were initially identified as the catabolic enzymes of agmatine, biosynthetically convert agmatine to putrescine in the ADC pathway: a double mutant of aguAB and speC was a putrescine auxotroph. AguA was purified as a homodimer of 43 kDa subunits and AguB as a homohexamer of 33 kDa subunits. AguA specifically deiminated agmatine with K(m) and K(cat) values of 0.6 mM and 4.2 s(-1), respectively. AguB was specific to N-carbamoylputrescine and the K(m) and K(cat) values of the enzyme for the substrate were 0.5 mM and 3.3 s(-1), respectively. Whereas AguA has no structural relationship to any known C-N hydrolases, AguB is a protein of the nitrilase family that performs thiol-assisted catalysis. Inhibition by SH reagents and the conserved cysteine residue in AguA and its homologues suggested that this enzyme is also involved in thiol-mediated catalysis.

  15. On-chip ADC for infrared focal plane arrays

    NASA Astrophysics Data System (ADS)

    Gao, Lei; Chen, Guo-qiang; Wang, Pan; Ding, Rui-jun

    2013-09-01

    This paper presents a low power and small area analog-digital converter (ADC) for infrared focal plane arrays (IRFPA) readout integrated circuit (ROIC). Successive approximation register (SAR) ADC architecture is used in this IRFPA readout integrated circuit. Each column of the IRFPA shares one SAR ADC. The most important part is the three-level DAC. Compared to the previous design, this three-level DAC needs smaller area, has lower power, and more suitable for IRFPA ROIC. In this DAC, its most significant bit (MSB) sub-DAC uses charge scaling, while the least significant bit (LSB) sub-DAC uses voltage scaling. Where the MSB sub-DAC consists of a four-bit charge scaling DAC and a five-bit sub-charge scaling DAC. We need to put a scaling capacitor Cs between these two sub-DACs. Because of the small area, we have more design methods to make the ADC has a symmetrical structure and has higher accuracy. The ADC also needs a high resolution comparator. In this design the comparator uses three-stage operational amplifier structure to have a 77dB differential gain. As the IR focal plane readout circuit signal is stepped DC signal, the circuit design time without adding the sample and hold circuit, so we can use a DC signal instead of infrared focal plane readout circuit output analog signals to be simulated. The simulation result shows that the resolution of the ADC is 12 bit.

  16. Dopa decarboxylase activity of the living human brain

    SciTech Connect

    Gjedde, A.; Reith, J.; Dyve, S.; Leger, G.; Guttman, M.; Diksic, M.; Evans, A.; Kuwabara, H. )

    1991-04-01

    Monoaminergic neurons use dopa decarboxylase to form dopamine from L-3,4-dihydroxyphenylalanine (L-dopa). We measured regional dopa decarboxylase activity in brains of six healthy volunteers with 6-({sup 18}F)fluoro-L-dopa and positron emission tomography. We calculated the enzyme activity, relative to its Km, with a kinetic model that yielded the relative rate of conversion of 6-({sup 18}F)fluoro-L-dopa to ({sup 18}F)fluorodopamine. Regional values of relative dopa decarboxylase activity ranged from nil in occipital cortex to 1.9 h-1 in caudate nucleus and putamen, in agreement with values obtained in vitro.

  17. Three Distinct Glutamate Decarboxylase Genes in Vertebrates

    PubMed Central

    Grone, Brian P.; Maruska, Karen P.

    2016-01-01

    Gamma-aminobutyric acid (GABA) is a widely conserved signaling molecule that in animals has been adapted as a neurotransmitter. GABA is synthesized from the amino acid glutamate by the action of glutamate decarboxylases (GADs). Two vertebrate genes, GAD1 and GAD2, encode distinct GAD proteins: GAD67 and GAD65, respectively. We have identified a third vertebrate GAD gene, GAD3. This gene is conserved in fishes as well as tetrapods. We analyzed protein sequence, gene structure, synteny, and phylogenetics to identify GAD3 as a homolog of GAD1 and GAD2. Interestingly, we found that GAD3 was lost in the hominid lineage. Because of the importance of GABA as a neurotransmitter, GAD3 may play important roles in vertebrate nervous systems. PMID:27461130

  18. Keto-isovalerate decarboxylase enzymes and methods of use thereof

    DOEpatents

    McElvain, Jessica; O'Keefe, Daniel P.; Paul, Brian James; Payne, Mark S.; Rothman, Steven Cary; He, Hongxian

    2016-01-19

    Provided herein are polypeptides and polynucleotides encoding such polypeptides which have ketoisovalerate decarboxylase activity. Also provided are recombinant host cells comprising such polypeptides and polynucleotides and methods of use thereof.

  19. Exact analytical results for ADC with oscillating diffusion sensitizing gradients

    PubMed Central

    Sukstanskii, A.L.

    2013-01-01

    The apparent diffusion coefficient (ADC) is analyzed for the case of oscillating diffusion sensitizing gradients. Exact analytical expressions are obtained in the high-frequency expansion of the ADC for an arbitrary number of oscillations N. These expressions are universal and valid for arbitrary system geometry. The validity conditions of the high-frequency expansion of ADC are obtained in the framework of a simple 1D model of restricted diffusion. These conditions are shown to be substantially different for cos- and sin-type gradients: for the cos-type gradients, the high-frequency expansion is valid when the period of a single oscillation is smaller than the characteristic diffusion time, the frequency dependence of ADC being practically the same for any N. In contrast, for the sin-type gradients, the high-frequency regime can be achieved only when the total diffusion time is smaller than the characteristic diffusion time, the frequency dependence of ADC being different for different N. PMID:23876779

  20. Effects of a novel allelochemical ethyl 2-methyl acetoacetate (EMA) on the ultrastructure and pigment composition of cyanobacterium Microcystis aeruginosa.

    PubMed

    Hong, Yu; Huang, Jing-Jing; Hu, Hong-Ying

    2009-10-01

    Allelochemical ethyl 2-methyl acetoacetate (EMA) can significantly inhibit the growth of bloom-forming Microcystis aeruginosa. In order to assess the implication of the damage of EMA on the algal photosynthetic apparatus, the effects of EMA on the algal ultrastructure and pigment composition were investigated. At initial exposure time to EMA (0-40 h), algal allophycocyanin, phycoerythrin and carotenoid degraded firstly; chlorophyll a increased, especially by 47% in the algae exposed to 2 mg L(-1) of EMA; phycocyanin was not significantly affected; lipid bodies increased remarkably. After 40 h of EMA exposure, chlorophyll a decreased gradually, especially by 45% in the algae exposed to 4 mg L(-1) of EMA; lipid bodies greatly reduced but cyanophycin granules accumulated; thylakoid structures were dissolved or disappeared with the presence of numerous vacuoles. These results showed that all ophycocyanin, phycoerythrin and carotenoid were more sensitive to EMA than other pigments, the cells of M. aeruginosa was stressed by EMA with the occurrence of cyanophycin granules and the photosynthesis pigments and ultrastructure of M. aeruginosa were quickly destroyed by EMA with exposure time increasing.

  1. Digital background calibration of charge pump based pipelined ADC

    NASA Astrophysics Data System (ADS)

    Singh, Anil; Agarwal, Alpana

    2016-11-01

    In the presented work, digital background calibration of a charge pump based pipelined ADC is presented. A 10-bit 100 MS/s pipelined ADC is designed using TSMC 0.18 µm CMOS technology operating on a 1.8 V power supply voltage. A power efficient opamp-less charge pump based technique is chosen to achieve the desired stage voltage gain of 2 and digital background calibration is used to calibrate the inter-stage gain error. After calibration, the ADC achieves an SNDR of 66.78 dB and SFDR of 79.3 dB. Also, DNL improves to +0.6/-0.4 LSB and INL improves from +9.3/-9.6 LSB to within ±0.5 LSB, consuming 16.53 mW of power.

  2. Mapping of glutamic acid decarboxylase (GAD) genes

    SciTech Connect

    Edelhoff, S.; Adler, D.A.; Disteche, C.M.; Grubin, C.E.; Karlsen, A.E.; Lernmark, A.; Foster, D. )

    1993-07-01

    Glutamic acid decarboxylase (GAD) catalyzes the synthesis of [gamma]-aminobutyric acid (GABA), which is known as a major inhibitory neurotransmitter in the central nervous system (CNS), but is also present outside the CNS. Recent studies showed that GAD is the major target of autoantibodies associated with the development of insulin-dependent diabetes mellitus and of the rare stiff man syndrome. Studies of GAD expression have demonstrated multiple transcripts, suggesting several isoforms of GAD. In this study, three different genes were mapped by in situ hybridization to both human and mouse chromosomes. The GAD1 gene was mapped to human chromosome 2q31 and to mouse chromosome 2D in a known region of conservation between human and mouse. GAD2, previously mapped to human chromosome 10p11.2-p12, was mapped to mouse chromosome 2A2-B, which identifies a new region of conservation between human and mouse chromosomes. A potential GAD3 transcript was mapped to human chromosome 22q13 and to mouse chromosome 15E in a known region of conservation between human and mouse. It is concluded that the GAD genes may form a family with as many as three related members. 30 refs., 5 figs.

  3. Post-transcriptional regulation of ornithine decarboxylase

    PubMed Central

    Nowotarski, Shannon L.; Origanti, Sofia; Shantz, Lisa M.

    2013-01-01

    Activity of the polyamine biosynthetic enzyme ornithine decarboxylase (ODC), and intracellular levels of ODC protein are controlled very tightly. Numerous studies have described ODC regulation at the levels of transcription, translation and protein degradation in normal cells, and dysregulation of these processes in response to oncogenic stimuli. Although post-transcriptional regulation of ODC has been well-documented, the RNA binding proteins (RBPs) that interact with ODC mRNA and control synthesis of the ODC protein have not been defined. Using Ras-transformed rat intestinal epithelial cells (Ras12V cells) as a model, we have begun identifying the RBPs that associate with the ODC transcript. Binding of RBPs could potentially regulate ODC synthesis by either changing mRNA stability or rate of mRNA translation. Techniques for measuring RBP binding and translation initiation are described here. Targeting control of ODC translation or mRNA decay could be a valuable method of limiting polyamine accumulation and subsequent tumor development in a variety of cancers. PMID:21318880

  4. A Porphodimethene Chemical Inhibitor of Uroporphyrinogen Decarboxylase

    PubMed Central

    Yip, Kenneth W.; Zhang, Zhan; Sakemura-Nakatsugawa, Noriko; Huang, Jui-Wen; Vu, Nhu Mai; Chiang, Yi-Kun; Lin, Chih-Lung; Kwan, Jennifer Y. Y.; Yue, Shijun; Jitkova, Yulia; To, Terence; Zahedi, Payam; Pai, Emil F.; Schimmer, Aaron D.; Lovell, Jonathan F.; Sessler, Jonathan L.; Liu, Fei-Fei

    2014-01-01

    Uroporphyrinogen decarboxylase (UROD) catalyzes the conversion of uroporphyrinogen to coproporphyrinogen during heme biosynthesis. This enzyme was recently identified as a potential anticancer target; its inhibition leads to an increase in reactive oxygen species, likely mediated by the Fenton reaction, thereby decreasing cancer cell viability and working in cooperation with radiation and/or cisplatin. Because there is no known chemical UROD inhibitor suitable for use in translational studies, we aimed to design, synthesize, and characterize such a compound. Initial in silico-based design and docking analyses identified a potential porphyrin analogue that was subsequently synthesized. This species, a porphodimethene (named PI-16), was found to inhibit UROD in an enzymatic assay (IC50 = 9.9 µM), but did not affect porphobilinogen deaminase (at 62.5 µM), thereby exhibiting specificity. In cellular assays, PI-16 reduced the viability of FaDu and ME-180 cancer cells with half maximal effective concentrations of 22.7 µM and 26.9 µM, respectively, and only minimally affected normal oral epithelial (NOE) cells. PI-16 also combined effectively with radiation and cisplatin, with potent synergy being observed in the case of cisplatin in FaDu cells (Chou-Talalay combination index <1). This work presents the first known synthetic UROD inhibitor, and sets the foundation for the design, synthesis, and characterization of higher affinity and more effective UROD inhibitors. PMID:24587102

  5. Properties of oxaloacetate decarboxylase from Veillonella parvula.

    PubMed Central

    Ng, S K; Wong, M; Hamilton, I R

    1982-01-01

    Oxaloacetate decarboxylase was purified to 136-fold from the oral anaerobe Veillonella parvula. The purified enzyme was substantially free of contaminating enzymes or proteins. Maximum activity of the enzyme was exhibited at pH 7.0 for both carboxylation and decarboxylation. At this pH, the Km values for oxaloacetate and Mg2+ were at 0.06 and 0.17 mM, respectively, whereas the Km values for pyruvate, CO2, and Mg2+ were 3.3, 1.74, and 1.85 mM, respectively. Hyperbolic kinetics were observed with all of the aforementioned compounds. The Keq' was 2.13 X 10(-3) mM-1 favoring the decarboxylation of oxaloacetate. In the carboxylation step, avidin, acetyl coenzyme A, biotin, and coenzyme A were not required. ADP and NADH had no effect on either the carboxylation or decarboxylation step, but ATP inhibited the carboxylation step competitively and the decarboxylation step noncompetitively. These types of inhibition fitted well with the overall lactate metabolism of the non-carbohydrate-fermenting anaerobe. PMID:7076619

  6. Methods to Design and Synthesize Antibody-Drug Conjugates (ADCs)

    PubMed Central

    Yao, Houzong; Jiang, Feng; Lu, Aiping; Zhang, Ge

    2016-01-01

    Antibody-drug conjugates (ADCs) have become a promising targeted therapy strategy that combines the specificity, favorable pharmacokinetics and biodistributions of antibodies with the destructive potential of highly potent drugs. One of the biggest challenges in the development of ADCs is the application of suitable linkers for conjugating drugs to antibodies. Recently, the design and synthesis of linkers are making great progress. In this review, we present the methods that are currently used to synthesize antibody-drug conjugates by using thiols, amines, alcohols, aldehydes and azides. PMID:26848651

  7. Parallel data analysis in a multichannel flash-ADC-system

    SciTech Connect

    Eckerlin, G.; Elsen, E.; Schmitt, H.V.D.; Wagner, A.; Walter, P.V.; Zimmer, M.

    1987-02-01

    Parallel analysis of drift chamber signals with M68000 processors has proven to be an efficient way to deal with the tremendous data flow generated by high speed (100 MHz) Flash-ADCs in real time. The authors report on the experience gained with a network of 34 processors, placed in 3 VME crates, to read out the 3072 Flash-ADC channels of the JADE Jet-Chamber at PETRA (1). The properties of such a system are compared to more conventional readout schemes for drift chambers.

  8. Characterization of a second lysine decarboxylase isolated from Escherichia coli.

    PubMed Central

    Kikuchi, Y; Kojima, H; Tanaka, T; Takatsuka, Y; Kamio, Y

    1997-01-01

    We report here on the existence of a new gene for lysine decarboxylase in Escherichia coli K-12. The hybridization experiments with a cadA probe at low stringency showed that the homologous region of cadA was located in lambda Kohara phage clone 6F5 at 4.7 min on the E. coli chromosome. We cloned the 5.0-kb HindIII fragment of this phage clone and sequenced the homologous region of cadA. This region contained a 2,139-nucleotide open reading frame encoding a 713-amino-acid protein with a calculated molecular weight of 80,589. Overexpression of the protein and determination of its N-terminal amino acid sequence defined the translational start site of this gene. The deduced amino acid sequence showed 69.4% identity to that of lysine decarboxylase encoded by cadA at 93.7 min on the E. coli chromosome. In addition, the level of lysine decarboxylase activity increased in strains carrying multiple copies of the gene. Therefore, the gene encoding this lysine decarboxylase was designated Idc. Analysis of the lysine decarboxylase activity of strains containing cadA, ldc, or cadA ldc mutations indicated that ldc was weakly expressed under various conditions but is a functional gene in E. coli. PMID:9226257

  9. High Resolution Transferred Substrate HBT Microwave/RF ADCs

    DTIC Science & Technology

    2007-11-02

    of wideband delta sigma ADCs using high speed Indium Phosphide bipolar transistors . 15. SUBJECTTERMS DISTRIBUTION STATEMENTA Approved for Public...kept below several hundred transistors , at high level only a single-bit internal quantizer is feasible. Secondly, although the transferred-substrate... transistor counts. Instead, in this program, higher resolutions were sought through the highest possible clock frequencies. Transferred-substrate HBTs

  10. ADC's Insertion Devices and Magnetic Measurement Systems Capabilities

    NASA Astrophysics Data System (ADS)

    Deyhim, A.; Kulesza, J.

    2013-03-01

    In this paper Advance Design Consulting USA, Inc. (ADC) will discuss ADC's major improved capabilities for building Wiggler Insertion Devices, Undulator Planar Devices, Elliptical Polarizing Undulators (EPU), In-Vacuum Undulators (IVU), Cryogenically Cooled in-vacuum Undulators (CPMU), Super Conductive Undulator, and Insertion Device Magnetic Measurement Systems. ADC has designed, built and delivered Insertion Devices and Magnetic Measurement Systems to such facilities as MAX-lab (two EPUs, a Planar, and Measurement System), ALBA and ASP (Wigglers), BNL (CPMU), SSRF (two IVUs and a Measurement System), PAL (one IVU and Measurement System), NSRRC (one 4m EPU), and SRC (Planar and EPU). ADC's magnetic field measurement system is a sophisticated and sensitive machine for the measurement of magnetic fields in undulators (Planar and EPU), wigglers and in-vacuum ID units. The magnetic fields are measured using 3 axis hall-effect probes, mounted orthogonally, to a thin wand. The wand is mounted to a carriage that rides on vacuum air bearings. The base is granite. A flip coil is provided on two vertical towers with X, Y and Theta axes. Special software is provided to assist in homing, movement, and data collection and analysis.

  11. Pseudo Asynchronous Level Crossing adc for ecg Signal Acquisition.

    PubMed

    Marisa, T; Niederhauser, T; Haeberlin, A; Wildhaber, R A; Vogel, R; Goette, J; Jacomet, M

    2017-02-07

    A new pseudo asynchronous level crossing analogue-to-digital converter (adc) architecture targeted for low-power, implantable, long-term biomedical sensing applications is presented. In contrast to most of the existing asynchronous level crossing adc designs, the proposed design has no digital-to-analogue converter (dac) and no continuous time comparators. Instead, the proposed architecture uses an analogue memory cell and dynamic comparators. The architecture retains the signal activity dependent sampling operation by generating events only when the input signal is changing. The architecture offers the advantages of smaller chip area, energy saving and fewer analogue system components. Beside lower energy consumption the use of dynamic comparators results in a more robust performance in noise conditions. Moreover, dynamic comparators make interfacing the asynchronous level crossing system to synchronous processing blocks simpler. The proposed adc was implemented in [Formula: see text] complementary metal-oxide-semiconductor (cmos) technology, the hardware occupies a chip area of 0.0372 mm(2) and operates from a supply voltage of [Formula: see text] to [Formula: see text]. The adc's power consumption is as low as 0.6 μW with signal bandwidth from [Formula: see text] to [Formula: see text] and achieves an equivalent number of bits (enob) of up to 8 bits.

  12. Spectrofluorimetric protocol for ceftriaxone in commercial formulation and human plasma after condensation with formaldehyde and ethyl acetoacetate.

    PubMed

    Shah, Jasmin; Jan, M Rasul; Shah, Sultan; Naeem, M

    2011-11-01

    A new spectrofluorimetric method has been developed and validated for the quantification of ceftriaxone in bulk powder, pharmaceutical formulations and spiked human plasma. The developed method is reproducible, accurate, sensitive and cost effective. In this method, ceftriaxone was converted into a fluorescent compound by reacting with 0.8 M ethyl acetoacetate and 25% formaldehyde in a buffered medium (pH = 4.2) at 90 °C. The excitation and emission wavelengths of the fluorescent reaction product are 316 nm and 388 nm respectively. Optimization of the experimental conditions affecting the condensation reaction were carefully carried out and the optimum experimental conditions were incorporated in the procedure. The developed method has a broad linear range (0.2-20 μg mL(-1)) with a correlation coefficient of 0.9992. The limit of detection (LOD) and limit of quantification (LOQ) was found to be 1.94 × 10(-2) μg mL(-1) and 6.47 × 10(-2) μg mL(-1) respectively. The common excipients and co-administered drugs were investigated for their interferences effect in the assay. The developed method was validated statistically through recovery studies and successfully applied to ceftriaxone determination in bulk powder, pharmaceutical formulations and spiked human plasma samples. The percent recoveries were found to be in the range of 99.04-100.26% for bulk powder, 98.88-99.92% for pharmaceutical formulations and 94.22-98.48% for spiked human plasma. The results were verified by comparing with reference literature HPLC method and were found in good agreement.

  13. Nucleotide sequence of the pyruvate decarboxylase gene from Zymomonas mobilis.

    PubMed

    Neale, A D; Scopes, R K; Wettenhall, R E; Hoogenraad, N J

    1987-02-25

    Pyruvate decarboxylase (EC 4.1.1.1), the penultimate enzyme in the alcoholic fermentation pathway of Zymomonas mobilis, converts pyruvate to acetaldehyde and carbon dioxide. The complete nucleotide sequence of the structural gene encoding pyruvate decarboxylase from Zymomonas mobilis has been determined. The coding region is 1704 nucleotides long and encodes a polypeptide of 567 amino acids with a calculated subunit mass of 60,790 daltons. The amino acid sequence was confirmed by comparison with the amino acid sequence of a selection of tryptic fragments of the enzyme. The amino acid composition obtained from the nucleotide sequence is in good agreement with that obtained experimentally.

  14. Comparison between activation of ornithine decarboxylase and histidine decarboxylase in rat stomach.

    PubMed

    Ding, X Q; Chen, D; Rosengren, E; Persson, L; Hakanson, R

    1996-03-01

    We compared the responses of rat stomach ornithine decarboxylase (ODC) and histidine decarboxylase (HDC) to food intake, oral treatment with antisecretagogues, NaHCO3, and hypertonic NaCl, antrectomy, intravenous infusion of gastrin-17, the selective cholecystokinin (CCK)-B/gastrin receptor antagonist L-365,260, and the somatostatin analogue RC-160. The serum gastrin concentration and oxyntic mucosal ODC and HDC activities were higher in freely fed rats than in fasted rats. Food intake in fasted rats raised the serum gastrin concentration and the ODC and HDC activities. Ranitidine, omeprazole, and NaHCO3 raised the serum gastrin concentration and activated ODC and HDC. Hypertonic NaCl raised the ODC activity 200-fold, whereas circulating gastrin and HDC activity were increased only moderately. Infusion of gastrin-17 activated HDC but not ODC. L-365,260 prevented the activation of HDC but not of ODC in response to food intake and treatment with omeprazole, NaHCO3, or hypertonic NaCl. Antrectomy prevented the food- and omeprazole-evoked rise in oxyntic mucosal HDC activity but not the rise in ODC activity. RC-160 suppressed HDC activity after food intake and treatment with omeprazole, NaHCO3, or NaCl. In contrast, RC-160 suppressed omeprazole- and NaHCO3-evoked ODC activation but not that evoked by food intake or NaCl. The results support the view that HDC in the oxyntic mucosa is activated by gastrin and suppressed by somatostatin. The induction of ODC is not mediated by gastrin; ODC activation appears to be related to acid inhibition per se or to mucosal maintenance and repair; somatostatin, or rather the lack of it, might contribute to the induction of ODC after acid blockade. The mechanism behind the activation of rat stomach ODC seems to differ depending on the type of stimulus.

  15. Minimal Power Latch for Single-Slope ADCs

    NASA Technical Reports Server (NTRS)

    Hancock, Bruce R.

    2013-01-01

    Column-parallel analog-to-digital converters (ADCs) for imagers involve simultaneous operation of many ADCs. Single-slope ADCs are well adapted to this use because of their simplicity. Each ADC contains a comparator, comparing its input signal level to an increasing reference signal (ramp). When the ramp is equal to the input, the comparator triggers a latch that captures an encoded counter value (code). Knowing the captured code, the ramp value and hence the input signal are determined. In a column-parallel ADC, each column contains only the comparator and the latches; the ramp and code generation are shared. In conventional latch or flip-flop circuits, there is an input stage that tracks the input signal, and this stage consumes switching current every time the input changes. With many columns, many bits, and high code rates, this switching current can be substantial. It will also generate noise that may corrupt the analog signals. A latch was designed that does not track the input, and consumes power only at the instant of latching the data value. The circuit consists of two S-R (set-reset) latches, gated by the comparator. One is set by high data values and the other by low data values. The latches are cross-coupled so that the first one to set blocks the other. In order that the input data not need an inversion, which would consume power, the two latches are made in complementary polarity. This requires complementary gates from the comparator, instead of complementary data values, but the comparator only triggers once per conversion, and usually has complementary outputs to begin with. An efficient CMOS (complementary metal oxide semiconductor) implementation of this circuit is shown in the figure, where C is the comparator output, D is the data (code), and Q0 and Q1 are the outputs indicating the capture of a zero or one value. The latch for Q0 has a negative-true set signal and output, and is implemented using OR-AND-INVERT logic, while the latch for Q1 uses

  16. Base-Promoted Oxidative Cycloaddition Reaction of [60]Fullerene with Ethyl Acetoacetate for C60 Bis-2',3'-dihydrofuran Derivatives: Effect of Bulky Addends.

    PubMed

    Chen, Si; Li, Zong-Jun; Gao, Xiang

    2016-01-04

    The base-promoted oxidative cycloaddition reaction of [60]fullerene with ethyl acetoacetate was investigated. The reaction resulted in C60 bis-2',3'-dihydrofuran derivatives, but only with the trans-1, trans-2, trans-3, and e configurations rather than any cis or trans-4 structures, demonstrating a new regioselectivity resulting from steric influence on C60 bisaddition. In addition, distribution of the bisadducts was complicated by the unsymmetrical nature of the addends, where each individual configuration may consist of several regioisomers.

  17. An ADC for the SAM on the SOAR Telescope

    NASA Astrophysics Data System (ADS)

    Tighe, Roberto; Tokovinin, Andrei; Schurter, Patricio; Martínez, Manuel; Cantarutti, Rolando

    2016-08-01

    SAM (Soar Adaptive-optics Module), the SOAR (Southern Observatory for Astrophysical Research) GLAO facility is in service since 2011, with a UV, 355nm Laser Guide Star (LGS). The atmospheric wavefront error is therefore measured at 355nm and the star images are corrected in the visible range (BVRI bands). An ADC is required for High Resolution imaging at low telescope elevation, especially at shorter wavelengths of the visible spectrum. The ADC is based on 80mm diameter rotating prisms. This compact unit, fully automated, can be inserted or removed from the tightly constrained SAM collimated beam space-envelope, it adjusts to the parallactic angle and corrects the atmospheric dispersion. Here we present the optical and opto-mechanical design, the control design, the operational strategy and performance results obtained from extensive use in on-sky HR Speckle Imaging.

  18. Effect Of Clock Mode On Radiation Hardness Of An ADC

    NASA Technical Reports Server (NTRS)

    Lee, Choon I.; Rax, Bernie G.; Johnston, Allan H.

    1995-01-01

    Report discusses techniques for testing and evaluating effects of total dosages of ionizing radiation on performances of high-resolution successive-approximation analog-to-digital converters (ADCs), without having to test each individual bit or transition. Reduces cost of testing by reducing tests to few critical parametric measurements, from which one determines approximate radiation failure levels providing good approximations of responses of converters for purpose of total-dose-radiation evaluations.

  19. Two-step single slope/SAR ADC with error correction for CMOS image sensor.

    PubMed

    Tang, Fang; Bermak, Amine; Amira, Abbes; Amor Benammar, Mohieddine; He, Debiao; Zhao, Xiaojin

    2014-01-01

    Conventional two-step ADC for CMOS image sensor requires full resolution noise performance in the first stage single slope ADC, leading to high power consumption and large chip area. This paper presents an 11-bit two-step single slope/successive approximation register (SAR) ADC scheme for CMOS image sensor applications. The first stage single slope ADC generates a 3-bit data and 1 redundant bit. The redundant bit is combined with the following 8-bit SAR ADC output code using a proposed error correction algorithm. Instead of requiring full resolution noise performance, the first stage single slope circuit of the proposed ADC can tolerate up to 3.125% quantization noise. With the proposed error correction mechanism, the power consumption and chip area of the single slope ADC are significantly reduced. The prototype ADC is fabricated using 0.18 μ m CMOS technology. The chip area of the proposed ADC is 7 μ m × 500 μ m. The measurement results show that the energy efficiency figure-of-merit (FOM) of the proposed ADC core is only 125 pJ/sample under 1.4 V power supply and the chip area efficiency is 84 k  μ m(2) · cycles/sample.

  20. Development of an ADC radiation tolerance characterization system for the upgrade of the ATLAS LAr calorimeter

    NASA Astrophysics Data System (ADS)

    Liu, Hong-Bin; Chen, Hu-Cheng; Chen, Kai; Kierstead, James; Lanni, Francesco; Takai, Helio; Jin, Ge

    2017-02-01

    ATLAS LAr calorimeter will undergo its Phase-I upgrade during the long shutdown (LS2) in 2018, and a new LAr Trigger Digitizer Board (LTDB) will be designed and installed. Several commercial-off-the-shelf (COTS) multi-channel high-speed ADCs have been selected as possible backups of the radiation tolerant ADC ASICs for the LTDB. To evaluate the radiation tolerance of these backup commercial ADCs, we developed an ADC radiation tolerance characterization system, which includes the ADC boards, data acquisition (DAQ) board, signal generator, external power supplies and a host computer. The ADC board is custom designed for different ADCs, with ADC drivers and clock distribution circuits integrated on board. The Xilinx ZC706 FPGA development board is used as a DAQ board. The data from the ADC are routed to the FPGA through the FMC (FPGA Mezzanine Card) connector, de-serialized and monitored by the FPGA, and then transmitted to the host computer through the Gigabit Ethernet. A software program has been developed with Python, and all the commands are sent to the DAQ board through Gigabit Ethernet by this program. Two ADC boards have been designed for the ADC, ADS52J90 from Texas Instruments and AD9249 from Analog Devices respectively. TID tests for both ADCs have been performed at BNL, and an SEE test for the ADS52J90 has been performed at Massachusetts General Hospital (MGH). Test results have been analyzed and presented. The test results demonstrate that this test system is very versatile, and works well for the radiation tolerance characterization of commercial multi-channel high-speed ADCs for the upgrade of the ATLAS LAr calorimeter. It is applicable to other collider physics experiments where radiation tolerance is required as well. Supported by the U. S. Department of Energy (DE-SC001270)

  1. Blind and reference channel-based time interleaved ADC calibration schemes: a comparison

    NASA Astrophysics Data System (ADS)

    Cimmino, Rosario F.; Centurelli, Francesco; Monsurrò, Pietro; Romano, Francesco; Trifiletti, Alessandro

    2016-07-01

    Many digital background calibration techniques exist which correct for offset, gain, timing and bandwidth mismatches in time-interleaved (TI) ADCs. Some require an additional reference channel, whereas others are blind and rely on the presence of a band where no signal is present (usually around the Nyquist frequency) or exploit other properties of the input signal. Blind calibration techniques, which don't use a reference channel, are suitable for correction of commercial TI-ADCs, or TI-ADC systems using commercial ADCs as channels. Techniques employing additional channels require a more complex layout (especially for the clock tree) and need an additional ADC, whose overhead cost is significant, especially for 2- or 4- channel TI-ADCs. However, we show that the estimation process is faster and more accurate when a reference channel is present, and many different error models can be used (exploiting different points in the accuracy / complexity trade-off).

  2. Recent Advances in Site Specific Conjugations of Antibody Drug Conjugates (ADCs).

    PubMed

    Gao, Wenlong; Zhang, Jingxin; Xiang, Jun; Zhang, Lei; Wu, Chengbin; Dhal, Pradeep K; Chen, Bo

    2016-01-01

    Antibody-drug conjugates (ADCs) take the advantage of antigen specificity of monoclonal antibodies to deliver highly potent cytotoxic drugs selectively to antigen-expressing tumor cells. The recent approval of Adcetris™ and Kadcyla™ as well as emerging data from numerous ongoing clinical trials underscore the role of ADCs as a new therapeutic option for cancer patients. However, conventional conjugation methods generally result in a heterogeneous mixture of ADCs, which can result in significant therapeutic liabilities and can lead to complicated manufacturing processes. The increased understanding from the clinical investigation of current ADCs and site-specific bioconjugation technologies has enabled scientists to accelerate the discovery and development of the next generation ADCs with defined and homogeneous composition. The present manuscript reviews the recent advances and trends in the research and development of novel ADCs obtained by site-specific conjugation method.

  3. A sub-microwatt asynchronous level-crossing ADC for biomedical applications.

    PubMed

    Li, Yongjia; Zhao, Duan; Serdijn, Wouter A

    2013-04-01

    A continuous-time level-crossing analog-to-digital converter (LC-ADC) for biomedical applications is presented. When compared to uniform-sampling (US) ADCs LC-ADCs generate fewer samples for various sparse biomedical signals. Lower power consumption and reduced design complexity with respect to conventional LC-ADCs are achieved due to: 1) replacing the n-bit digital-to-analog converter (DAC) with a 1-bit DAC; 2) splitting the level-crossing detections; and 3) fixing the comparison window. Designed and implemented in 0.18 μm CMOS technology, the proposed ADC uses a chip area of 220 × 203 μm(2). Operating from a supply voltage of 0.8 V, the ADC consumes 313-582 nW from 5 Hz to 5 kHz and achieves an ENOB up to 7.9 bits.

  4. Lotus hairy roots expressing inducible arginine decarboxylase activity.

    PubMed

    Chiesa, María A; Ruiz, Oscar A; Sánchez, Diego H

    2004-05-01

    Biotechnological uses of plant cell-tissue culture usually rely on constitutive transgene expression. However, such expression of transgenes may not always be desirable. In those cases, the use of an inducible promoter could be an alternative approach. To test this hypothesis, we developed two binary vectors harboring a stress-inducible promoter from Arabidopsis thaliana, driving the beta-glucuronidase reporter gene and the oat arginine decarboxylase. Transgenic hairy roots of Lotus corniculatus were obtained with osmotic- and cold-inducible beta-glucuronidase and arginine decarboxylase activities. The increase in the activity of the latter was accompanied by a significant rise in total free polyamines level. Through an organogenesis process, we obtained L. corniculatus transgenic plants avoiding deleterious phenotypes frequently associated with the constitutive over-expression of arginine decarboxylation and putrescine accumulation.

  5. Minimal Power Latch for Single-Slope ADCs

    NASA Technical Reports Server (NTRS)

    Hancock, Bruce R. (Inventor)

    2015-01-01

    A latch circuit that uses two interoperating latches. The latch circuit has the beneficial feature that it switches only a single time during a measurement that uses a stair step or ramp function as an input signal in an analog to digital converter. This feature minimizes the amount of power that is consumed in the latch and also minimizes the amount of high frequency noise that is generated by the latch. An application using a plurality of such latch circuits in a parallel decoding ADC for use in an image sensor is given as an example.

  6. Using clinically acquired MRI to construct age-specific ADC atlases: Quantifying spatiotemporal ADC changes from birth to 6-year old.

    PubMed

    Ou, Yangming; Zöllei, Lilla; Retzepi, Kallirroi; Castro, Victor; Bates, Sara V; Pieper, Steve; Andriole, Katherine P; Murphy, Shawn N; Gollub, Randy L; Grant, Patricia Ellen

    2017-03-31

    Diffusion imaging is critical for detecting acute brain injury. However, normal apparent diffusion coefficient (ADC) maps change rapidly in early childhood, making abnormality detection difficult. In this article, we explored clinical PACS and electronic healthcare records (EHR) to create age-specific ADC atlases for clinical radiology reference. Using the EHR and three rounds of multiexpert reviews, we found ADC maps from 201 children 0-6 years of age scanned between 2006 and 2013 who had brain MRIs with no reported abnormalities and normal clinical evaluations 2+ years later. These images were grouped in 10 age bins, densely sampling the first 1 year of life (5 bins, including neonates and 4 quarters) and representing the 1-6 year age range (an age bin per year). Unbiased group-wise registration was used to construct ADC atlases for 10 age bins. We used the atlases to quantify (a) cross-sectional normative ADC variations; (b) spatiotemporal heterogeneous ADC changes; and (c) spatiotemporal heterogeneous volumetric changes. The quantified age-specific whole-brain and region-wise ADC values were compared to those from age-matched individual subjects in our study and in multiple existing independent studies. The significance of this study is that we have shown that clinically acquired images can be used to construct normative age-specific atlases. These first of their kind age-specific normative ADC atlases quantitatively characterize changes of myelination-related water diffusion in the first 6 years of life. The quantified voxel-wise spatiotemporal ADC variations provide standard references to assist radiologists toward more objective interpretation of abnormalities in clinical images. Our atlases are available at https://www.nitrc.org/projects/mgh_adcatlases. Hum Brain Mapp, 2017. © 2017 Wiley Periodicals, Inc.

  7. Ornithine decarboxylase activity and: [125I]iododeoxyuridine incorporation in rat prostate.

    PubMed Central

    Fuller, D J; Donaldson, L J; Thomas, G H

    1975-01-01

    The relationship between ornithine decarboxylase activity and [125I]iododexyuridine incorporation was studied in prostates from castrated rats (aged 5, 26 and 80 weeks) injected daily with testosterone for up to 10 days. The results suggest that ornithine decarboxylase activity is a parameter of secretory activity, rather than growth, in the ventral prostate. In the dorsolateral prostate, ornithine decarboxylase activity tends to parallel [125I]iododeoxyuridine incorporation. PMID:1212206

  8. Evaluation of commercial ADC radiation tolerance for accelerator experiments

    NASA Astrophysics Data System (ADS)

    Chen, K.; Chen, H.; Kierstead, J.; Takai, H.; Rescia, S.; Hu, X.; Xu, H.; Mead, J.; Lanni, F.; Minelli, M.

    2015-08-01

    Electronic components used in high energy physics experiments are subjected to a radiation background composed of high energy hadrons, mesons and photons. These particles can induce permanent and transient effects that affect the normal device operation. Ionizing dose and displacement damage can cause chronic damage which disable the device permanently. Transient effects or single event effects are in general recoverable with time intervals that depend on the nature of the failure. The magnitude of these effects is technology dependent with feature size being one of the key parameters. Analog to digital converters are components that are frequently used in detector front end electronics, generally placed as close as possible to the sensing elements to maximize signal fidelity. We report on the development of a technique for testing analog to digital converters for radiation effects, in particular for single event effects. A total of seventeen commercial ADCs were evaluated for ionizing dose tolerance and extensive SEU measurements performed on a twelve and fourteen bit ADCs. Mitigation strategies for single event effects (SEE) are discussed for their use in the large hadron collider environment.

  9. An 18 bit 50 kHz ADC for low earth orbit

    NASA Technical Reports Server (NTRS)

    Thelen, Donald C.

    1992-01-01

    A fourth order incremental analog to digital converter (ADC) is proposed which performs 18 bit conversions at a 50 kHz rate on sampled and held data. A new self calibration scheme is presented which eases the matching requirements of capacitors, and the performance of the operational amplifiers in the ADC by changing coefficients in the digital postprocessing.

  10. Optimization of Tubulysin Antibody-Drug Conjugates: A Case Study in Addressing ADC Metabolism.

    PubMed

    Tumey, L Nathan; Leverett, Carolyn A; Vetelino, Beth; Li, Fengping; Rago, Brian; Han, Xiaogang; Loganzo, Frank; Musto, Sylvia; Bai, Guoyun; Sukuru, Sai Chetan K; Graziani, Edmund I; Puthenveetil, Sujiet; Casavant, Jeffrey; Ratnayake, Anokha; Marquette, Kimberly; Hudson, Sarah; Doppalapudi, Venkata Ramana; Stock, Joseph; Tchistiakova, Lioudmila; Bessire, Andrew J; Clark, Tracey; Lucas, Judy; Hosselet, Christine; O'Donnell, Christopher J; Subramanyam, Chakrapani

    2016-11-10

    As part of our efforts to develop new classes of tubulin inhibitor payloads for antibody-drug conjugate (ADC) programs, we developed a tubulysin ADC that demonstrated excellent in vitro activity but suffered from rapid metabolism of a critical acetate ester. A two-pronged strategy was employed to address this metabolism. First, the hydrolytically labile ester was replaced by a carbamate functional group resulting in a more stable ADC that retained potency in cellular assays. Second, site-specific conjugation was employed in order to design ADCs with reduced metabolic liabilities. Using the later approach, we were able to identify a conjugate at the 334C position of the heavy chain that resulted in an ADC with considerably reduced metabolism and improved efficacy. The examples discussed herein provide one of the clearest demonstrations to-date that site of conjugation can play a critical role in addressing metabolic and PK liabilities of an ADC. Moreover, a clear correlation was identified between the hydrophobicity of an ADC and its susceptibility to metabolic enzymes. Importantly, this study demonstrates that traditional medicinal chemistry strategies can be effectively applied to ADC programs.

  11. CHOUGH: petite ADC for a high-order adaptive optics system

    NASA Astrophysics Data System (ADS)

    Hölck, Daniel; Bharmal, Nazim Ali; Black, Martin; Henry, David M.; Myers, Richard M.

    2016-07-01

    We discuss the design of a 50mm diameter Atmospheric Dispersion Corrector (ADC) for The CANARY-Hosted Upgrade for High-Order Adaptive Optics (CHOUGH). Usually to avoid pupil actuator-lenslet array mismatch, the ADC is Customarily placed very close to the pupil plane. This design aims to achieve a non-pupil conjugated ADC suitable to be located in any place inside the collimated beam path, this is due to the restrictions given by CHOUGH optical relay. The ADC also needs to satisfy the very small pupil shift requirement, for pupil stability. The ADC is of the Amici prism type, made up of two plates of cemented double prisms. The two plates counter rotate correcting for the different Zenith angles, from the Zenith up to 60°.

  12. Linearity analysis of single-ended SAR ADC with split capacitive DAC

    NASA Astrophysics Data System (ADS)

    Osipov, Dmitry; Malankin, Evgeny; Shumikhin, Vitaly

    2016-10-01

    This paper proposes the design of a 6-bit single-ended SAR ADC with a variable sampling rate at a maximum achievable speed of 50 MS/s. The SAR ADC utilizes the split capacitor array DAC with a non-conventional split-capacitor value. The influence of switches in the capacitive DAC on the ADC's non-linearity is analysed. According to the fulfilled analysis the recommendations for switches and capacitor array dimensioning are given to provide a minimum differential non-linearity (DNL). At a sampling rate of 50 MS/s, the SAR ADC achieves an ENOB of 5.4 bit at an input signal frequency of 1 MHz and consumes 1.2 mW at a 1.8 V power supply, resulting in an energy efficiency of 568 fJ/conv.-step. The SAR ADC was simulated with parasitics in a standard 180nm CMOS process.

  13. A 12-bit, 40-Ms/s pipelined ADC with an improved operational amplifier

    NASA Astrophysics Data System (ADS)

    Yu, Wang; Haigang, Yang; Tao, Yin; Fei, Liu

    2012-05-01

    This paper proposes a 12-bit, 40-Ms/s pipelined analog-to-digital converter (ADC) with an improved high-gain and wide-bandwidth operational amplifier (opamp). Based on the architecture of the proposed ADC, the non-ideal factors of opamps are first analyzed, which have the significant impact on the ADC's resolution. Then, the compensation techniques of the ADC's opamp are presented to restrain the negative effect introduced by the gain-boosting technique and switched-capacitor common-mode-feedback structure. After analysis and optimization, the ADC implemented in a 0.35 μm standard CMOS process shows a maximum signal-to-noise distortion ratio of 60.5 dB and a spurious-free dynamic range of 74.5 dB, respectively, at a 40 MHz sample clock with over 2 Vpp input range.

  14. Evaluation of a Modified ADC-Based Thermometry Bridge

    NASA Astrophysics Data System (ADS)

    Podgornik, T.; Batagelj, V.; Winkler, G.; Hartl, H.; Drnovsek, J.

    2011-12-01

    This article presents the modification and testing of an ADC-based thermometry bridge. The instrument under investigation is an Anton Paar MKT 50 Millikelvin Thermometer (developed at the IFE, TU-Graz) based on a precision analog-to-digital converter (ADC). During preliminary testing, it was found that the MKT 50 performs better than its declared uncertainty (1 mK equal to 1 ppm when using a 100 Ω PRT) and is comparable to thermometry resistance ratio bridges typically used in secondary thermometry laboratories (with typical uncertainties from 0.1 mK to 1 mK). The modifications to the original bridge were undertaken by the development team of the MKT 50 at the Graz University of Technology, Austria. Measurements and evaluation of the modified instruments were performed at the MIRS/UL-FE/LMK. For the MKT 50 to be used in thermometry laboratories as a reference unit, measuring parameters of the instrument had to be changed. During the first modification, the upper limit of the instrument range was decreased from 400 Ω to 133 Ω, this is a preferred range for standard platinum resistance thermometers (SPRTs). This also meant an increase in the measuring current from 0.5 mA to the more frequently used 1 mA. A modification of the programmable ADC control unit increased the resolution from 24 bit to 27 bit. By adding a switch, the use of an external standard resistor was enabled. After this stage of the modification, the first tests on the instrument were performed. The second stage was aimed at the removal of noise sources. The instrument was prepared in such a way that it only used two input channels, one connected to the SPRT and the other to the standard resistor. Also, the components of the ADC were upgraded to further reduce noise. The elimination of one input channel sped up measurements, making the PC software capable of taking several readings in a shorter time period. All tests were performed in laboratory conditions, where precision AC and DC resistance

  15. Cell biology, physiology and enzymology of phosphatidylserine decarboxylase.

    PubMed

    Di Bartolomeo, Francesca; Wagner, Ariane; Daum, Günther

    2017-01-01

    Phosphatidylethanolamine is one of the most abundant phospholipids whose major amounts are formed by phosphatidylserine decarboxylases (PSD). Here we provide a comprehensive description of different types of PSDs in the different kingdoms of life. In eukaryotes, type I PSDs are mitochondrial enzymes, whereas other PSDs are localized to other cellular compartments. We describe the role of mitochondrial Psd1 proteins, their function, enzymology, biogenesis, assembly into mitochondria and their contribution to phospholipid homeostasis in much detail. We also discuss briefly the cellular physiology and the enzymology of Psd2. This article is part of a Special Issue entitled: Lipids of Mitochondria edited by Guenther Daum.

  16. Resolution of brewers' yeast pyruvate decarboxylase into two isozymes.

    PubMed

    Kuo, D J; Dikdan, G; Jordan, F

    1986-03-05

    A novel purification method was developed for brewers' yeast pyruvate decarboxylase (EC 4.1.1.1) that for the first time resolved the enzyme into two isozymes on DEAE-Sephadex chromatography. The isozymes were found to be distinct according to sodium dodecyl sulfate polyacrylamide gel electrophoresis: the first one to be eluted gave rise to one band, the second to two bands. The isozymes were virtually the same so far as specific activity, KM, inhibition kinetics and irreversible binding properties by the mechanism-based inhibitor (E)-4-(4-chlorophenyl)-2-oxo-3-butenoic acid are concerned. This finding resolves a longstanding controversy concerning the quaternary structure of this enzyme.

  17. Vector-mediated chromosomal integration of the glutamate decarboxylase gene in streptococcus thermophilus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The integrative vector pINTRS was used to transfer glutamate decarboxylase (GAD) activity to Streptococcus thermophilus ST128, thus allowing for the production of '-aminobutyric acid (GABA). In pINTRS, the gene encoding glutamate decarboxylase, gadB, was flanked by DNA fragments homologous to a S. ...

  18. Changes in ADC Caused by Tensile Loading of Rabbit Achilles Tendon: Evidence for Water Transport

    NASA Astrophysics Data System (ADS)

    Han, S.; Gemmell, S. J.; Helmer, K. G.; Grigg, P.; Wellen, J. W.; Hoffman, A. H.; Sotak, C. H.

    2000-06-01

    Water diffusion measurements were performed on rabbit Achilles tendons during static tensile loading and tendons in an unloaded state. The apparent diffusion coefficient (ADC) was measured along two directions: parallel and perpendicular to the long axis of the tendon. Tendons were studied after being prepared in two ways: (a) after being stored frozen in phosphate-buffered saline (PBS) and (b) freshly isolated. Statistically significant directional anisotropy was observed in the ADC in all tendons. The ADC was significantly greater in the direction parallel to the long axis of the tendon than in the perpendicular direction. The anisotropy is attributed to the greater restrictions seen by the water molecules in the perpendicular direction and is consistent with the known geometry of the tendon. Storage in PBS caused tendons to swell. This increased the ADC measured along both directions and reduced the anisotropy. The existence of anisotropy in the ADC was not related to the orientation of the specimen in the magnet. The ADC increased along both directions following the application of a 5-N tensile load; the increase was greatest along the perpendicular axis of the tendon. In order to determine whether load-related changes in the ADC reflected changes in interfibrilar spacing, we used electron microscopy to measure load-related changes in fibril spacing. Load-related changes in fiber spacing could not account for the observed changes in the ADC. The increase in ADC caused by loading was attributed to the extrusion of tendon water into a bulk phase along the outside surface of the tendon. In PBS-stored samples, enough fluid was extruded that it could be visualized. The transient response of the ADC to a 5-N tensile load was also studied. The absolute ADC in both directions increased with loading and recovered to baseline upon unloading. The transient changes in ADC, for both loading and unloading, had a mean time constant of approximately 15 min. The magnitude of

  19. Functionally diverse biotin-dependent enzymes with oxaloacetate decarboxylase activity.

    PubMed

    Lietzan, Adam D; St Maurice, Martin

    2014-02-15

    Biotin-dependent enzymes catalyze carboxylation, decarboxylation and transcarboxylation reactions that participate in the primary metabolism of a wide range of organisms. In all cases, the overall reaction proceeds via two half reactions that take place in physically distinct active sites. In the first half-reaction, a carboxyl group is transferred to the 1-N' of a covalently tethered biotin cofactor. The tethered carboxybiotin intermediate subsequently translocates to a second active site where the carboxyl group is either transferred to an acceptor substrate or, in some bacteria and archaea, is decarboxylated to biotin and CO2 in order to power the export of sodium ions from the cytoplasm. A homologous carboxyltransferase domain is found in three enzymes that catalyze diverse overall reactions: carbon fixation by pyruvate carboxylase, decarboxylation and sodium transport by the biotin-dependent oxaloacetate decarboxylase complex, and transcarboxylation by transcarboxylase from Propionibacterium shermanii. Over the past several years, structural data have emerged which have greatly advanced the mechanistic description of these enzymes. This review assembles a uniform description of the carboxyltransferase domain structure and catalytic mechanism from recent studies of pyruvate carboxylase, oxaloacetate decarboxylase and transcarboxylase, three enzymes that utilize an analogous carboxyltransferase domain to catalyze the biotin-dependent decarboxylation of oxaloacetate.

  20. Tyrosine decarboxylase from Lactobacillus brevis: soluble expression and characterization.

    PubMed

    Zhang, Kai; Ni, Ye

    2014-02-01

    Tyrosine decarboxylase (TDC, EC 4.1.1.25) is an enzyme that catalyzes the decarboxylation of l-tyrosine to produce tyramine and CO2. In this study, a 1881-bp tdc gene from Lactobacillus brevis was cloned and heterologously expressed in Escherichia coli BL21 (DE3). Glucose was discovered to play an important role in the soluble expression of rLbTDC. After optimization, recombinant TDC (rLbTDC) was achieved in excellent solubility and a yield of 224mg rLbTDC/L broth. The C-terminal His-Tagged rLbTDC was one-step purified with 90% recovery. Based on SDS-PAGE and gel filtration analysis, rLbTDC is a dimer composed of two identical subunits of approximately 70kDa. Using l-tyrosine as substrate, the specific activity of rLbTDC was determined to be 133.5U/mg in the presence of 0.2mM pyridoxal-5'-phosphate at 40°C and pH 5.0. The Km and Vmax values of rLbTDC were 0.59mM and 147.1μmolmin(-1)mg(-1), respectively. In addition to l-tyrosine, rLbTDC also exhibited decarboxylase activity towards l-DOPA. This study has demonstrated, for the first time, the soluble expression of tdc gene from L. brevis in heterologous host.

  1. Crystal structure of pyruvate decarboxylase from Zymobacter palmae

    PubMed Central

    Buddrus, Lisa; Andrews, Emma S. V.; Leak, David J.; Danson, Michael J.; Arcus, Vickery L.; Crennell, Susan J.

    2016-01-01

    Pyruvate decarboxylase (PDC; EC 4.1.1.1) is a thiamine pyrophosphate- and Mg2+ ion-dependent enzyme that catalyses the non-oxidative decarboxylation of pyruvate to acetaldehyde and carbon dioxide. It is rare in bacteria, but is a key enzyme in homofermentative metabolism, where ethanol is the major product. Here, the previously unreported crystal structure of the bacterial pyruvate decarboxylase from Zymobacter palmae is presented. The crystals were shown to diffract to 2.15 Å resolution. They belonged to space group P21, with unit-cell parameters a = 204.56, b = 177.39, c = 244.55 Å and R r.i.m. = 0.175 (0.714 in the highest resolution bin). The structure was solved by molecular replacement using PDB entry 2vbi as a model and the final R values were R work = 0.186 (0.271 in the highest resolution bin) and R free = 0.220 (0.300 in the highest resolution bin). Each of the six tetramers is a dimer of dimers, with each monomer sharing its thiamine pyrophosphate across the dimer interface, and some contain ethylene glycol mimicking the substrate pyruvate in the active site. Comparison with other bacterial PDCs shows a correlation of higher thermostability with greater tetramer interface area and number of interactions. PMID:27599861

  2. A robust and simple two-mode digital calibration technique for pipelined ADC

    NASA Astrophysics Data System (ADS)

    Xiumei, Yin; Nan, Zhao; Bomeh Kobenge, Sekedi; Huazhong, Yang

    2011-03-01

    This paper presents a two-mode digital calibration technique for pipelined analog-to-digital converters (ADC). The proposed calibration eliminates the errors of residual difference voltage induced by capacitor mismatch of pseudorandom (PN) sequence injection capacitors at the ADC initialization, while applies digital background calibration to continuously compensate the interstage gain errors in ADC normal operation. The presented technique not only reduces the complexity of analog circuit by eliminating the implementation of PN sequence with accurate amplitude in analog domain, but also improves the performance of digital background calibration by minimizing the sensitivity of calibration accuracy to sub-ADC errors. The use of opamps with low DC gains in normal operation makes the proposed design more compatible with future nanometer CMOS technology. The prototype of a 12-bit 40-MS/s pipelined ADC with the two-mode digital calibration is implemented in 0.18-μm CMOS process. Adopting a simple telescopic opamp with a DC gain of 58-dB in the first stage, the measured SFDR and SNDR within the first Nyquist zone reach 80-dB and 66-dB, respectively. With the calibration, the maximum integral nonlinearity (INL) of the ADC reduces from 4.75-LSB to 0.65-LSB, while the ADC core consumes 82-mW at 3.3-V power supply.

  3. ADCS Prevention Instrument Project: overview and initial results.

    PubMed

    Ferris, Steven H; Aisen, Paul S; Cummings, Jeffrey; Galasko, Douglas; Salmon, David P; Schneider, Lon; Sano, Mary; Whitehouse, Peter J; Edland, Steven; Thal, Leon J

    2006-01-01

    One objective of the Alzheimer's Disease Cooperative Study (ADCS) is to develop new or improved instruments and assessment methods for evaluating treatment efficacy in Alzheimer disease (AD) clinical trials. The ADCS Instrument Committee has previously helped to define the state of the art in assessment for AD and Mild Cognitive Impairment clinical trials. We are now entering an exciting era of primary prevention trials to evaluate promising treatments that may delay disease onset and there is a need to develop appropriate instruments for these trials. The ADCS instrument committee has undertaken a project to develop instruments for prevention studies that assess domains known to be important in AD. Prevention trials are long and require large numbers of subjects, making them costly and requiring a high burden of participation for subjects. The current study focused on developing instruments that can be completed at home and in the clinic. The instruments are being evaluated in a cohort of nondemented elderly participating in a 4-year longitudinal study that simulates the design of a primary prevention trial. This report describes the design, baseline characteristics, and some longitudinal outcomes of the study cohort through the completion of the first 2 years of follow-up. We also describe the assessment domains to be measured with our new experimental instruments. This study recruited 644 subjects, 75 years of age and older. Participation in a "book club" that provided free books of interest to elders was offered as a recruitment incentive. Approximately 23% had some mild cognitive symptoms consistent with a Clinical Dementia Rating of 0.5. All subjects received a standardized in-clinic evaluation at baseline, which is repeated annually for 4 years to identify cases suspected of developing dementia and to measure longitudinal change on established clinical assessments. Subjects completed a set of self-administered experimental instruments at home or in the

  4. ADCS controllers comparison for small satellitess in Low Earth Orbit

    NASA Astrophysics Data System (ADS)

    Calvo, Daniel; Laverón-Simavilla, Ana; Lapuerta, Victoria

    2016-07-01

    Fuzzy logic controllers are flexible and simple, suitable for small satellites Attitude Determination and Control Subsystems (ADCS). In a previous work, a tailored Fuzzy controller was designed for a nanosatellite. Its performance and efficiency were compared with a traditional Proportional Integrative Derivative (PID) controller within the same specific mission. The orbit height varied along the mission from injection at around 380 km down to 200 km height, and the mission required pointing accuracy over the whole time. Due to both, the requirements imposed by such a low orbit, and the limitations in the power available for the attitude control, an efficient ADCS is required. Both methodologies, fuzzy and PID, were fine-tuned using an automated procedure to grant maximum efficiency with fixed performances. The simulations showed that the Fuzzy controller is much more efficient (up to 65% less power required) in single manoeuvres, achieving similar, or even better, precision than the PID. The accuracy and efficiency improvement of the Fuzzy controller increase with orbit height because the environmental disturbances decrease, approaching the ideal scenario. However, the controllers are meant to be used in a vast range of situations and configurations which exceed those used in the calibration process carried out in the previous work. To assess the suitability and performance of both controllers in a wider framework, parametric and statistical methods have been applied using the Monte Carlo technique. Several parameters have been modified randomly at the beginning of each simulation: the moments of inertia of the whole satellite and of the momentum wheel, the residual magnetic dipole and the initial conditions of the test. These parameters have been chosen because they are the main source of uncertainty during the design phase. The variables used for the analysis are the error (critical for science) and the operation cost (which impacts the mission lifetime and

  5. Uncovering the Lactobacillus plantarum WCFS1 Gallate Decarboxylase Involved in Tannin Degradation

    PubMed Central

    Jiménez, Natalia; Curiel, José Antonio; Reverón, Inés; de las Rivas, Blanca

    2013-01-01

    Lactobacillus plantarum is a lactic acid bacterium able to degrade tannins by the subsequent action of tannase and gallate decarboxylase enzymes. The gene encoding tannase had previously been identified, whereas the gene encoding gallate decarboxylase is unknown. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of gallic-acid induced L. plantarum extracts showed a 54-kDa protein which was absent in the uninduced cells. This protein was identified as Lp_2945, putatively annotated UbiD. Homology searches identified ubiD-like genes located within three-gene operons which encoded the three subunits of nonoxidative aromatic acid decarboxylases. L. plantarum is the only bacterium in which the lpdC (lp_2945) gene and the lpdB and lpdD (lp_0271 and lp_0272) genes are separated in the chromosome. Combination of extracts from recombinant Escherichia coli cells expressing the lpdB, lpdC, and lpdC genes demonstrated that LpdC is the only protein required to yield gallate decarboxylase activity. However, the disruption of these genes in L. plantarum revealed that the lpdB and lpdC gene products are essential for gallate decarboxylase activity. Similar to L. plantarum tannase, which exhibited activity only in esters derived from gallic and protocatechuic acids, purified His6-LpdC protein from E. coli showed decarboxylase activity against gallic and protocatechuic acids. In contrast to the tannase activity, gallate decarboxylase activity is widely present among lactic acid bacteria. This study constitutes the first genetic characterization of a gallate decarboxylase enzyme and provides new insights into the role of the different subunits of bacterial nonoxidative aromatic acid decarboxylases. PMID:23645198

  6. Evaluation of commercial ADC radiation tolerance for accelerator experiments

    DOE PAGES

    Chen, K.; Chen, H.; Kierstead, J.; ...

    2015-08-17

    Electronic components used in high energy physics experiments are subjected to a radiation background composed of high energy hadrons, mesons and photons. These particles can induce permanent and transient effects that affect the normal device operation. Ionizing dose and displacement damage can cause chronic damage which disable the device permanently. Transient effects or single event effects are in general recoverable with time intervals that depend on the nature of the failure. The magnitude of these effects is technology dependent with feature size being one of the key parameters. Analog to digital converters are components that are frequently used in detectormore » front end electronics, generally placed as close as possible to the sensing elements to maximize signal fidelity. We report on radiation effects tests conducted on 17 commercially available analog to digital converters and extensive single event effect measurements on specific twelve and fourteen bit ADCs that presented high tolerance to ionizing dose. We discuss mitigation strategies for single event effects (SEE) for their use in the large hadron collider environment.« less

  7. Evaluation of commercial ADC radiation tolerance for accelerator experiments

    SciTech Connect

    Chen, K.; Chen, H.; Kierstead, J.; Takai, H.; Rescia, S.; Hu, X.; Xu, H.; Mead, J.; Lanni, F.; Minelli, M.

    2015-08-17

    Electronic components used in high energy physics experiments are subjected to a radiation background composed of high energy hadrons, mesons and photons. These particles can induce permanent and transient effects that affect the normal device operation. Ionizing dose and displacement damage can cause chronic damage which disable the device permanently. Transient effects or single event effects are in general recoverable with time intervals that depend on the nature of the failure. The magnitude of these effects is technology dependent with feature size being one of the key parameters. Analog to digital converters are components that are frequently used in detector front end electronics, generally placed as close as possible to the sensing elements to maximize signal fidelity. We report on radiation effects tests conducted on 17 commercially available analog to digital converters and extensive single event effect measurements on specific twelve and fourteen bit ADCs that presented high tolerance to ionizing dose. We discuss mitigation strategies for single event effects (SEE) for their use in the large hadron collider environment.

  8. Characterizing the Performance of a High-Speed ADC for the Sma Digital Backend

    NASA Astrophysics Data System (ADS)

    Patel, N. A.; Wilson, R. W.; Primiani, R.; Weintroub, J.; Test, J.; Young, K.

    We report on tests of a 5 Gs/s analog-to-digital converter (ADC) used in the new Submillimeter Array (SMA) Digital Backend (DBE). The ADC is e2v EV8AQ160, with 8-bit resolution and 4 interleaved cores, operated in single-channel mode. We measured the frequency response, Signal to Noise and Distortion (SINAD), Spurious Free Dynamic Range (SFDR), Noise Power Ratio and intermodulation distortion over the bandwidth of 2.25 GHz. The performance of this ADC is found to be adequate for our application in the SMA DBE. We describe the procedure of aligning the four cores for adjustments of offset, gain and phase parameters which improve the performance of the ADC, particularly in SINAD and SFDR.

  9. Is anoxic depolarisation associated with an ADC threshold? A Markov chain Monte Carlo analysis.

    PubMed

    King, Martin D; Crowder, Martin J; Hand, David J; Harris, Neil G; Williams, Stephen R; Obrenovitch, Tihomir P; Gadian, David G

    2005-12-01

    A Bayesian nonlinear hierarchical random coefficients model was used in a reanalysis of a previously published longitudinal study of the extracellular direct current (DC)-potential and apparent diffusion coefficient (ADC) responses to focal ischaemia. The main purpose was to examine the data for evidence of an ADC threshold for anoxic depolarisation. A Markov chain Monte Carlo simulation approach was adopted. The Metropolis algorithm was used to generate three parallel Markov chains and thus obtain a sampled posterior probability distribution for each of the DC-potential and ADC model parameters, together with a number of derived parameters. The latter were used in a subsequent threshold analysis. The analysis provided no evidence indicating a consistent and reproducible ADC threshold for anoxic depolarisation.

  10. [Inhibitory effect of essential oils, food additives, peracetic acid and detergents on bacterial histidine decarboxylase].

    PubMed

    Kamii, Eri; Terada, Gaku; Akiyama, Jyunki; Isshiki, Kenji

    2011-01-01

    The aim of this study is to examine whether various essential oils, food additives, peracetic acid and detergents inhibit bacterial histidine decarboxylase. Crude extract of Morganella morganii NBRC3848 was prepared and incubated with various agents. Histidine decarboxylase activity was significantly inhibited (p<0.05) by 26 of 45 compounds tested. Among the 26 agents, sodium hypochlorite completely decomposed both histidine and histamine, while peracetic acid caused slight decomposition. Histidine and histamine were stable in the presence of the other 24 agents. These results indicated that 25 of the agents examined were inhibitors of histidine decarboxylase.

  11. Glutamic acid decarboxylase autoimmunity in Batten disease and other disorders.

    PubMed

    Pearce, David A; Atkinson, Mark; Tagle, Danilo A

    2004-12-14

    Degenerative diseases of the CNS, such as stiff-person syndrome (SPS), progressive cerebellar ataxia, and Rasmussen encephalitis, have been characterized by the presence of autoantibodies. Recent findings in individuals with Batten disease and in animal models for the disorder indicate that this condition may be associated with autoantibodies against glutamic acid decarboxylase (GAD), an enzyme that converts the excitatory neurotransmitter glutamate to the inhibitory neurotransmitter gamma-aminobutyric acid (GABA). Anti-GAD autoantibodies could result in excess excitatory neurotransmitters, leading to the seizures and other symptoms observed in patients with Batten disease. The pathogenic potential of GAD autoantibodies is examined in light of what is known for other autoimmune disorders, such as multiple sclerosis, SPS, Rasmussen encephalitis, and type 1 diabetes, and may have radical implications for diagnosis and management of Batten disease.

  12. Levodopa combined with peripheral decarboxylase inhibition in Parkinson's disease

    PubMed Central

    Barbeau, André; Mars, Harold; Botez, Mihai I.; Joubert, Marie

    1972-01-01

    The authors report their experience, over a 26-month period, in the management of 60 parkinsonian patients with the combination of levodopa and an inhibitor of peripheral dopa-decarboxylase, Ro 4-4602. This approach to Parkinson's disease is useful, safe, and at least as effective as levodopa alone. To date there have been no recognizable toxic effects attributable to Ro 4-4602. This agent appears to prolong the duration of action of levodopa, smoothing out its therapeutic effects. The percentage of patients obtaining a very good and excellent response is slightly increased. There is a possible diminution in the late-occurring bradykinetic and hypotonic freezing episodes. Nausea and cardiac arrhythmias are lessened, as are the incidence and severity of hypotension. Abnormal involuntary movements remain the limiting adverse side effect. PMID:5034697

  13. An endosymbiont positively modulates ornithine decarboxylase in host trypanosomatids

    SciTech Connect

    Frossard, Mariana Lins; Seabra, Sergio Henrique; Matta, Renato Augusto da; Souza, Wanderley de; Garcia de Mello, Fernando; Motta, Maria Cristina Machado . E-mail: motta@biof.ufrj.br

    2006-05-05

    Summary: Some trypanosomatids, such as Crithidia deanei, are endosymbiont-containing species. Aposymbiotic strains are obtained after antibiotic treatment, revealing interesting aspects of this symbiotic association. Ornithine decarboxylase (ODC) promotes polyamine biosynthesis and contributes to cell proliferation. Here, we show that ODC activity is higher in endosymbiont-bearing trypanosomatids than in aposymbiotic cells, but isolated endosymbionts did not display this enzyme activity. Intriguingly, expressed levels of ODC were similar in both strains, suggesting that ODC is positively modulated in endosymbiont-bearing cells. When the aposymbiotic strain was grown in conditioned medium, obtained after cultivation of the endosymbiont-bearing strain, cellular proliferation as well as ODC activity and localization were similar to that observed in the endosymbiont-containing trypanosomatids. Furthermore, dialyzed-heated medium and trypsin treatment reduced ODC activity of the aposymbiont strain. Taken together, these data indicate that the endosymbiont can enhance the protozoan ODC activity by providing factors of protein nature, which increase the host polyamine metabolism.

  14. Altered subcellular localization of ornithine decarboxylase in Alzheimer's disease brain

    SciTech Connect

    Nilsson, Tatjana . E-mail: Tatjana.Nilsson@ki.se; Bogdanovic, Nenad; Volkman, Inga; Winblad, Bengt; Folkesson, Ronnie; Benedikz, Eirikur

    2006-06-02

    The amyloid precursor protein can through ligand-mimicking induce expression of ornithine decarboxylase (ODC), the initial and rate-limiting enzyme in polyamine biosynthesis. We report here the regional distribution and cellular localization of ODC immunoreactivity in Alzheimer's disease (AD) brains. In frontal cortex and hippocampus of control cases, the most pronounced ODC immunoreactivity was found in the nucleus. In possible and definite AD the immunoreactivity had shifted to the cytoplasm. In cerebellum of control cases, ODC staining was found in a small portion of Purkinje cells, mostly in the nucleus. In AD, both possible and definite, the number of stained Purkinje cells increased significantly and immunoreactivity was shifted to the cytoplasm, even though it was still prominent in the nucleus. In conclusion, our study reveals an early shift of the ODC immunoreactivity in AD from the nuclear compartment towards the cytoplasm.

  15. Mouse ornithine decarboxylase gene: cloning, structure, and expression.

    PubMed Central

    Brabant, M; McConlogue, L; van Daalen Wetters, T; Coffino, P

    1988-01-01

    We used molecular cloning to isolate a functional gene for mouse ornithine decarboxylase (OrnDCase; L-ornithine carboxy-lyase, EC 4.1.1.17) from a cell line in which that gene had been selectively amplified. The position of the 5' terminus of the mRNA was identified, and the coding sequence was shown to be preceded by a 312- or 313-nucleotide (nt) untranslated leader. The latter is highly G + C rich, particularly in its 5'-most portion. The leader can be anticipated to have extensive and stable secondary structure. The transcription unit of the gene is of relatively small size, approximately equal to 6.2 kilobases (kb) from the start site to the proximal site of polyadenylylation. Sequence analysis of DNA near the transcription start position demonstrated the presence of a "TATA" box, but no "CAAT" box. Functional properties of the cloned gene were tested by transfecting it into cultured cells. Expression of the putative full-length gene efficiently conferred ornithine decarboxylase activity on recipient mutant cells deficient in that activity. To assess the function and strength of the OrnDCase promoter region and to delimit its boundaries, we used a transient expression assay. Upstream of a bacterial chloramphenicol acetyltransferase gene was placed a portion of the OrnDCase gene, including the presumed promoter region, spanning a region from approximately equal to 3.0 kb 5' of the site of transcription initiation to the first 250 nt of the transcript. When expressed in mouse NIH 3T3 cells, this OrnDCase genomic element was comparable in strength to the Rous sarcoma virus long terminal repeat promoter. A similar construct, truncated so as to retain only 264 base pairs of the OrnDCase gene 5' to the site of transcription start, yielded undiminished levels of expression. Images PMID:3353375

  16. A FASTBUS flash ADC system for the Mark II vertex chamber

    SciTech Connect

    Barker, L.

    1988-10-01

    This is a description of a flash ADC system built for the Mark II experiment at the Stanford Linear Accelerator Center (SLAC). This system was designed for use in the experiment's vertex chamber where signals could occur over a relatively long time, approximately 10 microseconds. This long time, coupled with fast cable amplifiers, necessitated an alternate design approach than was used with a dE/dX FASTBUS flash ADC design. 1 ref., 6 figs.

  17. Method of correcting of the tracking ADC with weight redundancy conversion characteristic

    NASA Astrophysics Data System (ADS)

    Azarov, Oleksiy D.; Dudnyk, Oleksandr V.; Kaduk, Oleksandr V.; Smolarz, Andrzej; Burlibay, Aron

    2015-12-01

    The methods of correction of the saltatory conversion characteristics of tracking ADC with weight redundancy, based on inaccurate analog elements, were analyzed. It is shown that the multiplicity of numbers presentation in notation systems with weight redundancy allows to reduce the errors of the conversion characteristic of a DAC and to simplify the correction of the conversion characteristic of the ADC containing such DAC using the boundary code combinations. The criterion was defined and the effectiveness of the proposed approach was estimated.

  18. ADC common noise correction and zero suppression in the PIBETA detector

    NASA Astrophysics Data System (ADS)

    Frlež, E.; Počanić, D.; Ritt, S.

    2001-05-01

    We describe a simple procedure for reducing Analog-to-Digital Converter (ADC) common noise in modular detectors that does not require additional hardware. A method using detector noise groups should work well for modular particle detectors such as segmented electromagnetic calorimeters, plastic scintillator hodoscopes, cathode strip wire chambers, segmented active targets, and the like. We demonstrate a "second pedestal noise correction" method by comparing representative ADC pedestal spectra for various elements of the PIBETA detector before and after the applied correction.

  19. Spatially distributed successive approximation register (SDSAR) photonic ADCs based on phase-domain quantization.

    PubMed

    Nazarathy, Moshe; Shaham, Oded

    2012-03-26

    We explore photonic ADC architectures based on encoding voltage-under-test into phase. The first step is to identify two basic optical building blocks: the optical phase comparator (1-bit ADC), based on interferometric comparison of phases in the well-known balanced photo-detection configuration, and the optical 1-bit DAC, namely electro-optic modulation with a bipolar electrical pulse. Equipped with these fundamental building blocks, we proceed to systematically port and adapt known ADC quantization architectures to photonic ADC, conceiving a hybrid between the Successive Approximation Register (SAR) and the Pipeline classic ADC architectures, referred to here as Spatially Distributed SAR (SDSAR). This novel photonic ADC, constructed out of B 1-bit ADCs and B-2 1-bit DACs, with B the number of bits, is not equivalent to any of the previous photonic ADCs in the literature, but appears superior to prior schemes in both optical power efficiency and electro-optic modulation complexity. We derive upper bounds on resolution, Effective Number of Bits (ENOB) performance as a function of average optical power for the new SDSAR device, developing analytic and numeric Monte-Carlo statistical models, comprising quantization, shot, thermal and DAC voltage noise sources. Our findings indicate that SDSAR is limited to ~11.5 ENOBs, assuming state-of-the-art mode-locked-lasers providing ~250 mW of average power (assuming ~7 dB excess losses). However, this upper bound is not tight, due to various physical impairments. In particular, the mode locked laser jitter is shown to have negligible impact on overall performance for RMS jitter < 20 fsec.

  20. 64-Channel, 5 GSPS ADC Module with Switched Capacitor Arrays

    NASA Astrophysics Data System (ADS)

    Bogdan, M.; Huan, H.; Wakely, S.

    2013-08-01

    We present a 5 GSPS ADC/Data processing module with up to 64 channels and 2048 cells per channel, designed for fast-sampling, front-end applications. This is a 6U VME board that incorporates 16 pieces DRS4 (http://drs.web.psi.ch, [1]) Switched Capacitor Array chips developed at Paul Scherrer Institut, Switzerland. The 16 DRS4 chips are grouped in four independent input blocks. A block, with a geometric size of 43×120 mm, has four pieces DRS4 chips, four pieces AD9222 converters, and one Altera Stratix III FPGA. Each DRS4 chip has eight channels and each channel has 1024 sampling cells, which can be daisy-chained for larger sampling depth. This feature allows for a great level of flexibility in choosing the number of channels relative to capacitor array size, for a particular application. The first prototype Printed Circuit Board (PCB) was designed for a sampling depth of 2048 cells and 16 channels in a 42 mm wide block, i.e. 64 channels for the 6U VME board. This compact form factor allows for these input blocks to be used as front-end electronics for the Cherenkov Telescope Array (CTA) cameras. In this VME board, the four blocks are fully independent and can run each in different modes without any conflict. A global FPGA, also a Stratix III device, provides control and interfacing. The module can run with a local oscillator or with input system clocks in the range of 20-550 MHz. The front panel is fitted with a 2.5 Gbps serial link transceiver.

  1. Coenzyme A biosynthesis: steric course of 4'-phosphopantothenoyl-L-cysteine decarboxylase.

    PubMed

    Aberhart, D J; Ghoshal, P K; Cotting, J A; Russell, D J

    1985-12-03

    4'-Phosphopantothenoyl-L-cysteine decarboxylase (PPC decarboxylase) was partially purified from rat liver. 4'-Phosphopantothenoyl[2-2H1]-L-cysteine was synthesized and converted by PPC decarboxylase to 4'-phosphol[1-2H1]pantetheine. The product was degraded by reduction with Raney nickel followed by acidic hydrolysis to [1-2H1]ethylamine. The latter was converted to the (-)-camphanamide derivative, NMR studies of which revealed that the deuterium was located in the pro-1S position. Also, unlabeled 4'-phosphopantothenoyl-L-cysteine was incubated with PPC decarboxylase in D2O, giving, after degradation, the (-)-camphanamide of (1R)-[1-2H1]ethylamine. The results show that the decarboxylation takes place with retention of configuration. These results are discussed in terms of possible mechanisms for the decarboxylation.

  2. A Liquid-Based Colorimetric Assay of Lysine Decarboxylase and Its Application to Enzymatic Assay.

    PubMed

    Kim, Yong Hyun; Sathiyanarayanan, Ganesan; Kim, Hyun Joong; Bhatia, Shashi Kant; Seo, Hyung-Min; Kim, Jung-Ho; Song, Hun-Seok; Kim, Yun-Gon; Park, Kyungmoon; Yang, Yung-Hun

    2015-12-28

    A liquid-based colorimetric assay using a pH indicator was introduced for high-throughput monitoring of lysine decarboxylase activity. The assay is based on the color change of bromocresol purple, measured at 595 nm in liquid reaction mixture, due to an increase of pH by the production of cadaverine. Bromocresol purple was selected as the indicator because it has higher sensitivity than bromothymol blue and pheonol red within a broad range and shows good linearity within the applied pH. We applied this for simple determination of lysine decarboxylase reusability using 96-well plates, and optimization of conditions for enzyme overexpression with different concentrations of IPTG on lysine decarboxylase. This assay is expected to be applied for monitoring and quantifying the liquid-based enzyme reaction in biotransformation of decarboxylase in a high-throughput way.

  3. Structural Basis of the Substrate Specificity and Enzyme Catalysis of a Papaver somniferum Tyrosine Decarboxylase.

    PubMed

    Guan, Huai; Song, Shuaibao; Robinson, Howard; Liang, Jing; Ding, Haizhen; Li, Jianyong; Han, Qian

    2017-01-01

    Tyrosine decarboxylase (TyDC), a type II pyridoxal 5'-phosphate decarboxylase, catalyzes the decarboxylation of tyrosine. Due to a generally high sequence identity to other aromatic amino acid decarboxylases (AAADs), primary sequence information is not enough to understand substrate specificities with structural information. In this study, we selected a typical TyDC from Papaver somniferum as a model to study the structural basis of AAAD substrate specificities. Analysis of the native P. somniferum TyDC crystal structure and subsequent molecular docking and dynamics simulation provide some structural bases that explain substrate specificity for tyrosine. The result confirmed the previous proposed mechanism for the enzyme selectivity of indolic and phenolic substrates. Additionally, this study yields the first crystal structure for a plant type II pyridoxal-5'-phosphate decarboxylase.

  4. Structural Basis of the Substrate Specificity and Enzyme Catalysis of a Papaver somniferum Tyrosine Decarboxylase

    PubMed Central

    Guan, Huai; Song, Shuaibao; Robinson, Howard; Liang, Jing; Ding, Haizhen; Li, Jianyong; Han, Qian

    2017-01-01

    Tyrosine decarboxylase (TyDC), a type II pyridoxal 5′-phosphate decarboxylase, catalyzes the decarboxylation of tyrosine. Due to a generally high sequence identity to other aromatic amino acid decarboxylases (AAADs), primary sequence information is not enough to understand substrate specificities with structural information. In this study, we selected a typical TyDC from Papaver somniferum as a model to study the structural basis of AAAD substrate specificities. Analysis of the native P. somniferum TyDC crystal structure and subsequent molecular docking and dynamics simulation provide some structural bases that explain substrate specificity for tyrosine. The result confirmed the previous proposed mechanism for the enzyme selectivity of indolic and phenolic substrates. Additionally, this study yields the first crystal structure for a plant type II pyridoxal-5'-phosphate decarboxylase. PMID:28232911

  5. Glutamic acid decarboxylase isoform distribution in transgenic mouse septum: an anti-GFP immunofluorescence study.

    PubMed

    Verimli, Ural; Sehirli, Umit S

    2016-09-01

    The septum is a basal forebrain region located between the lateral ventricles in rodents. It consists of lateral and medial divisions. Medial septal projections regulate hippocampal theta rhythm whereas lateral septal projections are involved in processes such as affective functions, memory formation, and behavioral responses. Gamma-aminobutyric acidergic neurons of the septal region possess the 65 and 67 isoforms of the enzyme glutamic acid decarboxylase. Although data on the glutamic acid decarboxylase isoform distribution in the septal region generally appears to indicate glutamic acid decarboxylase 67 dominance, different studies have given inconsistent results in this regard. The aim of this study was therefore to obtain information on the distributions of both of these glutamic acid decarboxylase isoforms in the septal region in transgenic mice. Two animal groups of glutamic acid decarboxylase-green fluorescent protein knock-in transgenic mice were utilized in the experiment. Brain sections from the region were taken for anti-green fluorescent protein immunohistochemistry in order to obtain estimated quantitative data on the number of gamma-aminobutyric acidergic neurons. Following the immunohistochemical procedures, the mean numbers of labeled cells in the lateral and medial septal nuclei were obtained for the two isoform groups. Statistical analysis yielded significant results which indicated that the 65 isoform of glutamic acid decarboxylase predominates in both lateral and medial septal nuclei (unpaired two-tailed t-test p < 0.0001 for LS, p < 0.01 for MS). This study is the first to reveal the dominance of glutamic acid decarboxylase isoform 65 in the septal region in glutamic acid decarboxylase-green fluorescent protein transgenic mice.

  6. Post-transcriptional regulation of ornithine decarboxylase in Xenopus laevis oocytes.

    PubMed

    Bassez, T; Paris, J; Omilli, F; Dorel, C; Osborne, H B

    1990-11-01

    The level at which ornithine decarboxylase expression is regulated in growing oocytes has been investigated. Immunoprecipitation of the in vivo labelled proteins showed that ornithine decarboxylase accumulated less rapidly in stage IV oocytes than in previtellogenic stage I + II oocytes. Quantitative Northern analysis showed that ornithine decarboxylase mRNA is abundant in oocytes (about 8 x 10(8) transcripts/cell) and this number does not significantly change during oogenesis. Polysome analysis showed that this mRNA is present in polysomes in stage I + II oocytes but has passed into puromycin-insensitive mRNP particles by stage IV of oogenesis. Therefore, during the growth phase of oogenesis, ornithine decarboxylase expression is regulated at a translational level. These results are discussed relative to the temporal expression of ornithine decarboxylase and of other proteins whose expression also decreases during oogenesis. In order to perform these experiments, the cDNA (XLODC1) corresponding to Xenopus laevis ornithine decarboxylase mRNA was cloned and sequenced.

  7. Structure and Function of 4-Hydroxyphenylacetate Decarboxylase and Its Cognate Activating Enzyme.

    PubMed

    Selvaraj, Brinda; Buckel, Wolfgang; Golding, Bernard T; Ullmann, G Matthias; Martins, Berta M

    2016-01-01

    4-Hydroxyphenylacetate decarboxylase (4Hpad) is the prototype of a new class of Fe-S cluster-dependent glycyl radical enzymes (Fe-S GREs) acting on aromatic compounds. The two-enzyme component system comprises a decarboxylase responsible for substrate conversion and a dedicated activating enzyme (4Hpad-AE). The decarboxylase uses a glycyl/thiyl radical dyad to convert 4-hydroxyphenylacetate into p-cresol (4-methylphenol) by a biologically unprecedented Kolbe-type decarboxylation. In addition to the radical dyad prosthetic group, the decarboxylase unit contains two [4Fe-4S] clusters coordinated by an extra small subunit of unknown function. 4Hpad-AE reductively cleaves S-adenosylmethionine (SAM or AdoMet) at a site-differentiated [4Fe-4S]2+/+ cluster (RS cluster) generating a transient 5'-deoxyadenosyl radical that produces a stable glycyl radical in the decarboxylase by the abstraction of a hydrogen atom. 4Hpad-AE binds up to two auxiliary [4Fe-4S] clusters coordinated by a ferredoxin-like insert that is C-terminal to the RS cluster-binding motif. The ferredoxin-like domain with its two auxiliary clusters is not vital for SAM-dependent glycyl radical formation in the decarboxylase, but facilitates a longer lifetime for the radical. This review describes the 4Hpad and cognate AE families and focuses on the recent advances and open questions concerning the structure, function and mechanism of this novel Fe-S-dependent class of GREs.

  8. High levels of acetoacetate and glucose increase expression of cytokines in bovine hepatocytes, through activation of the NF-κB signalling pathway.

    PubMed

    Li, Yu; Ding, Hongyan; Wang, Xichun; Liu, Lei; Huang, Dan; Zhang, Renhe; Guo, Lihui; Wang, Zhe; Li, Xiaobing; Liu, Guowen; Wu, Jinjie; Li, Xinwei

    2016-02-01

    Elevated levels of blood interleukin-1β (IL-1β), interleukin-6 (IL-6) and tumour necrosis factor-α (TNF-α) increase insulin resistance and result in inflammation. It is not clear whether elevated blood level of acetoacetate (ACAC) and decreased blood level of glucose, which are the predominant characteristics of clinical biochemistry in ketotic dairy cows, increase proinflammatory cytokines and subsequent inflammation. The objective of this study was to test the hypothesis that ACAC and glucose activate the NF-κB signalling pathway to regulate cytokines expression in bovine hepatocytes. Bovine hepatocytes were cultured with ACAC (0-4.8 mm) and glucose (0-5.55 mm) with or without NF-κB inhibitor PDTC for 24 h. The secretion and mRNA levels of cytokines were determined by enzyme-linked immunosorbent assay (ELISA) and real-time fluorescence quantitative polymerase chain reaction (qRT-PCR). The NF-κB signalling pathway activation was evaluated by western blotting. Results showed that the secretion and expression of IL-1β, IL-6 and TNF-α increased in an ACAC dose-dependent manner. Additionally, there was an increase in the secretion and mRNA expression of these three cytokines in glucose treatment group, which increased significantly when the glucose concentrations exceed 3.33 mm. Furthermore, both ACAC and glucose upregulated NF-κB p65 protein expression and IκBα phosphorylation levels. However, these effects were reduced by PDTC. These results demonstrate that elevated levels of ACAC and glucose increase the synthesis and expression of proinflammatory factors by activating NF-κB signalling pathway in hepatocytes, which may contribute to inflammation injury in ketotic dairy cows.

  9. Responses of enzymatic antioxidants and non-enzymatic antioxidants in the cyanobacterium Microcystis aeruginosa to the allelochemical ethyl 2-methyl acetoacetate (EMA) isolated from reed (Phragmites communis).

    PubMed

    Hong, Yu; Hu, Hong-Ying; Xie, Xing; Li, Feng-Min

    2008-08-25

    Macrophytic allelochemicals are considered an environment-friendly and promising alternative to control algal bloom. However, studies examining the potential mechanisms of inhibitory allelochemicals on algae are few. The allelochemical ethyl 2-methyl acetoacetate (EMA), isolated from reed (Phragmites communis), was a strong allelopathic inhibitor on the growth of Microcystis aeruginosa. EMA-induced antioxidant responses were investigated in the cyanobacterium M. aeruginosa to understand the mechanism of EMA inhibition on algal growth. The activities of enzymatic antioxidants superoxide dismutase (SOD) and catalase (CAT), and the contents of non-enzymatic antioxidants reduced glutathione (GSH) and ascorbic acid (AsA) of M. aeruginosa cells were analyzed after treatments with different concentrations of EMA. Exposure of M. aeruginosa to EMA caused changes in enzyme activities and contents of non-enzymatic antioxidants in different manners. The decrease in SOD activity occurred first after 4 h of EMA exposure, and more markedly after 40 h. CAT activity did not change after 4 h of EMA exposure, but increased obviously after 40 h. The contents of AsA and GSH were increased greatly by EMA after 4 h. After 60 h, low EMA concentrations still increased the CAT activity and the contents of AsA and GSH, but high EMA concentrations started to impose a marked suppression on them. EMA increased dehydroascorbate (DHAsA) and oxidized glutathione (GSSG) contents during all exposure times. After 60 h, the regeneration rates of AsA and GSH (represented by the AsA/DHAsA ratio and GSH/GSSG ratio, respectively) were reduced by high EMA concentrations. These results suggest that the activation of CAT and the availability of AsA and GSH at early exposure are important to counteract the oxidative stress induced by EMA, and the inactivation of SOD may be crucial to the growth inhibition of M. aeruginosa by EMA.

  10. Neuroprotection by acetoacetate and β-hydroxybutyrate against NMDA-induced RGC damage in rat—possible involvement of kynurenic acid

    PubMed Central

    Thaler, Sebastian; Choragiewicz, Tomasz J.; Rejdak, Robert; Fiedorowicz, Michal; Turski, Waldemar A.; Tulidowicz-Bielak, Maria; Zrenner, Eberhart; Schuettauf, Frank

    2010-01-01

    Purpose This study investigated the effects of systemically administered lithium acetoacetate (ACA) and sodium β-hydroxybutyrate (BHB) in a rat model of N-methyl-D-aspartate (NMDA)-induced damage of retinal ganglion cells (RGC). Additionally, the influence of ACA and BHB on kynurenic acid (KYNA) production was assessed in vitro in bovine retinal slices. Methods Female adult Brown–Norway rats in groups of 5–8 animals were used. ACA and BHB were administered intraperitoneally once a day for 21 consecutive days, and phosphate buffered saline (PBS) was administered to control animals. After 2 weeks, the animals received intraocular NMDA (2 μl of a 10 mM solution in PBS) or intraocular PBS as a control. On day 19, retinal ganglion cells were labeled retrogradely with hydroxystilbamidine. Two days later, RGC density (cells per mm2) was assessed on retinal flatmounts. Additionaly, bovine retinal slices were incubated with NMDA and ACA or BHB at concentrations of 1.0 mM and 3.0 mM, and de novo KYNA production was measured using HPLC. Results Intraperitoneal ACA (250 mg/kg) or BHB (291.2 mg/kg) significantly protected RGC against NMDA-induced neurodegeneration. De novo KYNA production in bovine retinal slices was lowered by NMDA. Both ACA and BHB at a concentration of 3.0 mM significantly reduced the effects of NMDA. Conclusions ACA and BHB had a significant dose-dependent neuroprotective effect on RGC in a rat model of NMDA-induced RGC damage. Both ketone bodies also significantly attenuated NMDA-induced reduction of retinal KYNA production in vitro, suggesting that this mechanism may be essential for the neuroprotective effects of ACA and BHB in vivo. Our results imply that ketone bodies may represent an additional treatment option in chronic neurodegenerative disorders of the eye. PMID:20532550

  11. Effect of Hyperketonemia (Acetoacetate) on Nuclear Factor-κB and p38 Mitogen-Activated Protein Kinase Activation Mediated Intercellular Adhesion Molecule 1 Upregulation in Endothelial Cells

    PubMed Central

    Rains, Justin L.

    2015-01-01

    Abstract Background: Hyperketonemia is a pathological condition observed in patients with type 1 diabetes and ketosis-prone diabetes (KPD), which results in increased blood levels of acetoacetate (AA) and β-hydroxybutyrate (BHB). Frequent episodes of hyperketonemia are associated with a higher incidence of vascular disease. We examined the hypothesis that hyperketonemia activates the nuclear factor-κB (NF-κB) and mitogen-activated protein kinase (MAPK) signaling pathways that regulate intercellular adhesion molecule 1 (ICAM-1) expression in endothelial cells. Methods: Human umbilical vein endothelial cells (HUVECs) were cultured with AA (0–8 mM) or BHB (0–10 mM) for 0–24 hr. Western blotting was used to determine NF-κB activation in whole-cell lysates. ICAM-1 expression was measured using flow cytometry. Results: Results show a 2.4-fold increase in NF-κB activation in cells treated with 8 mM AA compared to the control. BHB had little or no effect on NF-κB activation. Pretreatment with a reactive oxygen species (ROS) inhibitor [N-acetyl-l-cysteine (NAC)] reduced NF-κB to near-control levels. The expression of AA-induced ICAM-1 was significantly reduced when cells were pretreated with either NAC or p38 MAPK inhibitor. Conclusions: These results suggest that NF-κB and p38 MAPK mediate upregulation of ICAM-1 expression in endothelial cells exposed to elevated levels of AA, which may contribute to the development of vascular disease in diabetes. PMID:25489974

  12. Mild experimental ketosis increases brain uptake of 11C-acetoacetate and 18F-fluorodeoxyglucose: a dual-tracer PET imaging study in rats.

    PubMed

    Pifferi, Fabien; Tremblay, Sébastien; Croteau, Etienne; Fortier, Mélanie; Tremblay-Mercier, Jennifer; Lecomte, Roger; Cunnane, Stephen C

    2011-03-01

    Brain glucose and ketone uptake was investigated in Fisher rats subjected to mild experimental ketonemia induced by a ketogenic diet (KD) or by 48 hours fasting (F). Two tracers were used, (11)C-acetoacetate ((11)C-AcAc) for ketones and (18)F-fluorodeoxyglucose for glucose, in a dual-tracer format for each animal. Thus, each animal was its own control, starting first on the normal diet, then undergoing 48 hours F, followed by 2 weeks on the KD. In separate rats on the same diet conditions, expression of the transporters of glucose and ketones (glucose transporter 1 (GLUT1) and monocarboxylic acid transporter (MCT1)) was measured in brain microvessel preparations. Compared to controls, uptake of (11)C-AcAc increased more than 2-fold while on the KD or after 48 hours F (P < 0.05). Similar trends were observed for (18)FDG uptake with a 1.9-2.6 times increase on the KD and F, respectively (P < 0.05). Compared to controls, MCT1 expression increased 2-fold on the KD (P < 0.05) but did not change during F. No significant difference was observed across groups for GLUT1 expression. Significant differences across the three groups were observed for plasma beta-hydroxybutyrate (beta-HB), AcAc, glucose, triglycerides, glycerol, and cholesterol (P < 0.05), but no significant differences were observed for free fatty acids, insulin, or lactate. Although the mechanism by which mild ketonemia increases brain glucose uptake remains unclear, the KD clearly increased both the blood-brain barrier expression of MCT1 and stimulated brain (11)C-AcAc uptake. The present dual-tracer positron emission tomography approach may be particularly interesting in neurodegenerative pathologies such as Alzheimer's disease where brain energy supply appears to decline critically.

  13. High glucose and ketosis (acetoacetate) increases, and chromium niacinate decreases, IL-6, IL-8, and MCP-1 secretion and oxidative stress in U937 monocytes.

    PubMed

    Jain, Sushil K; Rains, Justin L; Croad, Jennifer L

    2007-10-01

    Elevated blood levels of the proinflammatory cytokines interleukin-6 (IL-6), interleukin-8 (IL-8), and MCP-1 (monocyte chemoattractant protein-1) increase insulin resistance and the risk of cardiovascular disease (CVD). There is no previous study that has examined the effect of ketosis and trivalent chromium on IL-6, IL-8, or MCP-1 secretion in any cell type or in human or animal model. The authors examined the hypothesis that ketosis increases and trivalent chromium decreases the levels of cytokines and oxidative stress in diabetes using a U937 monocyte cell culture model. Cells were cultured with control, high glucose (HG), and acetoacetate (AA) in the absence or presence (0.5-10 microM) of CrCl(3), chromium picolinate (Cr-P), or chromium niacinate (Cr-N) at 37 degrees C for 24 h. The data show a significant stimulation of IL-6, IL-8, and MCP-1 secretion and an increase in oxidative stress in cells treated with HG or AA. The effect of HG on cytokine secretion was reduced by Cr-N, and to a lesser extent by CrCl(3) and Cr-P. The effect of HG on oxidative stress was reduced by Cr-N and CrCl 3, but not by Cr-P. Similarly, Cr-N decreased the cytokine secretion in HG + AA-treated cells. Cr-N significantly decreased standard oxidant (H(2)O(2)) induced cytokine secretion, which suggests that reduction of cytokine secretion by Cr-N is in part mediated by its antioxidative effect. In a cell culture model, Cr-N appears to be the most effective form of chromium in inhibiting oxidative stress and proinflammatory cytokine secretion by monocytes. This study suggests that chromium niacinate supplementation may be useful in reducing vascular inflammation and the risk of CVD in diabetes.

  14. MO-F-CAMPUS-I-05: Quantitative ADC Measurement of Esophageal Cancer Before and After Chemoradiation

    SciTech Connect

    Yang, L; Son, JB; Ma, J; Hazle, J; Carter, BW; Lin, S; Cheng, S

    2015-06-15

    Purpose: We investigated whether quantitative diffusion imaging can be used as an imaging biomarker for early prediction of treatment response of esophageal cancer. Methods: Eight patients with esophageal cancer underwent a baseline and an interim MRI studies during chemoradiation on a 3T whole body MRI scanner with an 8-channel torso phased array coil. Each MRI study contained two axial diffusion-weighted imaging (DWI) series with a conventional DWI sequence and a reduced field-of-view DWI sequence (FOCUS) of varying b-values. ADC maps with two b-values were computed from conventional DWI images using a mono-exponential model. For each of DWI sequences, separate ADCall was computed by fitting the signal intensity of images with all the b-values to a single exponential model. For the FOCUS sequence, a bi-exponential model was used to extract perfusion and diffusion coefficients (ADCperf and ADCdiff) and their contributions to the signal decay. A board-certified radiologist contoured the tumor region and mean ADC values and standard deviations of tumor and muscle ROIs were recorded from different ADC maps. Results: Our results showed that (1) the magnitude of ADCs from the same ROIs by the different analysis methods can be substantially different. (2) For a given method, the change between the baseline and interim muscle ADCs was relatively small (≤10%). In contrast, the change between the baseline and interim tumor ADCs was substantially larger, with the change in ADCdiff by FOCUS DWI showing the largest percentage change of 73.2%. (3) The range of the relative change of a specific parameter for different patients was also different. Conclusion: Presently, we do not have the final pathological confirmation of the treatment response for all the patients. However, for a few patients whose surgical specimen is available, the quantitative ADC changes have been found to be useful as a potential predictor for treatment response.

  15. Utility of ADC measurement on diffusion-weighted MRI in differentiation of prostate cancer, normal prostate and prostatitis.

    PubMed

    Esen, Meltem; Onur, Mehmet Ruhi; Akpolat, Nusret; Orhan, Irfan; Kocakoc, Ercan

    2013-08-01

    To determine the utility of apparent diffusion coefficient (ADC) values in differentiation of prostate cancer from normal prostate parenchyma and prostatitis we obtained ADC values of 50 patients at b 100, 600 and 1,000 s/mm(2) diffusion gradients. The ADC values of prostate cancer group were significantly lower than normal prostate and prostatitis group at b 600 and 1,000 s/mm(2) gradients. The ADC values at high diffusion gradients may be used in differentiation prostate cancer from normal prostate and prostatitis.

  16. In-line e-beam inspection with optimized sampling and newly developed ADC

    NASA Astrophysics Data System (ADS)

    Ikota, Masami; Miura, Akihiro; Fukunishi, Munenori; Hiroi, Takashi; Sugimoto, Aritoshi

    2003-07-01

    An electron beam inspection is strongly required for HARI to detect contact and via defects that an optical inspection cannot detect. Conventionally, an e-beam inspection system is used as an analytical tool for checking the process margin. Due to its low throughput speed, it has not been used for in-line QC. Therefore, we optimized the inspection area and developed a new auto defect classification (ADC) to use with e-beam inspection as an in-line inspection tool. A 10% interval scan sampling proved able to estimate defect densities. Inspection could be completed within 1 hour. We specifically adapted the developed ADC for use with e-beam inspection because the voltage contrast images were not sufficiently clear so that classifications could not be made with conventional ADC based on defect geometry. The new ADC used the off-pattern area of the defect to discriminate particles from other voltage contrast defects with an accuracy of greater than 90%. Using sampling optimization and the new ADC, we achieved inspection and auto defect review with throughput of less than 1 and one-half hours. We implemented the system as a procedure for product defect QC and proved its effectiveness for in-line e-beam inspection.

  17. Histogram-based classification with Gaussian mixture modeling for GBM tumor treatment response using ADC map

    NASA Astrophysics Data System (ADS)

    Huo, Jing; Kim, Hyun J.; Pope, Whitney B.; Okada, Kazunori; Alger, Jeffery R.; Wang, Yang; Goldin, Jonathan G.; Brown, Matthew S.

    2009-02-01

    This study applied a Gaussian Mixture Model (GMM) to apparent diffusion coefficient (ADC) histograms to evaluate glioblastoma multiforme (GBM) tumor treatment response using diffusion weighted (DW) MR images. ADC mapping, calculated from DW images, has been shown to reveal changes in the tumor's microenvironment preceding morphologic tumor changes. In this study, we investigated the effectiveness of features that represent changes from pre- and post-treatment tumor ADC histograms to detect treatment response. The main contribution of this work is to model the ADC histogram as the composition of two components, fitted by GMM with expectation maximization (EM) algorithm. For both pre- and post-treatment scans taken 5-7 weeks apart, we obtained the tumor ADC histogram, calculated the two-component features, as well as the other standard histogram-based features, and applied supervised learning for classification. We evaluated our approach with data from 85 patients with GBM under chemotherapy, in which 33 responded and 52 did not respond based on tumor size reduction. We compared AdaBoost and random forests classification algorithms, using ten-fold cross validation, resulting in a best accuracy of 69.41%.

  18. High-Speed, Multi-Channel Serial ADC LVDS Interface for Xilinx Virtex-5 FPGA

    NASA Technical Reports Server (NTRS)

    Taylor, Gregory H.

    2012-01-01

    Analog-to-digital converters (ADCs) are used in scientific and communications instruments on all spacecraft. As data rates get higher, and as the transition is made from parallel ADC designs to high-speed, serial, low-voltage differential signaling (LVDS) designs, the need will arise to interface these in field programmable gate arrays (FPGAs). As Xilinx has released the radiation-hardened version of the Virtex-5, this will likely be used in future missions. High-speed serial ADCs send data at very high rates. A de-serializer instantiated in the fabric of the FPGA could not keep up with these high data rates. The Virtex-5 contains primitives designed specifically for high-speed, source-synchronous de-serialization, but as supported by Xilinx, can only support bitwidths of 10. Supporting bit-widths of 12 or more requires the use of the primitives in an undocumented configuration, a non-trivial task. A new SystemVerilog design was written that is simpler and uses fewer hardware resources than the reference design described in Xilinx Application Note XAPP866. It has been shown to work in a Xilinx XC5VSX24OT connected to a MAXIM MAX1438 12-bit ADC using a 50-MHz sample clock. The design can be replicated in the FPGA for multiple ADCs (four instantiations were used for a total of 28 channels).

  19. Development, integrated investigation, laboratory and in-flight testing of Chibis-M microsatellite ADCS

    NASA Astrophysics Data System (ADS)

    Ovchinnikov, M. Yu.; Ivanov, D. S.; Ivlev, N. A.; Karpenko, S. O.; Roldugin, D. S.; Tkachev, S. S.

    2014-01-01

    Design, analytical investigation, laboratory and in-flight testing of the attitude determination and control system (ADCS) of a microsatellites are considered. The system consists of three pairs of reaction wheels, three magnetorquers, a set of Sun sensors, a three-axis magnetometer and a control unit. The ADCS is designed for a small 10-50 kg LEO satellite. System development is accomplished in several steps: satellite dynamics preliminary study using asymptotical and numerical techniques, hardware and software design, laboratory testing of each actuator and sensor and the whole ADCS. Laboratory verification is carried out on the specially designed test-bench. In-flight ADCS exploitation results onboard the Russian microsatellite "Chibis-M" are presented. The satellite was developed, designed and manufactured by the Institute of Space Research of RAS. "Chibis-M" was launched by the "Progress-13M" cargo vehicle on January 25, 2012 after undocking from the International Space Station (ISS). This paper assess both the satellite and the ADCS mock-up dynamics. Analytical, numerical and laboratory study results are in good correspondence with in-flight data.

  20. Photoluminescence detection of alpha particle using DAM-ADC nuclear detector

    NASA Astrophysics Data System (ADS)

    Abdalla, Ayman M.; Harraz, Farid A.; Ali, Atif M.; Al-Sayari, S. A.; Al-Hajry, A.

    2016-09-01

    The photoluminescence (PL) and UV-vis spectral analysis of DAM-ADC (diallyl maleate: DAM, polyallyl diglycol carbonate: ADC) nuclear detector are demonstrated for the first time. The DAM-ADC surfaces were exposed to thin 241Am disk source that emits alpha particles with activity 333 kBq. It is found that the track density of the irradiated samples remarkably influences the PL characteristics of the DAM-ADC detector. The spectral peak heights and the integrated intensities under the peaks exhibit linear correlations with correlation coefficient R2=0.9636 and 0.9806, respectively for different alpha particle fluences ranging from 8.16-40.82×107 particles/cm2. Additionally, a correlation coefficient R2=0.9734 was achieved for the UV-vis spectral analysis. The linear fitting functions, along with the corresponding fitting parameters were evaluated in each case. Both the PL and the UV-vis data of the irradiated DAM-ADC samples showed considerable spectral differences, and hence they would be used to offer sensitive approaches for alpha particle detection.

  1. A low power 11-bit 100 MS/s SAR ADC IP

    NASA Astrophysics Data System (ADS)

    Ya, Wang; Chunying, Xue; Fule, Li; Chun, Zhang; Zhihua, Wang

    2015-02-01

    This paper presents a dual-channel 11-bit 100 MS/s hybrid SAR ADC IP. Each channel adopts flash-SAR architecture for high speed, low power and high linearity. Dynamic comparators in the coarse flash ADC and the fine SAR ADC further contribute to the reduction of power consumption. A gate-controlled ring oscillator generates a multi-phase clock for SAR logic, thereby allowing it to asynchronously trigger the comparator in the fine SAR ADC in high speed. MOM capacitors with a fully shielded structure provide enough matching accuracy without the need for calibration. This design was fabricated in SMIC 55 nm low leakage CMOS technology and the active area of dual-channel (I-Q) ADC is 0.35 mm2, while the core area is 0.046 mm2. It consumes 2.92 mA at a 1.2 V supply, for dual-channel too. The effective number of bits (ENOB) is 9.90 bits at 2.4 MHz input frequency, and 9.34 bits at 50 MHz, leading to a FOM of 18.3 fJ/conversion-step.

  2. A 10-bit 50-MS/s subsampling pipelined ADC based on SMDAC and opamp sharing

    NASA Astrophysics Data System (ADS)

    Lijie, Chen; Yumei, Zhou; Baoyue, Wei

    2010-11-01

    This paper describes a 10-bit, 50-MS/s pipelined A/D converter (ADC) with proposed area- and power-efficient architecture. The conventional dedicated sample-hold-amplifier (SHA) is eliminated and the matching requirement between the first multiplying digital-to-analog converter (MDAC) and sub-ADC is also avoided by using the SHA merged with the first MDAC (SMDAC) architecture, which features low power and stabilization. Further reduction of power and area is achieved by sharing an opamp between two successive pipelined stages, in which the effect of opamp offset and crosstalk between stages is decreased. So the 10-bit pipelined ADC is realized using just four opamps. The ADC demonstrates a maximum signal-to-noise distortion ratio and spurious free dynamic range of 52.67 dB and 59.44 dB, respectively, with a Nyquist input at full sampling rate. Constant dynamic performance for input frequencies up to 49.7 MHz, which is the twofold Nyquist rate, is achieved at 50 MS/s. The ADC prototype only occupies an active area of 1.81 mm2 in a 0.35 μm CMOS process, and consumes 133 mW when sampling at 50 MHz from a 3.3-V power supply.

  3. Fast detection of alpha particles in DAM-ADC nuclear track detectors

    NASA Astrophysics Data System (ADS)

    Rammah, Y. S.; Ashraf, O.; Abdalla, A. M.; Eisa, M.; Ashry, A. H.; Tsuruta, T.

    2015-02-01

    Fast detection of alpha particles in DAM-ADC nuclear track detectors using a new chemical etchant was investigated. 252Cf and 241Am sources were used for irradiating samples of DAM-ADC SSNTDs with fission fragments and alpha particles in air at normal temperature and pressure. A series of experimental chemical etching are carried out using new etching solution (8 ml of 10 N NaOH+ 1 ml CH3OH) at 60 °C to detect alpha particle in short time in DAM-ADC detectors. Suitable analyzing software has been used to analyze experimental data. From fission and alpha track diameters, the value of bulk etching rate is equal to 8.52 μm/h. Both of the sensitivity and etching efficiency were found to vary with the amount of methanol in the etching solution and etching time. The DAM-ADC detectors represent the best efficiency applicable in detectors in the entire range of alpha energies (from 1 to 5 MeV). The activation energies of this etchant have been calculated; track activation energy, ET, has been found to be lower than the bulk activation energy, EB, for the DAM-ADC nuclear track detectors. These results are in more agreement with the previous work.

  4. Anti-glutamic acid decarboxylase antibody positive neurological syndromes

    PubMed Central

    Tohid, Hassaan

    2016-01-01

    A rare kind of antibody, known as anti-glutamic acid decarboxylase (GAD) autoantibody, is found in some patients. The antibody works against the GAD enzyme, which is essential in the formation of gamma aminobutyric acid (GABA), an inhibitory neurotransmitter found in the brain. Patients found with this antibody present with motor and cognitive problems due to low levels or lack of GABA, because in the absence or low levels of GABA patients exhibit motor and cognitive symptoms. The anti-GAD antibody is found in some neurological syndromes, including stiff-person syndrome, paraneoplastic stiff-person syndrome, Miller Fisher syndrome (MFS), limbic encephalopathy, cerebellar ataxia, eye movement disorders, and epilepsy. Previously, excluding MFS, these conditions were called ‘hyperexcitability disorders’. However, collectively, these syndromes should be known as “anti-GAD positive neurological syndromes.” An important limitation of this study is that the literature is lacking on the subject, and why patients with the above mentioned neurological problems present with different symptoms has not been studied in detail. Therefore, it is recommended that more research is conducted on this subject to obtain a better and deeper understanding of these anti-GAD antibody induced neurological syndromes. PMID:27356651

  5. Multiple roles of the active site lysine of Dopa decarboxylase.

    PubMed

    Bertoldi, Mariarita; Voltattorni, Carla Borri

    2009-08-15

    The pyridoxal 5'-phosphate dependent-enzyme Dopa decarboxylase, responsible for the irreversible conversion of l-Dopa to dopamine, is an attractive drug target. The contribution of the pyridoxal-Lys303 to the catalytic mechanisms of decarboxylation and oxidative deamination is analyzed. The K303A variant binds the coenzyme with a 100-fold decreased apparent equilibrium binding affinity with respect to the wild-type enzyme. Unlike the wild-type, K303A in the presence of l-Dopa displays a parallel progress course of formation of both dopamine and 3,4-dihydroxyphenylacetaldehyde (plus ammonia) with a burst followed by a linear phase. Moreover, the finding that the catalytic efficiencies of decarboxylation and of oxidative deamination display a decrease of 1500- and 17-fold, respectively, with respect to the wild-type, is indicative of a different impact of Lys303 mutation on these reactions. Kinetic analyses reveal that Lys303 is involved in external aldimine formation and hydrolysis as well as in product release which affects the rate-determining step of decarboxylation.

  6. Studies on uroporphyrinogen decarboxylase from Chlorella kessleri (Trebouxiophyceae, Chlorophyta).

    PubMed

    Juárez, Angela B; Aldonatti, Carmen; Vigna, María S; Ríos de Molina, María Del C

    2007-02-01

    Uroporphyrinogen decarboxylase (UroD) (EC 4.1.1.37) is an enzyme from the tetrapyrrole biosynthetic pathway, in which chlorophyll is the main final product in algae. This is the first time that a study on UroD activity has been performed in a green alga (Chlorella). We isolated and partially purified the enzyme from a Chlorella kessleri (Trebouxiophyceae, Chlorophyta) strain (Copahue, Neuquén, Argentina), and describe for the first time some of its properties. In C. kessleri, the decarboxylation of uroporphyrinogen III occurs in two stages, via 7 COOH and then 6 and 5 COOH intermediates, with the decarboxylation of the 7 COOH compound being the rate-limiting step for the reaction. Cultures in the exponential growth phase showed the highest specific activity values. The most suitable conditions to measure UroD activity in C. kessleri were as follows: 0.23-0.3 mg protein/mL, approximately 6-8 micromol/L uroporphyrinogen III, and 20 min incubation time. Gel filtration chromatography and Western blot assays indicated that UroD from C. kessleri is a dimer of approximately 90 kDa formed by species of lower molecular mass, which conserves enzymatic activity.

  7. Chemical modification of oxalate decarboxylase to improve adsorption capacity.

    PubMed

    Lin, Rihui; He, Junbin; Wu, Jia; Cai, Xinghua; Long, Han; Chen, Shengfeng; Liu, Haiqian

    2017-02-03

    In order to enhance the adsorption capacity of oxalate decarboxylase (Oxdc) on calcium oxalate monohydrate crystals and improve the application performance of Oxdc, chemical modification of Oxdc with ethylenediaminetetraacetic dianhydride (EDTAD) was investigated in this work. The sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and liquid chromatography tandem mass spectrometry (LC/MS) analysis results demonstrated that Oxdc and EDTAD have been covalently bound, and suggested that the chemical modification occurred at the free amino of the side chain and the α-amine of the N-terminus of Oxdc. Fluorescene and circular dichroic measurement showed that the structure and conformation of Oxdc were tinily altered after modification by EDTAD. The optimum pH of EDTAD-modified Oxdc was shifted to the alkaline side about 1.5 unit and it has a higher thermostability. The analysis of kinetic parameters indicated that the EDTAD-modified Oxdc showed a higher affinity towards the substrate. Through modification the adsorption capacity of Oxdc onto CaOx monohydrate crystals was increased by 42.42%.

  8. Glutamate decarboxylase from Lactobacillus brevis: activation by ammonium sulfate.

    PubMed

    Hiraga, Kazumi; Ueno, Yoshie; Oda, Kohei

    2008-05-01

    In this study, the glutamate decarboxylase (GAD) gene from Lactobacillus brevis IFO12005 (Biosci. Biotechnol. Biochem., 61, 1168-1171 (1997)), was cloned and expressed. The deduced amino acid sequence showed 99.6% and 53.1% identity with GAD of L. brevis ATCC367 and L. lactis respectively. The His-tagged recombinant GAD showed an optimum pH of 4.5-5.0, and 54 kDa on SDS-PAGE. The GAD activity and stability was significantly dependent on the ammonium sulfate concentration, as observed in authentic GAD. Gel filtration showed that the inactive form of the GAD was a dimer. In contrast, the ammonium sulfate-activated form was a tetramer. CD spectral analyses at pH 5.5 revealed that the structures of the tetramer and the dimer were similar. Treatment of the GAD with high concentrations of ammonium sulfate and subsequent dilution with sodium glutamate was essential for tetramer formation and its activation. Thus the biochemical properties of the GAD from L. brevis IFO12005 were significantly different from those from other sources.

  9. Molecular cloning and expression of the mouse ornithine decarboxylase gene.

    PubMed Central

    McConlogue, L; Gupta, M; Wu, L; Coffino, P

    1984-01-01

    We used mRNA from a mutant S49 mouse lymphoma cell line that produces ornithine decarboxylase (OrnDCase) as its major protein product to synthesize and clone cDNA. Plasmids containing OrnDCase cDNA were identified by hybrid selection of OrnDCase mRNA and in vitro translation. The two of these with the largest inserts together span 2.05 kilobases of cDNA. Southern blot analysis of DNA from wild-type or mutant S49 cells, cleaved with EcoRI or with BamHI, revealed multiple bands homologous to OrnD-Case cDNA, only one of which was amplified in the mutant cells. RNA transfer blot analysis showed that the major OrnD-Case mRNA in the mouse lymphoma cells is 2.0 kilobases long. A similar size mRNA was found in mouse kidney and was more abundant in the kidneys of mice treated with testosterone, an inducer of OrnDCase activity in that tissue. Images PMID:6582509

  10. Expression of ornithine decarboxylase in precancerous and cancerous gastric lesions

    PubMed Central

    Miao, Xin-Pu; Li, Jian-Sheng; Li, Hui-Yan; Zeng, Shi-Ping; Zhao, Ye; Zeng, Jiang-Zheng

    2007-01-01

    AIM: To investigate the expression of ornithine decarboxylase (ODC) in precancerous and cancerous gastric lesions. METHODS: We studied the expression of ODC in gastric mucosa from patients with chronic superficial gastritis (CSG, n = 32), chronic atrophic gastritis [CAG, n = 43; 15 with and 28 without intestinal metaplasia (IM)], gastric dysplasia (DYS, n = 11) and gastric cancer (GC, n = 48) tissues using immunohistochemical staining. All 134 biopsy specimens of gastric mucosa were collected by gastroscopy. METHODS: The positive rate of ODC expression was 34.4%, 42.9%, 73.3%, 81.8% and 91.7% in cases with CSG, CAG without IM, CAG with IM, DYS and GC, respectively (P < 0.01), The positive rate of ODC expression increased in the order of CSG < CAG (without IM) < CAG (with IM) < DYS and finally, GC. In addition, ODC positive immunostaining rate was lower in well-differentiated GC than in poorly-differentiated GC (P < 0.05). CONCLUSION: The expression of ODC is positively correlated with the degree of malignity of gastric mucosa and development of gastric lesions. This finding indicates that ODC may be used as a good biomarker in the screening and diagnosis of precancerous lesions. PMID:17569126

  11. Ornithine decarboxylase as a marker for premalignancy in the stomach.

    PubMed Central

    Patchett, S E; Alstead, E M; Butruk, L; Przytulski, K; Farthing, M J

    1995-01-01

    Assessment of mucosal ornithine decarboxylase (ODC) activity in the human large bowel may be of value as a marker of potential malignant risk. Its value as a marker of premalignancy in the upper gastrointestinal tract is less clear. Using a [14C]-ornithine bioassay, gastric mucosal ODC activity was measured in 32 normal subjects and 22 patients with confirmed gastric cancer. These results were compared with 47 patients at increased risk of upper gastrointestinal malignancy, (32 patients with partial gastric resection, 15 patients with familial adenomatous polyposis). Median ODC activity in normal subjects was 371 pmol/mg protein/h, (interquartile range (IQR), 230-617). There was no variation with age or sex and no relation to Helicobacter pylori status. Normal subjects had significantly lower ODC activity than patients with a gastric resection or confirmed gastric cancer, but similar to patients with familial adenomatous polyposis. Furthermore, no difference in activity was identified between patients with a gastric resection and established gastric cancer. ODC activity was, however, significantly increased in areas of gastric atrophy or intestinal metaplasia, regardless of the clinical group from which the samples were obtained. It is concluded that measurement of mucosal ODC activity does not provide additional predictive information of malignant risk in the stomach and investigation of other potential biomarkers of malignancy is warranted. PMID:7672662

  12. Histidine Decarboxylase Deficiency Prevents Autoimmune Diabetes in NOD Mice

    PubMed Central

    Alkan, Manal; Machavoine, François; Rignault, Rachel; Dam, Julie; Dy, Michel; Thieblemont, Nathalie

    2015-01-01

    Recent evidence has highlighted the role of histamine in inflammation. Since this monoamine has also been strongly implicated in the pathogenesis of type-1 diabetes, we assessed its effect in the nonobese diabetic (NOD) mouse model. To this end, we used mice (inactivated) knocked out for the gene encoding histidine decarboxylase, the unique histamine-forming enzyme, backcrossed on a NOD genetic background. We found that the lack of endogenous histamine in NOD HDC−/− mice decreased the incidence of diabetes in relation to their wild-type counterpart. Whereas the proportion of regulatory T and myeloid-derived suppressive cells was similar in both strains, histamine deficiency was associated with increased levels of immature macrophages, as compared with wild-type NOD mice. Concerning the cytokine pattern, we found a decrease in circulating IL-12 and IFN-γ in HDC−/− mice, while IL-6 or leptin remained unchanged, suggesting that histamine primarily modulates the inflammatory environment. Paradoxically, exogenous histamine given to NOD HDC−/− mice provided also protection against T1D. Our study supports the notion that histamine is involved in the pathogenesis of diabetes, thus providing additional evidence for its role in the regulation of the immune response. PMID:26090474

  13. Ornithine decarboxylase antizyme inhibitor 2 regulates intracellular vesicle trafficking

    SciTech Connect

    Kanerva, Kristiina; Maekitie, Laura T.; Baeck, Nils; Andersson, Leif C.

    2010-07-01

    Antizyme inhibitor 1 (AZIN1) and 2 (AZIN2) are proteins that activate ornithine decarboxylase (ODC), the key enzyme of polyamine biosynthesis. Both AZINs release ODC from its inactive complex with antizyme (AZ), leading to formation of the catalytically active ODC. The ubiquitously expressed AZIN1 is involved in cell proliferation and transformation whereas the role of the recently found AZIN2 in cellular functions is unknown. Here we report the intracellular localization of AZIN2 and present novel evidence indicating that it acts as a regulator of vesicle trafficking. We used immunostaining to demonstrate that both endogenous and FLAG-tagged AZIN2 localize to post-Golgi vesicles of the secretory pathway. Immuno-electron microscopy revealed that the vesicles associate mainly with the trans-Golgi network (TGN). RNAi-mediated knockdown of AZIN2 or depletion of cellular polyamines caused selective fragmentation of the TGN and retarded the exocytotic release of vesicular stomatitis virus glycoprotein. Exogenous addition of polyamines normalized the morphological changes and reversed the inhibition of protein secretion. Our findings demonstrate that AZIN2 regulates the transport of secretory vesicles by locally activating ODC and polyamine biosynthesis.

  14. Interaction of NAP-22 with brain glutamic acid decarboxylase (GAD).

    PubMed

    Maekawa, Shohei; Kobayashi, Yuumi; Odagaki, Sin-Ichi; Makino, Midori; Kumanogoh, Haruko; Nakamura, Shun; Morita, Mitsuhiro; Hayashi, Fumio

    2013-03-14

    NAP-22 (also called BASP1 or CAP-23) is a neuron-enriched protein localized mainly in the synaptic vesicles and the synaptic plasma membrane. Biochemically, it is recovered in the lipid raft fraction. In order to understand the physiological function of the neuronal lipid raft, NAP-22 binding proteins were screened with a pull-down assay. Glutamic acid decarboxylase (GAD) was detected through LC-MS/MS, and Western blotting using a specific antibody confirmed the result. Two isoforms of GAD, GAD65 and GAD67, were expressed in bacteria as GST-fusion forms and the interaction with NAP-22 was confirmed in vitro. Partial co-localization of NAP-22 with GAD65 and GAD67 was also observed in cultured neurons. The binding showed no effect on the enzymatic activity of GAD65 and GAD67. These results hence suggest that NAP-22 could participate in the transport of GAD65 and GAD67 to the presynaptic termini and their retention on the synaptic vesicles as an anchoring protein.

  15. Reactivation of substrate-inactivated brain glutamate decarboxylase.

    PubMed

    Meeley, M P; Martin, D L

    1983-03-01

    The effects of ATP and inorganic phosphate (Pi) on the reactivation of glutamate apodecarboxylase by its cofactor pyridoxal-5'-phosphate (pyridoxal-P) was studied. Apoenzyme was prepared by preincubation with glutamate. Apoenzyme prepared with glutamate alone was reactivated slowly and incompletely by adding a saturating concentration of pyridoxal-P (20 microM). Reactivation was slightly enhanced by 1-10 mM Pi. Reactivation by pyridoxal-P plus Pi was greatly enhanced by the presence of low concentrations (less than 100 microM) of ATP during the preparation of apoenzyme with glutamate. Reactivation was much lower if Pi was omitted. Enhancement of reactivation by ATP was due to its effect during apoenzyme formation, since ATP did not enhance reactivation if added only during reactivation and since the enhancing effect persisted after the removal of free ATP by chromatography on Sephadex G-25 after apoenzyme preparation and before reactivation. Reactivation was inhibited by high concentrations of ATP (greater than 100 microM), possibly by competition of ATP for the cofactor binding site. Four factors (glutamate, pyridoxal-P, ATP, and Pi) control a cycle of inactivation and reactivation that appears to be important in the regulation of brain glutamate decarboxylase.

  16. Arginase, Arginine Decarboxylase, Ornithine Decarboxylase, and Polyamines in Tomato Ovaries (Changes in Unpollinated Ovaries and Parthenocarpic Fruits Induced by Auxin or Gibberellin).

    PubMed Central

    Alabadi, D.; Aguero, M. S.; Perez-Amador, M. A.; Carbonell, J.

    1996-01-01

    Arginase (EC 3.5.3.1) activity has been found in the ovaries and Young fruits of tomato (Lycopersicon esculentum Mill. cv Rutgers).Changes in arginase, arginine decarboxylase (EC 4.1.1.19), and ornithine decarboxylase activity (EC 4.1.1.17) and levels of free and conjugated putrescine, spermidine, and spermine were determined in unpollinated ovaries and in parthenocarpic fruits during the early stages of development induced by 2,4-dichlorophenoxyacetic acid (2,4-D) or gibberellic acid (GA3). Levels of arginase, free spermine, and conjugates of the three polyamines were constant in unpollinated ovaries and characteristic of a presenescent step. A marked decrease in arginase activity, free spermine, and polyamine conjugates was associated with the initiation of fruit growth due to cell division, and when cell expansion was initiated, the absence of arginase indicated a redirection of nitrogen metabolism to the synthesis of arginine. A transient increase in arginine decarboxylase and ornithine decarboxylase was also observed in 2,4-D-induced fruits. In general, 2,4-D treatments produced faster changes than GA3, and without treatment, unpollinated ovaries developed only slightly and senescence was hardly visible. Sensitivity to 2,4-D and GA3 treatment remained for at least 2 weeks postanthesis. PMID:12226441

  17. An 8-Bit 600-MSps Flash ADC Using Interpolating and Background Self-Calibrating Techniques

    NASA Astrophysics Data System (ADS)

    Paik, Daehwa; Asada, Yusuke; Miyahara, Masaya; Matsuzawa, Akira

    This paper describes a flash ADC using interpolation (IP) and cyclic background self-calibrating techniques. The proposed IP technique that is cascade of capacitor IP and gate IP with dynamic double-tail latched comparator reduces non-linearity, power consumption, and occupied area. The cyclic background self-calibrating technique periodically suppresses offset mismatch voltages caused by static fluctuation and dynamic fluctuation due to temperature and supply voltage changes. The ADC has been fabricated in 90-nm 1P10M CMOS technology. Experimental results show that the ADC achieves SNDR of 6.07bits without calibration and 6.74bits with calibration up to 500MHz input signal at sampling rate of 600MSps. It dissipates 98.5mW on 1.2-V supply. FoM is 1.54pJ/conv.

  18. SEMICONDUCTOR INTEGRATED CIRCUITS: An undersampling 14-bit cyclic ADC with over 100-dB SFDR

    NASA Astrophysics Data System (ADS)

    Weitao, Li; Fule, Li; Dandan, Guo; Chun, Zhang; Zhihua, Wang

    2010-02-01

    A high linearity, undersampling 14-bit 357 kSps cyclic analog-to-digital convert (ADC) is designed for a radio frequency identification transceiver system. The passive capacitor error-average (PCEA) technique is adopted for high accuracy. An improved PCEA sampling network, capable of eliminating the crosstalk path of two pipelined stages, is employed. Opamp sharing and the removal of the front-end sample and hold amplifier are utilized for low power dissipation and small chip area. An additional digital calibration block is added to compensate for the error due to defective layout design. The presented ADC is fabricated in a 180 nm CMOS process, occupying 0.65 × 1.6 mm2. The input of the undersampling ADC achieves 15.5 MHz with more than 90 dB spurious free dynamic range (SFDR), and the peak SFDR is as high as 106.4 dB with 2.431 MHz input.

  19. Fast neutrons detection in CR-39 and DAM-ADC nuclear track detectors

    NASA Astrophysics Data System (ADS)

    Abdalla, A. M.; Ashraf, O.; Rammah, Y. S.; Ashry, A. H.; Eisa, M.; Tsuruta, T.

    2015-03-01

    Fast detection of neutrons in CR-39 and DAM-ADC nuclear track detectors were investigated using new etching conditions. The neutron irradiation is performed using a 5 mCi Am-Be source present at the National Institute of Standards (NIS) of Egypt. Using the new etching condition, irradiated CR-39 samples were etched for 4 h and DAM-ADC samples for 80 min. Suitable analyzing software has been used to analyze experimental data.The dependence of neutrons track density on the neutrons fluence is investigated. When etched under optimum conditions, the relationship between track density and fluence is determined which is found to be linear. Detection efficiency has been represented for both SSNTDs and found to be constant with fluence, which reflects the importance of using CR-39 and DAM-ADC detectors in the field of neutron dosimetry. Linear relationship between track density and effective dose is determined.

  20. Etching characteristic studies for the detection of alpha particles in DAM-ADC nuclear track detector

    NASA Astrophysics Data System (ADS)

    El-Samman, H.; Ashry, A. H.; Arafa, W.; Abou-leila, M.; Abdalla, A. M.; Tsuruta, T.

    2014-09-01

    This study reports the characteristic studies for the detection of alpha particles in DAM-ADC nuclear track detector. Several important parameters that control the track formation such as, the bulk etch rate (VB), track etching rate (VT), dependence of VB and VT on etching concentration and temperature have been extensively studied. The activation energy (Eb) of the bulk etching rate for the DAM-ADC sheets has been calculated, the dependence of etching efficiency and sensitivity upon etchant concentrations and temperature has been investigated, registration efficiency of DAM-ADC detector etched at the optimum etching condition has been examined. The detailed studied results presented in this study provide various useful information about the mechanism of track formation in polymers.

  1. A 16-bit sigma-delta ADC applied in micro-machined inertial sensor

    NASA Astrophysics Data System (ADS)

    Qiang, Li; Xiaowei, Liu

    2015-04-01

    This paper presents a low-distortion sigma-delta (Σ-Δ) ADC for micro-machined inertial sensors. The design adopts a single-loop, fourth-order low-pass single-bit modulator with feedforward paths which can ensure the signal transfer lossless and reduce the nonlinearity and power consumption. The chip is manufactured in standard 0.5µm CMOS process, and the area is 2.2mm2. The ADC achieves 108dB signal to noise ratio (SNR) and 110dB dynamic range (DR). Total power consumption is less than 15mW with 5V supply.

  2. On Measurement Uncertainty of ADC Nonlinearities in Oscillation-Based Test

    NASA Astrophysics Data System (ADS)

    Mrak, Peter; Biasizzo, Anton; Novak, Franc

    2011-11-01

    Oscillation-based test (OBT) is one of the approaches for measuring static ADC parameters such as differential nonlinearity (DNL) and integral nonlinearity (INL) that can be implemented in a built-in self-test arrangement. When applying the OBT approach in practice we noticed an inherent measurement uncertainty related to the slope of the ADC input signal in OBT test mode. Experimental environment in Matlab has been set up to study the phenomenon. Experiments with varying values of slope were performed to demonstrate the margins of DNL measurement uncertainty.

  3. Front end processing for a 100 MHZ Flash-ADC-System

    SciTech Connect

    Eckerlin, G.; Elsen, E.; Schmitt, H.V.D.; Wagner, A.; Walter, P.V.

    1986-06-01

    An intelligent interface for readout of a high speed (100 MHz), multichannel Flash-ADC System is described. 3072 FADC channels are controlled and read by a system of 34 microprocessors M68000 placed at two different hierarchical levels. In addition to the readout itself, the processors perform a detailed pulse shape analysis necessary for a compact and manageable data format. The purpose of the system is to exploit the good double tract separation and time resolution provided by Flash-ADCs in conjunction with large drift chamber detectors such as JADE at PETRA and OPAL at LEP. Details of the system presently being installed at JADE are reviewed.

  4. Nucleotide sequence and expression of the Enterobacter aerogenes alpha-acetolactate decarboxylase gene in brewer's yeast.

    PubMed Central

    Sone, H; Fujii, T; Kondo, K; Shimizu, F; Tanaka, J; Inoue, T

    1988-01-01

    The nucleotide sequence of a 1.4-kilobase DNA fragment containing the alpha-acetolactate decarboxylase gene of Enterobacter aerogenes was determined. The sequence contains an entire protein-coding region of 780 nucleotides which encodes an alpha-acetolactate decarboxylase of 260 amino acids. The DNA sequence coding for alpha-acetolactate decarboxylase was placed under the control of the alcohol dehydrogenase I promoter of the yeast Saccharomyces cerevisiae in a plasmid capable of autonomous replication in both S. cerevisiae and Escherichia coli. Brewer's yeast cells transformed by this plasmid showed alpha-acetolactate decarboxylase activity and were used in laboratory-scale fermentation experiments. These experiments revealed that the diacetyl concentration in wort fermented by the plasmid-containing yeast strain was significantly lower than that in wort fermented by the parental strain. These results indicated that the alpha-acetolactate decarboxylase activity produced by brewer's yeast cells degraded alpha-acetolactate and that this degradation caused a decrease in diacetyl production. PMID:3278689

  5. Hepatic ornithine decarboxylase induction by potato glycoalkaloids in rats.

    PubMed

    Caldwell, K A; Grosjean, O K; Henika, P R; Friedman, M

    1991-08-01

    The induction of hepatic ornithine decarboxylase (ODC) activity in rat livers by the potato glycoalkaloids alpha-solanine, alpha-chaconine, and their aglycone solanidine, has been studied. Ip administration of alpha-solanine at 7.5, 15 and 30 mg/kg body weight produced markedly elevated enzyme activity at 4 hr after treatment, with a linear dose response. The increase was four-fold at the lowest dose administered to 12-fold at the highest. ODC activity was measured at 1, 2, 3, 4, 5, 6, 8, and 24hr after alpha-solanine was given. A statistically significant increase in enzyme activity was evident at 3 hr after treatment; maximal activity occurred at 5 hr and was approximately 12 times greater than the dimethylsulphoxide (DMSO) control level. Elevated activities persisted for several hours, decreasing to about one-third of the maximal level at 8 hr. The relative effects of alpha-solanine, alpha-chaconine and solanidine on ODC activities were studied at 4 hr using an equimolar dose of 17 mM/kg body weight. ODC activity induced by alpha-chaconine was higher than that induced by alpha-solanine; the latter activity was two-thirds that of the former. The aglycone solanidine did not induce any increase in activity compared with the DMSO control. ODC activity with dexamethasone, a glucocorticoid, at 4 mg/kg body weight, followed a pattern similar to that of alpha-solanine. However, maximal activity occurred slightly earlier at 4 hr after treatment. The results show that the extent of induced ODC activity depends on the structure of the potato alkaloid.

  6. Bacopa monniera recombinant mevalonate diphosphate decarboxylase: Biochemical characterization.

    PubMed

    Abbassi, Shakeel J; Vishwakarma, Rishi K; Patel, Parth; Kumari, Uma; Khan, Bashir M

    2015-08-01

    Mevalonate diphosphate decarboxylase (MDD; EC 4.1.1.33) is an important enzyme in the mevalonic acid pathway catalyzing the Mg(2+)-ATP dependant decarboxylation of mevalonate 5-diphosphate (MVAPP) to isopentenyl diphosphate (IPP). Bacopa monniera recombinant MDD (BmMDD) protein was overexpressed in Escherichia coli BL21 (DE3) strain and purified to apparent homogeneity. Km and Vmax for MVAPP were 144 μM and 52 U mg(-1) respectively. The values of turnover (kcat) and kcat/Km for mevalonate 5-diphosphate were determined to be 40s(-1) and 2.77×10(5) M(-1) s(-1) and kcat and kcat/Km values for ATP were found to be 30 s(-1) and 2.20×10(4) M(-1) s(-1), respectively. pH activity profile indicated the involvement of carboxylate ion, lysine and arginine for the activity of enzyme. The apparent activation energy for the BmMDD catalyzed reaction was 12.7 kJ mol(-1). Optimum pH and temperature for the forward reaction was found to be 8.0 and 45 °C. The enzyme was most stable at pH 7 at 20 °C with the deactivation rate constant (Kd(*)) of 1.69×10(-4) and half life (t1/2) of 68 h. The cation studies suggested that BmMDD is a cation dependant enzyme and optimum activity was achieved in the presence of Mg(2+).

  7. Conformational stabilization of rat s-adenosylmethionine decarboxylase by putrescine.

    PubMed

    Wada, Makiko; Shirahata, Akira

    2010-01-01

    The activity and processing of mammalian S-adenosylmethionine decarboxylase (AdoMetDC) is stimulated by putrescine. To obtain new insights into the mechanism through which putrescine stimulates AdoMetDC, we investigated conformational changes in rat prostate AdoMetDC in the presence or absence of putrescine. We examined the reactivity of purified rat prostate AdoMetDC to the SH-reagent iodoacetic acid (IAA) and its susceptibility to proteolysis in the presence or absence of putrescine using matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS). The activity of AdoMetDC treated with IAA in the absence of putrescine was reduced, but about 80% of its activity remained after treatment with IAA in the presence of putrescine. In the presence of putrescine, IAA incorporation was 1.9 mol IAA/mol of AdoMetDC α-subunit, while there was no incorporation of IAA in the β-subunit of AdoMetDC. In the absence of putrescine, 5.0 mol of IAA/mol of α-subunit and 0.9 mol of IAA/mol of β-subunit were incorporated. Only Cys292 and Cys310 were carboxymethylated by IAA in the presence of putrescine. In contrast, in the absence of putrescine all cysteines were carboxymethylated by IAA. In addition, putrescine slowed the rate of AdoMetDC degradation by trypsin. These results demonstrate that the conformation of AdoMetDC purified from rat prostate is stabilized by putrescine.

  8. Purification and properties of diaminopimelate decarboxylase from Escherichia coli

    PubMed Central

    White, P. J.; Kelly, Bridget

    1965-01-01

    1. Diaminopimelate decarboxylase from a soluble extract of Escherichia coli A.T.C.C. 9637 was purified 200-fold by precipitation of nucleic acids, fractionation with acetone and then with ammonium sulphate, adsorption on calcium phosphate gel and chromatography on DEAE-cellulose or DEAE-Sephadex. 2. The purified enzyme showed only one component in the ultracentrifuge, with a sedimentation coefficient of 5·4s. One major peak and three much smaller peaks were observed on electrophoresis of the enzyme at pH8·9. 3. The mol.wt. of the enzyme was approx. 200000. The catalytic constant was 2000mol. of meso-diaminopimelic acid decomposed/min./mol. of enzyme, at 37°. The relative rates of decarboxylation at 25°, 37° and 45° were 0·17:1·0:1·6. At 37° the Michaelis constant was 1·7mm and the optimum pH was 6·7–6·8. 4. There was an excess of acidic amino acids over basic amino acids in the enzyme, which was bound only on basic cellulose derivatives at pH6·8. 5. The enzyme had an absolute requirement for pyridoxal phosphate as a cofactor; no other derivative of pyridoxine had activity. A thiol compound (of which 2,3-dimercaptopropan-1-ol was the most effective) was also needed as an activator. 6. In the presence of 2,3-dimercaptopropan-1-ol (1mm), heavy-metal ions (Cu2+, Hg2+) did not inhibit the enzyme, but there was inhibition by several amino acids with analogous structures to diaminopimelate, generally at high concentrations relative to the substrate. Penicillamine was inhibitory at relatively low concentrations; its action was prevented by pyridoxal phosphate. PMID:14343156

  9. A 9 b/12 b 50 MS/s experimental ADC with continuous approximation architecture in 65 nm CMOS

    NASA Astrophysics Data System (ADS)

    Xiaofeng, Guo; Fan, Ye; Junyan, Ren

    2016-10-01

    A 9 bits 50 MS/s 0.5 mW continuous approximation mixed successive approximation (CAR&SAR) ADC is presented. A 12 bits 50 MS/s 0.6 mW CAR&CAR ADC is presented. In the field of low power and high performance ADC, CAR is a new architecture different from SAR. It is faster and easier to get high accuracy. Here we will introduce CAR and its circuit implementation, and the 9 bits experimental ADC is designed to verify CARADC's feasibility. Meanwhile, its resolution can be extended to 12 bits with adding an extra CAR, and then the performance is raised to 0.6 mW 50 MS/s 72 dB SNDR at TT corner and the Walden FOM is 3.6 fj/conv-step. The 9 b ADC was fabricated by using TSMC 1P9M 65 nm CMOS technology. The ADC achieves 50 dB SNDR and the realized Walden FOM is 34 fj/conv-step. The simulation and measurement results prove that CAR is available in the low power and high performance ADC and it even outperforms SAR. The ADC core occupies an active area of 0.045 mm2.

  10. Prognostic value of the primary lesion apparent diffusion coefficient (ADC) in nasopharyngeal carcinoma: a retrospective study of 541 cases

    PubMed Central

    Zhang, Yuan; Liu, Xu; Zhang, Yun; Li, Wen-Fei; Chen, Lei; Mao, Yan-Ping; Shen, Jing-Xian; Zhang, Fan; Peng, Hao; Liu, Qing; Sun, Ying; Ma, Jun

    2015-01-01

    The prognostic value of the primary lesion pretreatment apparent diffusion coefficient (ADC), which is obtained by diffusion-weighted magnetic resonance imaging (MR-DWI), remains unknown in nasopharyngeal carcinoma (NPC). Thus, to investigate whether the pretreatment ADC value as measured from the primary site on MR-DWI is an independent prognostic factor in NPC, we retrospectively reviewed a cohort of 541 patients with histologically-proven stage I-IVB NPC. All patients underwent MRI using a 3-Tesla system (Trio Tim; Siemens, Erlangen Germany). To calculate ADC, the primary lesion was designated on the ADC map at the level of the largest tumor diameter to cover most of the lesion, avoiding cystic or necrotic components. Median and mean (±SD) pretreatment ADC were 0.713 and 0.716 ± 0.079 × 10−3 mm2/s, respectively. Univariate and multivariate analysis confirmed high pretreatment ADC was a good prognostic factor for poor local relapse-free survival and disease-free survival. Furthermore, the area under the ROC curve for prediction of local failure significantly increased when pretreatment ADC was combined with T classification (P = 0.004). Thus, pretreatment ADC might provide useful information for predicting outcome and selecting high-risk patients appropriate for more aggressive therapy. Further studies are warranted to investigate the biological basis of this observation. PMID:26184509

  11. Cloning and nucleotide sequence of wild type and a mutant histidine decarboxylase from Lactobacillus 30a.

    PubMed

    Vanderslice, P; Copeland, W C; Robertus, J D

    1986-11-15

    Prohistidine decarboxylase from Lactobacillus 30a is a protein that autoactivates to histidine decarboxylase by cleaving its peptide chain between serines 81 and 82 and converting Ser-82 to a pyruvoyl moiety. The pyruvoyl group serves as the prosthetic group for the decarboxylation reaction. We have cloned and determined the nucleotide sequence of the gene for this enzyme from a wild type strain and from a mutant with altered autoactivation properties. The nucleotide sequence modifies the previously determined amino acid sequence of the protein. A tripeptide missed in the chemical sequence is inserted, and three other amino acids show conservative changes. The activation mutant shows a single change of Gly-58 to an Asp. Sequence analysis up- and downstream from the gene suggests that histidine decarboxylase is part of a polycistronic message, and that the transcriptional promotor region is strongly homologous to those of other Gram-positive organisms.

  12. The effect of a high fat diet on pyruvate decarboxylase deficiency without central nervous system involvement.

    PubMed

    Kodama, S; Yagi, R; Ninomiya, M; Goji, K; Takahashi, T; Morishita, Y; Matsuo, T

    1983-01-01

    A nine-year-old Japanese boy with low pyruvate decarboxylase activity in fibroblasts showed no central nervous symptoms except for muscle fatigue. The pyruvate decarboxylase activities in fibroblasts of the patient and two control subjects were 0.407 +/- 0.083, 1.029 +/- 0.137 and 1.607 +/- 0.096 mumoles/g protein/30 min, respectively. The Michaelis-Menten constant (Km) was the same in the patient and controls. There was no inhibitor of pyruvate decarboxylase in the patient's fibroblasts. A high fat diet has been given to the patient for five years. At present he does not complain of any kind of muscle fatigue, except after severe exercise. Mental and physiological development of the patient are within the normal ranges. However, trials of orally administered thiamine hydrochloride or thiamine hydrochloride combined with lipoamide did not improve his muscle fatigue.

  13. 21 CFR 173.115 - Alpha-acetolactate decarboxylase (α-ALDC) enzyme preparation derived from a recombinant Bacillus...

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 3 2011-04-01 2011-04-01 false Alpha-acetolactate decarboxylase (α-ALDC) enzyme...) SECONDARY DIRECT FOOD ADDITIVES PERMITTED IN FOOD FOR HUMAN CONSUMPTION Enzyme Preparations and Microorganisms § 173.115 Alpha-acetolactate decarboxylase (α-ALDC) enzyme preparation derived from a...

  14. 21 CFR 173.115 - Alpha-acetolactate decarboxylase (α-ALDC) enzyme preparation derived from a recombinant Bacillus...

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 3 2014-04-01 2014-04-01 false Alpha-acetolactate decarboxylase (α-ALDC) enzyme... FOOD FOR HUMAN CONSUMPTION Enzyme Preparations and Microorganisms § 173.115 Alpha-acetolactate decarboxylase (α-ALDC) enzyme preparation derived from a recombinant Bacillus subtilis. The food additive...

  15. 21 CFR 173.115 - Alpha-acetolactate decarboxylase (α-ALDC) enzyme preparation derived from a recombinant Bacillus...

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 3 2012-04-01 2012-04-01 false Alpha-acetolactate decarboxylase (α-ALDC) enzyme...) SECONDARY DIRECT FOOD ADDITIVES PERMITTED IN FOOD FOR HUMAN CONSUMPTION Enzyme Preparations and Microorganisms § 173.115 Alpha-acetolactate decarboxylase (α-ALDC) enzyme preparation derived from a...

  16. 21 CFR 173.115 - Alpha-acetolactate decarboxylase (α-ALDC) enzyme preparation derived from a recombinant Bacillus...

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 3 2013-04-01 2013-04-01 false Alpha-acetolactate decarboxylase (α-ALDC) enzyme...) SECONDARY DIRECT FOOD ADDITIVES PERMITTED IN FOOD FOR HUMAN CONSUMPTION Enzyme Preparations and Microorganisms § 173.115 Alpha-acetolactate decarboxylase (α-ALDC) enzyme preparation derived from a...

  17. Ornithine Decarboxylase Activity Is Required for Prostatic Budding in the Developing Mouse Prostate

    PubMed Central

    Gamat, Melissa; Malinowski, Rita L.; Parkhurst, Linnea J.; Steinke, Laura M.; Marker, Paul C.

    2015-01-01

    The prostate is a male accessory sex gland that produces secretions in seminal fluid to facilitate fertilization. Prostate secretory function is dependent on androgens, although the mechanism by which androgens exert their effects is still unclear. Polyamines are small cationic molecules that play pivotal roles in DNA transcription, translation and gene regulation. The rate-limiting enzyme in polyamine biosynthesis is ornithine decarboxylase, which is encoded by the gene Odc1. Ornithine decarboxylase mRNA decreases in the prostate upon castration and increases upon administration of androgens. Furthermore, testosterone administered to castrated male mice restores prostate secretory activity, whereas administering testosterone and the ornithine decarboxylase inhibitor D,L-α-difluromethylornithine (DFMO) to castrated males does not restore prostate secretory activity, suggesting that polyamines are required for androgens to exert their effects. To date, no one has examined polyamines in prostate development, which is also androgen dependent. In this study, we showed that ornithine decarboxylase protein was expressed in the epithelium of the ventral, dorsolateral and anterior lobes of the adult mouse prostate. Ornithine decarboxylase protein was also expressed in the urogenital sinus (UGS) epithelium of the male and female embryo prior to prostate development, and expression continued in prostatic epithelial buds as they emerged from the UGS. Inhibiting ornithine decarboxylase using DFMO in UGS organ culture blocked the induction of prostatic buds by androgens, and significantly decreased expression of key prostate transcription factor, Nkx3.1, by androgens. DFMO also significantly decreased the expression of developmental regulatory gene Notch1. Other genes implicated in prostatic development including Sox9, Wif1 and Srd5a2 were unaffected by DFMO. Together these results indicate that Odc1 and polyamines are required for androgens to exert their effect in mediating

  18. Molecular Evolution and Functional Characterization of a Bifunctional Decarboxylase Involved in Lycopodium Alkaloid Biosynthesis1[OPEN

    PubMed Central

    Bunsupa, Somnuk; Hanada, Kousuke; Maruyama, Akira; Aoyagi, Kaori; Komatsu, Kana; Ueno, Hideki; Yamashita, Madoka; Sasaki, Ryosuke; Oikawa, Akira; Yamazaki, Mami

    2016-01-01

    Lycopodium alkaloids (LAs) are derived from lysine (Lys) and are found mainly in Huperziaceae and Lycopodiaceae. LAs are potentially useful against Alzheimer’s disease, schizophrenia, and myasthenia gravis. Here, we cloned the bifunctional lysine/ornithine decarboxylase (L/ODC), the first gene involved in LA biosynthesis, from the LA-producing plants Lycopodium clavatum and Huperzia serrata. We describe the in vitro and in vivo functional characterization of the L. clavatum L/ODC (LcL/ODC). The recombinant LcL/ODC preferentially catalyzed the decarboxylation of l-Lys over l-ornithine (l-Orn) by about 5 times. Transient expression of LcL/ODC fused with the amino or carboxyl terminus of green fluorescent protein, in onion (Allium cepa) epidermal cells and Nicotiana benthamiana leaves, showed LcL/ODC localization in the cytosol. Transgenic tobacco (Nicotiana tabacum) hairy roots and Arabidopsis (Arabidopsis thaliana) plants expressing LcL/ODC enhanced the production of a Lys-derived alkaloid, anabasine, and cadaverine, respectively, thus, confirming the function of LcL/ODC in plants. In addition, we present an example of the convergent evolution of plant Lys decarboxylase that resulted in the production of Lys-derived alkaloids in Leguminosae (legumes) and Lycopodiaceae (clubmosses). This convergent evolution event probably occurred via the promiscuous functions of the ancestral Orn decarboxylase, which is an enzyme involved in the primary metabolism of polyamine. The positive selection sites were detected by statistical analyses using phylogenetic trees and were confirmed by site-directed mutagenesis, suggesting the importance of those sites in granting the promiscuous function to Lys decarboxylase while retaining the ancestral Orn decarboxylase function. This study contributes to a better understanding of LA biosynthesis and the molecular evolution of plant Lys decarboxylase. PMID:27303024

  19. Childhood attachment and schizophrenia: the "attachment-developmental-cognitive" (ADC) hypothesis.

    PubMed

    Rajkumar, Ravi Philip

    2014-09-01

    Schizophrenia is a complex psychiatric syndrome whose exact causes remain unclear. However, current scientific consensus has highlighted the importance of neurodevelopmental and neurocognitive processes in the development of schizophrenic symptoms. Research over the past three decades, motivated by the findings of the World Health Organization's large-scale studies, has highlighted the importance of psychosocial adversities - including childhood abuse and neglect - in this disorder. In this paper, I propose a hypothesis based on John Bowlby's framework of attachment theory, which I have termed the attachment-developmental-cognitive (ADC) hypothesis. The ADC hypothesis integrates recent developments related to (1) existing models of schizophrenia, (2) studies examining the effect of attachment on brain biology and cognitive development, and (3) various known facts about the course and outcome of this disorder. In doing so, it explains how disturbed childhood attachment leads to core psychological and neurochemical abnormalities which are implicated in the genesis of schizophrenia and also affect its outcome. The ADC hypothesis compasses and expands on earlier formulations, such as the "social defeat" and "traumagenic" models, and has important implications regarding the prevention and treatment of schizophrenia. Ways of testing and refining this hypothesis are outlined as avenues for future research. Though provisional, the ADC hypothesis is entirely consistent with both biological and psychosocial research into the origins of schizophrenia.

  20. Attitude Determination and Control Subsystem (ADCS) Preparations for the EPOXI Flyby of Comet Hartley 2

    NASA Technical Reports Server (NTRS)

    Luna, Michael E.; Collins, Steven M.

    2011-01-01

    On November 4, 2010 the former "Deep Impact" spacecraft, renamed "EPOXI" for its extended mission, flew within 700km of comet 103P/Hartley 2. In July 2005, the spacecraft had previously imaged a probe impact of comet Tempel 1. The EPOXI flyby was the fifth close encounter of a spacecraft with a comet nucleus and marked the first time in history that two comet nuclei were imaged at close range with the same suite of onboard science instruments. This challenging objective made the function of the attitude determination and control subsystem (ADCS) critical to the successful execution of the EPOXI flyby.As part of the spacecraft flyby preparations, the ADCS operations team had to perform meticulous sequence reviews, implement complex spacecraft engineering and science activities and perform numerous onboard calibrations. ADCS contributions included design and execution of 10 trajectory correction maneuvers, the science calibration of the two telescopic instruments, an in-flight demonstration of high-rate turns between Earth and comet point, and an ongoing assessment of reaction wheel health. The ADCS team was also responsible for command sequences that included updates to the onboard ephemeris and sun sensor coefficients and implementation of reaction wheel assembly (RWA) de-saturations.

  1. Precision ADCS of a spinning spacecraft for the Mars Aeronomy Explorer Mission

    NASA Technical Reports Server (NTRS)

    Mungas, Greg S.; Shotwell, Robert; Gray, Andrew

    2005-01-01

    This paper discusses a precision attitude and control technique for meeting these requirements utilizing a similar architecture that was adopted for the Laboratory of Atmospheric and Space Physics (LASP) SNOE (Student Nitrous Oxide Explorer) spinning spacecraft; SNOE has been operating with its ADCS architecture in low earth orbit (LEO) for over two years.

  2. System-Level Mitigation of Undersampling ADC Nonlinearity for High-IF Radio Receivers

    NASA Astrophysics Data System (ADS)

    Vallant, Georg; Epp, Michael; Allén, Markus; Valkama, Mikko; Jondral, Friedrich K.

    2012-09-01

    Over the last years ongoing advances in ADC technology have enabled RF signals to be sampled at IF frequencies. Undersampling is nowadays employed in software-defined radio or radar receivers and offers the possibility to relieve requirements in the analog receiver partition. Unfortunately, when moving to higher IF concepts, this becomes demanding for the ADC itself, because of inherent spurious-free dynamic range (SFDR) roll-off that increases with input frequency. This fact often limits the receiver's IF placement to Nyquist zone (NZ) 2. In this work the emerging concept of Digital Assistance is pursued to give the receiver access to higher NZs while making no compromise on the SFDR. We will present and discuss post-correction results for two 16-bit high-speed converters from two different vendors at 120 and 125 MSPS, respectively. The proposed system-level post-correction decomposes nonlinearity into a static and a dynamic part. For both ADCs under investigation the degraded SFDR in higher NZs could be improved by up to 15 dB using purely digital linearization technologies, thus increasing the detectability of small signals in the presence of very strong signals or interferers. Near-identical results for both ADCs confirm the general validity of the system-level correction approach.

  3. Communication Impairment in the AIDS Dementia Complex (ADC): A Case Report

    ERIC Educational Resources Information Center

    McCabe, Patricia J.; Sheard, Christine; Code, Chris

    2008-01-01

    This article details three examinations of communication impairment over 13 months in a man with AIDS dementia complex (ADC) and compares his performance on standardised language testing with that of two control participants. He had mild language impairments as measured on standardised tests but was severely impaired in pragmatic language skills.…

  4. A low-power column-parallel ADC for high-speed CMOS image sensor

    NASA Astrophysics Data System (ADS)

    Han, Ye; Li, Quanliang; Shi, Cong; Liu, Liyuan; Wu, Nanjian

    2013-08-01

    This paper presents a 10-bit low-power column-parallel cyclic analog-to-digital converter (ADC) used for high-speed CMOS image sensor (CIS). An opamp sharing technique is used to save power and area. Correlated double sampling (CDS) circuit and programmable gain amplifier (PGA) are integrated in the ADC, which avoids stand-alone circuit blocks. An offset cancellation technique is also introduced, which reduces the column fixed-pattern noise (FPN) effectively. One single channel ADC with an area less than 0.03mm2 was implemented in a 0.18μm 1P4M CMOS image sensor process. The resolution of the proposed ADC is 10-bit, and the conversion rate is 2MS/s. The measured differential nonlinearity (DNL) and integral nonlinearity (INL) are 0.62 LSB and 2.1 LSB together with CDS, respectively. The power consumption from 1.8V supply is only 0.36mW.

  5. Cloning and sequencing of pyruvate decarboxylase (PDC) genes from bacteria and uses therefor

    DOEpatents

    Maupin-Furlow, Julie A [Gainesville, FL; Talarico, Lee Ann [Gainesville, FL; Raj, Krishnan Chandra [Tamil Nadu, IN; Ingram, Lonnie O [Gainesville, FL

    2008-02-05

    The invention provides isolated nucleic acids molecules which encode pyruvate decarboxylase enzymes having improved decarboxylase activity, substrate affinity, thermostability, and activity at different pH. The nucleic acids of the invention also have a codon usage which allows for high expression in a variety of host cells. Accordingly, the invention provides recombinant expression vectors containing such nucleic acid molecules, recombinant host cells comprising the expression vectors, host cells further comprising other ethanologenic enzymes, and methods for producing useful substances, e.g., acetaldehyde and ethanol, using such host cells.

  6. ADC texture—An imaging biomarker for high-grade glioma?

    SciTech Connect

    Brynolfsson, Patrik; Hauksson, Jón; Karlsson, Mikael; Garpebring, Anders; Nyholm, Tufve; Nilsson, David; Trygg, Johan; Henriksson, Roger; Birgander, Richard; Asklund, Thomas

    2014-10-15

    Purpose: Survival for high-grade gliomas is poor, at least partly explained by intratumoral heterogeneity contributing to treatment resistance. Radiological evaluation of treatment response is in most cases limited to assessment of tumor size months after the initiation of therapy. Diffusion-weighted magnetic resonance imaging (MRI) and its estimate of the apparent diffusion coefficient (ADC) has been widely investigated, as it reflects tumor cellularity and proliferation. The aim of this study was to investigate texture analysis of ADC images in conjunction with multivariate image analysis as a means for identification of pretreatment imaging biomarkers. Methods: Twenty-three consecutive high-grade glioma patients were treated with radiotherapy (2 Gy/60 Gy) with concomitant and adjuvant temozolomide. ADC maps and T1-weighted anatomical images with and without contrast enhancement were collected prior to treatment, and (residual) tumor contrast enhancement was delineated. A gray-level co-occurrence matrix analysis was performed on the ADC maps in a cuboid encapsulating the tumor in coronal, sagittal, and transversal planes, giving a total of 60 textural descriptors for each tumor. In addition, similar examinations and analyses were performed at day 1, week 2, and week 6 into treatment. Principal component analysis (PCA) was applied to reduce dimensionality of the data, and the five largest components (scores) were used in subsequent analyses. MRI assessment three months after completion of radiochemotherapy was used for classifying tumor progression or regression. Results: The score scatter plots revealed that the first, third, and fifth components of the pretreatment examinations exhibited a pattern that strongly correlated to survival. Two groups could be identified: one with a median survival after diagnosis of 1099 days and one with 345 days, p = 0.0001. Conclusions: By combining PCA and texture analysis, ADC texture characteristics were identified, which seems

  7. The metalloregulatory zinc site in Streptococcus pneumoniae AdcR, a zinc-activated MarR-family repressor

    PubMed Central

    Reyes-Caballero, Hermes; Guerra, Alfredo J.; Jacobsen, Faith E.; Kazmierczak, Krystyna M.; Cowart, Darin; Koppolu, Uma Mahendra Kumar; Scott, Robert A.; Winkler, Malcolm E.; Giedroc, David P.

    2010-01-01

    Streptococcus pneumoniae D39 AdcR (adhesin competence repressor) is the first metal-sensing member of the MarR (multiple antibiotic resistance repressor) family to be characterized. Expression profiling with a ΔadcR strain grown in liquid culture (brain heart infusion; BHI) under microaerobic conditions reveals upregulation of 13 genes including adcR and adcCBA, encoding a high affinity ABC uptake system for zinc, and genes encoding cell-surface zinc-binding pneumococcal histidine triad (Pht) proteins and AdcAII (Lmb, laminin binding). The ΔadcR, H108Q and H112Q adcR mutant allelic strains grown in 0.2 mM Zn(II) exhibit a slow-growth phenotype and a ≈2-fold increase in cell-associated Zn(II). Apo- and Zn(II)-bound AdcR are homodimers in solution and binding to a 28-mer DNA containing an adc operator is strongly stimulated by Zn(II) with KDNA-Zn = 2.4 ×108 M−1 (pH 6.0, 0.2 M NaCl, 25 °C). AdcR binds two Zn(II) per dimer, with step-wise Zn(II) affinities KZn1 and KZn2 of ≥109 M−1 at pH 6.0 and ≥1012 M−1 at pH 8.0. X-ray absorption spectroscopy (XAS) of the high affinity site reveals a pentacoordinate N/O complex and no cysteine coordination, the latter finding corroborated by wild-type-like functional properties of C30A AdcR. Alanine substitution of conserved residues His42 in the DNA binding domain, and His108 and His112 in the C-terminal regulatory domain, abolish high affinity Zn(II) binding and greatly reduce Zn(II)-activated binding to DNA. NMR studies reveal that these mutants adopt the same folded conformation as dimeric wild-type apo AdcR, but fail to conformationally switch upon Zn(II) binding. These studies clearly identify His42, His108 and H112 as metalloregulatory zinc ligands in S. pneumoniae AdcR. PMID:20804771

  8. Phosphorylation of Ser-204 and Tyr-405 in human malonyl-CoA decarboxylase expressed in silkworm Bombyx mori regulates catalytic decarboxylase activity.

    PubMed

    Hwang, In-Wook; Makishima, Yu; Suzuki, Tomohiro; Kato, Tatsuya; Park, Sungjo; Terzic, Andre; Chung, Shin-Kyo; Park, Enoch Y

    2015-11-01

    Decarboxylation of malonyl-CoA to acetyl-CoA by malonyl-CoA decarboxylase (MCD; EC 4.1.1.9) is a vital catalytic reaction of lipid metabolism. While it is established that phosphorylation of MCD modulates the enzymatic activity, the specific phosphorylation sites associated with the catalytic function have not been documented due to lack of sufficient production of MCD with proper post-translational modifications. Here, we used the silkworm-based Bombyx mori nucleopolyhedrovirus (BmNPV) bacmid system to express human MCD (hMCD) and mapped phosphorylation effects on enzymatic function. Purified MCD from silkworm displayed post-translational phosphorylation and demonstrated coherent enzymatic activity with high yield (-200 μg/silkworm). Point mutations in putative phosphorylation sites, Ser-204 or Tyr-405 of hMCD, identified by bioinformatics and proteomics analyses reduced the catalytic activity, underscoring the functional significance of phosphorylation in modulating decarboxylase-based catalysis. Identified phosphorylated residues are distinct from the decarboxylation catalytic site, implicating a phosphorylation-induced global conformational change of MCD as responsible in altering catalytic function. We conclude that phosphorylation of Ser-204 and Tyr-405 regulates the decarboxylase function of hMCD leveraging the silkworm-based BmNPV bacmid expression system that offers a fail-safe eukaryotic production platform implementing proper post-translational modification such as phosphorylation.

  9. Treatment with the C5a receptor antagonist ADC-1004 reduces myocardial infarction in a porcine ischemia-reperfusion model

    PubMed Central

    2010-01-01

    Background Polymorphonuclear neutrophils, stimulated by the activated complement factor C5a, have been implicated in cardiac ischemia/reperfusion injury. ADC-1004 is a competitive C5a receptor antagonist that has been shown to inhibit complement related neutrophil activation. ADC-1004 shields the neutrophils from C5a activation before they enter the reperfused area, which could be a mechanistic advantage compared to previous C5a directed reperfusion therapies. We investigated if treatment with ADC-1004, according to a clinically applicable protocol, would reduce infarct size and microvascular obstruction in a large animal myocardial infarct model. Methods In anesthetized pigs (42-53 kg), a percutaneous coronary intervention balloon was inflated in the left anterior descending artery for 40 minutes, followed by 4 hours of reperfusion. Twenty minutes after balloon inflation the pigs were randomized to an intravenous bolus administration of ADC-1004 (175 mg, n = 8) or saline (9 mg/ml, n = 8). Area at risk (AAR) was evaluated by ex vivo SPECT. Infarct size and microvascular obstruction were evaluated by ex vivo MRI. The observers were blinded to the treatment at randomization and analysis. Results ADC-1004 treatment reduced infarct size by 21% (ADC-1004: 58.3 ± 3.4 vs control: 74.1 ± 2.9%AAR, p = 0.007). Microvascular obstruction was similar between the groups (ADC-1004: 2.2 ± 1.2 vs control: 5.3 ± 2.5%AAR, p = 0.23). The mean plasma concentration of ADC-1004 was 83 ± 8 nM at sacrifice. There were no significant differences between the groups with respect to heart rate, mean arterial pressure, cardiac output and blood-gas data. Conclusions ADC-1004 treatment reduces myocardial ischemia-reperfusion injury and represents a novel treatment strategy of myocardial infarct with potential clinical applicability. PMID:20875134

  10. Value of ADC measurements for nodal staging after chemoradiation in locally advanced rectal cancer—a per lesion validation study

    PubMed Central

    Lambregts, Doenja M. J.; Maas, Monique; Riedl, Robert G.; Bakers, Frans C. H.; Verwoerd, Jan L.; Kessels, Alfons G. H.; Lammering, Guido; Boetes, Carla; Beets, Geerard L.

    2010-01-01

    Objectives To evaluate the performance of diffusion-weighted MRI (DWI) in addition to T2-weighted (T2W) MRI for nodal restaging after chemoradiation in rectal cancer. Methods Thirty patients underwent chemoradiation followed by MRI (1.5 T) and surgery. Imaging consisted of T2W-MRI and DWI (b0, 500, 1000). On T2W-MRI, nodes were scored as benign/malignant by two independent readers (R1, R2). Mean apparent diffusion coefficient (ADC) was measured for each node. Diagnostic performance was compared for T2W-MRI, ADC and T2W+ADC, using a per lesion histological validation. Results ADC was higher for the malignant nodes (1.43 ± 0.38 vs 1.19 ± 0.27 *10−3 mm2/s, p < 0.001). Area under the ROC curve/sensitivity/specificity were 0.88/65%/93% (R1) and 0.95/71%/91% (R2) using T2W-MRI; 0.66/53%/82% using ADC (mean of two readers); and 0.91/56%/98% (R1) and 0.96/56%/99% (R2) using T2W+ADC. There was no significant difference between T2W-MRI and T2W+ADC. Interobserver reproducibility was good for T2W-MRI (κ0.73) and ADC (intraclass correlation coefficient 0.77). Conclusions After chemoradiation, ADC measurements may have potential for nodal characterisation, but DWI on its own is not reliable. Addition of DWI to T2W-MRI does not improve accuracy and T2W-MRI is already sufficiently accurate. PMID:20730540

  11. Structural and Mechanistic Studies on Klebsiella pneumoniae 2-Oxo-4-hydroxy-4-carboxy-5-ureidoimidazoline Decarboxylase

    SciTech Connect

    French, Jarrod B.; Ealick, Steven E.

    2010-11-12

    The stereospecific oxidative degradation of uric acid to (S)-allantoin was recently shown to proceed via three enzymatic steps. The final conversion is a decarboxylation of the unstable intermediate 2-oxo-4-hydroxy-4-carboxy-5-ureidoimidazoline (OHCU) and is catalyzed by OHCU decarboxylase. Here we present the structures of Klebsiella pneumoniae OHCU decarboxylase in unliganded form and with bound allantoin. These structures provide evidence that ligand binding organizes the active site residues for catalysis. Modeling of the substrate and intermediates provides additional support for this hypothesis. In addition we characterize the steady state kinetics of this enzyme and report the first OHCU decarboxylase inhibitor, allopurinol, a structural isomer of hypoxanthine. This molecule is a competitive inhibitor of K. pneumoniae OHCU decarboxylase with a K{sub i} of 30 {+-} 2 {micro}m. Circular dichroism measurements confirm structural observations that this inhibitor disrupts the necessary organization of the active site. Our structural and biochemical studies also provide further insights into the mechanism of catalysis of OHCU decarboxylation.

  12. Detection and transfer of the glutamate decarboxylase gene in Streptococcus thermophilus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    GABA (gamma-aminobutyric acid) is generated from glutamate by the action of glutamic acid decarboxylase (GAD) and characterized by hypotensive, diuretic and tranquilizing effects in humans and animals. The production of GABA by lactic acid starter bacteria would enhance the functionality of fermen...

  13. Molecular analysis of the glutamate decarboxylase locus in Streptococcus thermophilus ST110

    Technology Transfer Automated Retrieval System (TEKTRAN)

    GABA ('-aminobutyric acid) is generated from glutamate by the action of glutamic acid decarboxylase (GAD) and characterized by hypotensive, diuretic and tranquilizing effects in humans and animals. The production of GABA by lactic acid starter bacteria would enhance the functionality of fermented da...

  14. Inhibition of pyruvate decarboxylase from Z. mobilis by novel analogues of thiamine pyrophosphate: investigating pyrophosphate mimics.

    PubMed

    Erixon, Karl M; Dabalos, Chester L; Leeper, Finian J

    2007-03-07

    Replacement of the thiazolium ring of thiamine pyrophosphate with a triazole gives extremely potent inhibitors of pyruvate decarboxylase from Z. mobilis, with K(I) values down to 20 pM; this system was used to explore pyrophosphate mimics and several effective analogues were discovered.

  15. The Degradation of 14C-Glutamic Acid by L-Glutamic Acid Decarboxylase.

    ERIC Educational Resources Information Center

    Dougherty, Charles M; Dayan, Jean

    1982-01-01

    Describes procedures and semi-micro reaction apparatus (carbon dioxide trap) to demonstrate how a particular enzyme (L-Glutamic acid decarboxylase) may be used to determine the site or sites of labeling in its substrate (carbon-14 labeled glutamic acid). Includes calculations, solutions, and reagents used. (Author/SK)

  16. Presentation of opsoclonus myoclonus ataxia syndrome with glutamic acid decarboxylase antibodies.

    PubMed

    Bhandari, Hanul Srinivas

    2012-08-08

    In this rare case, the patient presented with opsoclonus, myoclonus and ataxia. Serological and imaging studies revealed high glutamic acid decarboxylase antibody (GAD-Ab) levels. High-dose corticosteroids were of no benefit and subsequent intravenous immunoglobulin (IVIg) administration proved resolution of the condition. Levetiracetam proved useful in symptomatically controlling the myoclonus. Follow-up GAD-Ab levels were within normal limits.

  17. Analysis and comparison of CVS-ADC approaches up to third order for the calculation of core-excited states

    NASA Astrophysics Data System (ADS)

    Wenzel, Jan; Holzer, Andre; Wormit, Michael; Dreuw, Andreas

    2015-06-01

    The extended second order algebraic-diagrammatic construction (ADC(2)-x) scheme for the polarization operator in combination with core-valence separation (CVS) approximation is well known to be a powerful quantum chemical method for the calculation of core-excited states and the description of X-ray absorption spectra. For the first time, the implementation and results of the third order approach CVS-ADC(3) are reported. Therefore, the CVS approximation has been applied to the ADC(3) working equations and the resulting terms have been implemented efficiently in the adcman program. By treating the α and β spins separately from each other, the unrestricted variant CVS-UADC(3) for the treatment of open-shell systems has been implemented as well. The performance and accuracy of the CVS-ADC(3) method are demonstrated with respect to a set of small and middle-sized organic molecules. Therefore, the results obtained at the CVS-ADC(3) level are compared with CVS-ADC(2)-x values as well as experimental data by calculating complete basis set limits. The influence of basis sets is further investigated by employing a large set of different basis sets. Besides the accuracy of core-excitation energies and oscillator strengths, the importance of cartesian basis functions and the treatment of orbital relaxation effects are analyzed in this work as well as computational timings. It turns out that at the CVS-ADC(3) level, the results are not further improved compared to CVS-ADC(2)-x and experimental data, because the fortuitous error compensation inherent in the CVS-ADC(2)-x approach is broken. While CVS-ADC(3) overestimates the core excitation energies on average by 0.61% ± 0.31%, CVS-ADC(2)-x provides an averaged underestimation of -0.22% ± 0.12%. Eventually, the best agreement with experiments can be achieved using the CVS-ADC(2)-x method in combination with a diffuse cartesian basis set at least at the triple-ζ level.

  18. Analysis and comparison of CVS-ADC approaches up to third order for the calculation of core-excited states.

    PubMed

    Wenzel, Jan; Holzer, Andre; Wormit, Michael; Dreuw, Andreas

    2015-06-07

    The extended second order algebraic-diagrammatic construction (ADC(2)-x) scheme for the polarization operator in combination with core-valence separation (CVS) approximation is well known to be a powerful quantum chemical method for the calculation of core-excited states and the description of X-ray absorption spectra. For the first time, the implementation and results of the third order approach CVS-ADC(3) are reported. Therefore, the CVS approximation has been applied to the ADC(3) working equations and the resulting terms have been implemented efficiently in the adcman program. By treating the α and β spins separately from each other, the unrestricted variant CVS-UADC(3) for the treatment of open-shell systems has been implemented as well. The performance and accuracy of the CVS-ADC(3) method are demonstrated with respect to a set of small and middle-sized organic molecules. Therefore, the results obtained at the CVS-ADC(3) level are compared with CVS-ADC(2)-x values as well as experimental data by calculating complete basis set limits. The influence of basis sets is further investigated by employing a large set of different basis sets. Besides the accuracy of core-excitation energies and oscillator strengths, the importance of cartesian basis functions and the treatment of orbital relaxation effects are analyzed in this work as well as computational timings. It turns out that at the CVS-ADC(3) level, the results are not further improved compared to CVS-ADC(2)-x and experimental data, because the fortuitous error compensation inherent in the CVS-ADC(2)-x approach is broken. While CVS-ADC(3) overestimates the core excitation energies on average by 0.61% ± 0.31%, CVS-ADC(2)-x provides an averaged underestimation of -0.22% ± 0.12%. Eventually, the best agreement with experiments can be achieved using the CVS-ADC(2)-x method in combination with a diffuse cartesian basis set at least at the triple-ζ level.

  19. A 1.8 V low-power 14-bit 20 Msps ADC with 11.2 ENOB

    NASA Astrophysics Data System (ADS)

    Hua, Cai

    2012-11-01

    This paper describes the design of a 14-bit 20 Msps analog-to-digital converter (ADC), implemented in 0.18 μm CMOS technology, achieving 11.2 effective number of bits at Nyquist rate. An improved SHA-less structure and op-amp sharing technique is adopted to significantly reduce the power. The proposed ADC consumes only 166 mW under 1.8 V supply. A fast background calibration is utilized to ensure the overall ADC linearity.

  20. In-pixel conversion with a 10 bit SAR ADC for next generation X-ray FELs

    NASA Astrophysics Data System (ADS)

    Lodola, L.; Batignani, G.; Benkechkache, M. A.; Bettarini, S.; Casarosa, G.; Comotti, D.; Dalla Betta, G. F.; Fabris, L.; Forti, F.; Grassi, M.; Latreche, S.; Malcovati, P.; Manghisoni, M.; Mendicino, R.; Morsani, F.; Paladino, A.; Pancheri, L.; Paoloni, E.; Ratti, L.; Re, V.; Rizzo, G.; Traversi, G.; Vacchi, C.; Verzellesi, G.; Xu, H.

    2016-07-01

    This work presents the design of an interleaved Successive Approximation Register (SAR) ADC, part of the readout channel for the PixFEL detector. The PixFEL project aims at substantially advancing the state-of-the-art in the field of 2D X-ray imaging for applications at the next generation Free Electron Laser (FEL) facilities. For this purpose, the collaboration is developing the fundamental microelectronic building blocks for the readout channel. This work focuses on the design of the ADC carried out in a 65 nm CMOS technology. To obtain a good tradeoff between power consumption, conversion speed and area occupation, an interleaved SAR ADC architecture was adopted.

  1. The enzymatic activities of the Escherichia coli basic aliphatic amino acid decarboxylases exhibit a pH zone of inhibition.

    PubMed

    Kanjee, Usheer; Gutsche, Irina; Ramachandran, Shaliny; Houry, Walid A

    2011-11-01

    The stringent response regulator ppGpp has recently been shown by our group to inhibit the Escherichia coli inducible lysine decarboxylase, LdcI. As a follow-up to this observation, we examined the mechanisms that regulate the activities of the other four E. coli enzymes paralogous to LdcI: the constitutive lysine decarboxylase LdcC, the inducible arginine decarboxylase AdiA, the inducible ornithine decarboxylase SpeF, and the constitutive ornithine decarboxylase SpeC. LdcC and SpeC are involved in cellular polyamine biosynthesis, while LdcI, AdiA, and SpeF are involved in the acid stress response. Multiple mechanisms of regulation were found for these enzymes. In addition to LdcI, LdcC and SpeC were found to be inhibited by ppGpp; AdiA activity was found to be regulated by changes in oligomerization, while SpeF and SpeC activities were regulated by GTP. These findings indicate the presence of multiple mechanisms regulating the activity of this important family of decarboxylases. When the enzyme inhibition profiles are analyzed in parallel, a "zone of inhibition" between pH 6 and pH 8 is observed. Hence, the data suggest that E. coli utilizes multiple mechanisms to ensure that these decarboxylases remain inactive around neutral pH possibly to reduce the consumption of amino acids at this pH.

  2. Monomeric S-adenosylmethionine decarboxylase from plants provides an alternative to putrescine stimulation.

    PubMed

    Bennett, Eric M; Ekstrom, Jennifer L; Pegg, Anthony E; Ealick, Steven E

    2002-12-10

    S-Adenosylmethionine decarboxylase has been implicated in cell growth and differentiation and is synthesized as a proenzyme, which undergoes autocatalytic cleavage to generate an active site pyruvoyl group. In mammals, S-adenosylmethionine decarboxylase is active as a dimer in which each protomer contains one alpha subunit and one beta subunit. In many higher organisms, autocatalysis and decarboxylation are stimulated by putrescine, which binds in a buried site containing numerous negatively charged residues. In contrast, plant S-adenosylmethionine decarboxylases are fully active in the absence of putrescine, with rapid autocatalysis that is not stimulated by putrescine. We have determined the structure of the S-adenosylmethionine decarboxylase from potato, Solanum tuberosum, to 2.3 A resolution. Unlike the previously determined human enzyme structure, the potato enzyme is a monomer in the crystal structure. Ultracentrifugation studies show that the potato enzyme is also a monomer under physiological conditions, with a weak self-association constant of 6.5 x 10(4) M(-)(1) for the monomer-dimer association. Although the potato enzyme contains most of the buried charged residues that make up the putrescine binding site in the human enzyme, there is no evidence for a putrescine binding site in the potato enzyme. Instead, several amino acid substitutions, including Leu13/Arg18, Phe111/Arg114, Asp174/Val181, and Phe285/His294 (human/potato), provide side chains that mimic the role of putrescine in the human enzyme. In the potato enzyme, the positively charged residues form an extensive network of hydrogen bonds bridging a cluster of highly conserved negatively charged residues and the active site, including interactions with the catalytic residues Glu16 and His249. The results explain the constitutively high activity of plant S-adenosylmethionine decarboxylases in the absence of putrescine and are consistent with previously proposed models for how putrescine together

  3. Substrate specificity of thiamine pyrophosphate-dependent 2-oxo-acid decarboxylases in Saccharomyces cerevisiae.

    PubMed

    Romagnoli, Gabriele; Luttik, Marijke A H; Kötter, Peter; Pronk, Jack T; Daran, Jean-Marc

    2012-11-01

    Fusel alcohols are precursors and contributors to flavor and aroma compounds in fermented beverages, and some are under investigation as biofuels. The decarboxylation of 2-oxo acids is a key step in the Ehrlich pathway for fusel alcohol production. In Saccharomyces cerevisiae, five genes share sequence similarity with genes encoding thiamine pyrophosphate-dependent 2-oxo-acid decarboxylases (2ODCs). PDC1, PDC5, and PDC6 encode differentially regulated pyruvate decarboxylase isoenzymes; ARO10 encodes a 2-oxo-acid decarboxylase with broad substrate specificity, and THI3 has not yet been shown to encode an active decarboxylase. Despite the importance of fusel alcohol production in S. cerevisiae, the substrate specificities of these five 2ODCs have not been systematically compared. When the five 2ODCs were individually overexpressed in a pdc1Δ pdc5Δ pdc6Δ aro10Δ thi3Δ strain, only Pdc1, Pdc5, and Pdc6 catalyzed the decarboxylation of the linear-chain 2-oxo acids pyruvate, 2-oxo-butanoate, and 2-oxo-pentanoate in cell extracts. The presence of a Pdc isoenzyme was also required for the production of n-propanol and n-butanol in cultures grown on threonine and norvaline, respectively, as nitrogen sources. These results demonstrate the importance of pyruvate decarboxylases in the natural production of n-propanol and n-butanol by S. cerevisiae. No decarboxylation activity was found for Thi3 with any of the substrates tested. Only Aro10 and Pdc5 catalyzed the decarboxylation of the aromatic substrate phenylpyruvate, with Aro10 showing superior kinetic properties. Aro10, Pdc1, Pdc5, and Pdc6 exhibited activity with all branched-chain and sulfur-containing 2-oxo acids tested but with markedly different decarboxylation kinetics. The high affinity of Aro10 identified it as a key contributor to the production of branched-chain and sulfur-containing fusel alcohols.

  4. Substrate Specificity of Thiamine Pyrophosphate-Dependent 2-Oxo-Acid Decarboxylases in Saccharomyces cerevisiae

    PubMed Central

    Romagnoli, Gabriele; Luttik, Marijke A. H.; Kötter, Peter; Pronk, Jack T.

    2012-01-01

    Fusel alcohols are precursors and contributors to flavor and aroma compounds in fermented beverages, and some are under investigation as biofuels. The decarboxylation of 2-oxo acids is a key step in the Ehrlich pathway for fusel alcohol production. In Saccharomyces cerevisiae, five genes share sequence similarity with genes encoding thiamine pyrophosphate-dependent 2-oxo-acid decarboxylases (2ODCs). PDC1, PDC5, and PDC6 encode differentially regulated pyruvate decarboxylase isoenzymes; ARO10 encodes a 2-oxo-acid decarboxylase with broad substrate specificity, and THI3 has not yet been shown to encode an active decarboxylase. Despite the importance of fusel alcohol production in S. cerevisiae, the substrate specificities of these five 2ODCs have not been systematically compared. When the five 2ODCs were individually overexpressed in a pdc1Δ pdc5Δ pdc6Δ aro10Δ thi3Δ strain, only Pdc1, Pdc5, and Pdc6 catalyzed the decarboxylation of the linear-chain 2-oxo acids pyruvate, 2-oxo-butanoate, and 2-oxo-pentanoate in cell extracts. The presence of a Pdc isoenzyme was also required for the production of n-propanol and n-butanol in cultures grown on threonine and norvaline, respectively, as nitrogen sources. These results demonstrate the importance of pyruvate decarboxylases in the natural production of n-propanol and n-butanol by S. cerevisiae. No decarboxylation activity was found for Thi3 with any of the substrates tested. Only Aro10 and Pdc5 catalyzed the decarboxylation of the aromatic substrate phenylpyruvate, with Aro10 showing superior kinetic properties. Aro10, Pdc1, Pdc5, and Pdc6 exhibited activity with all branched-chain and sulfur-containing 2-oxo acids tested but with markedly different decarboxylation kinetics. The high affinity of Aro10 identified it as a key contributor to the production of branched-chain and sulfur-containing fusel alcohols. PMID:22904058

  5. Development of multichannel analyzer using sound card ADC for nuclear spectroscopy system

    NASA Astrophysics Data System (ADS)

    Ibrahim, Maslina Mohd; Yussup, Nolida; Lombigit, Lojius; Rahman, Nur Aira Abdul; Jaafar, Zainudin

    2014-02-01

    This paper describes the development of Multi-Channel Analyzer (MCA) using sound card analogue to digital converter (ADC) for nuclear spectroscopy system. The system was divided into a hardware module and a software module. Hardware module consist of detector NaI (Tl) 2" by 2", Pulse Shaping Amplifier (PSA) and a build in ADC chip from readily available in any computers' sound system. The software module is divided into two parts which are a pre-processing of raw digital input and the development of the MCA software. Band-pass filter and baseline stabilization and correction were implemented for the pre-processing. For the MCA development, the pulse height analysis method was used to process the signal before displaying it using histogram technique. The development and tested result for using the sound card as an MCA are discussed.

  6. Two-Dimensional Spatio-Temporal Signal Processing for Dispersion Compensation in Time-Stretched ADC

    NASA Astrophysics Data System (ADS)

    Tarighat, Alireza; Gupta, Shalabh; Sayed, Ali H.; Jalali, Bahram

    2007-06-01

    Time-stretched analog-to-digital converters (ADCs) have offered revolutionary enhancements in the performance of electronic converters by reducing the signal bandwidth prior to digitization. An inherent limitation of the time-stretched ADC is the frequency-selective response of the optical system that reduces the effective number of bits for ultrawideband signals. This paper proposes a solution based on spatio-temporal digital processing. The digital algorithm exploits the optical phase diversity to create a flat RF frequency response, even when the system's transfer function included deep nulls within the signal spectrum. For a 10× time-stretch factor with a 10-GHz input signal, simulations show that the proposed solution increases the overall achievable signal-to-noise-and-distortion ratio to 52 dB in the presence of linear distortions. The proposed filter can be used to mitigate the dispersion penalty in other fiber optic applications as well.

  7. A low power level-crossing ADC for wearable wireless ECG sensors.

    PubMed

    Zhenzhen Tian; Rendong Ying; Peilin Liu; Guoxing Wang; Yong Lian

    2016-08-01

    Ultra-low power consumption is desired in most wearable biomedical devices. The event-driven based Analog-to-Digital converter (ADC) could be an excellent candidate for low power system because of the reduction in sampling points for biosignals. In the existing event-driven based ADC architectures, two or more high precision comparators are utilized to sample the input signal. In this paper, we propose a new scheme utilizing only one high precision comparator to genertate samples with the assistance of a low precision one. From the Matlab simulations on real ECG signal, it is shown that around 25% reduction on the number of samples can be be achieved compared with the Nyquist sampling scheme.

  8. The Infrared Imaging Spectrograph (IRIS) for TMT: the ADC optical design

    NASA Astrophysics Data System (ADS)

    Phillips, Andrew C.; Suzuki, Ryuji; Larkin, James E.; Moore, Anna M.; Hayano, Yutaka; Tsuzuki, Toshihiro; Wright, Shelley A.

    2016-08-01

    We present the current optical design for the IRIS Atmospheric Dispersion Corrector (ADC). The ADC is designed for residual dispersions less than 1 mas across a given passband at elevations of 25 degrees. Since the last report, the area of the IRIS Imager has increased by a factor of four, and the pupil size has increased from 75 to 90mm, both of which contribute to challenges with the design. Several considerations have led to the current design: residual dispersion, amount of introduced distortion, glass transmission, glass availability, and pupil displacement. In particular, it was found that there are significant distortions that appear (two different components) that can lead to image blur over long exposures. Also, pupil displacement increases the wave front error at the imager focus. We discuss these considerations, discuss the compromises, and present the final design choice and expected performance.

  9. Experiments and FEM simulations of fracture behaviors for ADC12 aluminum alloy under impact load

    NASA Astrophysics Data System (ADS)

    Hu, Yumei; Xiao, Yue; Jin, Xiaoqing; Zheng, Haoran; Zhou, Yinge; Shao, Jinhua

    2016-11-01

    Using the combination of experiment and simulation, the fracture behavior of the brittle metal named ADC12 aluminum alloy was studied. Five typical experiments were carried out on this material, with responding data collected on different stress states and dynamic strain rates. Fractographs revealed that the morphologies of fractured specimen under several rates showed different results, indicating that the fracture was predominantly a brittle one in nature. Simulations of the fracture processes of those specimens were conducted by Finite Element Method, whilst consistency was observed between simulations and experiments. In simulation, the Johnson- Cook model was chosen to describe the damage development and to predict the failure using parameters determined from those experimental data. Subsequently, an ADC12 engine mount bracket crashing simulation was conducted and the results indicated good agreement with the experiments. The accordance showed that our research can provide an accurate description for the deforming and fracture processes of the studied alloy.

  10. Development of multichannel analyzer using sound card ADC for nuclear spectroscopy system

    SciTech Connect

    Ibrahim, Maslina Mohd; Yussup, Nolida; Lombigit, Lojius; Rahman, Nur Aira Abdul; Jaafar, Zainudin

    2014-02-12

    This paper describes the development of Multi-Channel Analyzer (MCA) using sound card analogue to digital converter (ADC) for nuclear spectroscopy system. The system was divided into a hardware module and a software module. Hardware module consist of detector NaI (Tl) 2” by 2”, Pulse Shaping Amplifier (PSA) and a build in ADC chip from readily available in any computers’ sound system. The software module is divided into two parts which are a pre-processing of raw digital input and the development of the MCA software. Band-pass filter and baseline stabilization and correction were implemented for the pre-processing. For the MCA development, the pulse height analysis method was used to process the signal before displaying it using histogram technique. The development and tested result for using the sound card as an MCA are discussed.

  11. A 13-bit Noise Shaping SAR-ADC with Dual-Polarity Digital Calibration.

    PubMed

    Park, Hangue; Ghovanloo, Maysam

    2013-06-01

    We present a new noise shaping method and a dual polarity calibration technique suited for successive approximation register type analog to digital converters (SAR-ADC). Noise is pushed to higher frequencies with the noise shaping by adding a switched capacitor. The SAR capacitor array mismatch has been compensated by the dual-polarity digital calibration with minimum circuit overhead. A proof-of-concept prototype SAR-ADC using the proposed techniques has been fabricated in a 0.5-μm standard CMOS technology. It achieves 67.7 dB SNDR at 62.5 kHz sampling frequency, while consuming 38.3μW power with 1.8 V supply.

  12. Extension of the ADC Charge-Collection Model to Include Multiple Junctions

    NASA Technical Reports Server (NTRS)

    Edmonds, Larry D.

    2011-01-01

    The ADC model is a charge-collection model derived for simple p-n junction silicon diodes having a single reverse-biased p-n junction at one end and an ideal substrate contact at the other end. The present paper extends the model to include multiple junctions, and the goal is to estimate how collected charge is shared by the different junctions.

  13. A fast, low-power, 6-bit SAR ADC for readout of strip detectors in the LHCb Upgrade experiment

    NASA Astrophysics Data System (ADS)

    Firlej, M.; Fiutowski, T.; Idzik, M.; Moron, J.; Swientek, K.

    2014-07-01

    The readout of silicon strip sensors in the upgraded Tracker System of Large Hadron Collider beauty (LHCb) experiment will require a novel complex Application Specific Integrated Circuit (ASIC). The ASIC will extract and digitise analogue signal from the sensor and subsequently will perform digital processing and serial data transmission. One of the key processing blocks, placed in each channel, will be an Analogue to Digital Converter (ADC). A prototype of fast, low-power 6-bit Successive Approximation Register (SAR) ADC was designed, fabricated and tested. The measurements of ADC prototypes confirmed simulation results showing excellent overall performance. In particular, very good resolution with Effective Number Of Bits (ENOB) 5.85 was obtained together with very low power consumption of 0.35 mW at 40 MS/s sampling rate. The results of the performed static and dynamic measurements confirm excellent ADC operation for higher sampling rates up to 80 MS/s.

  14. From Anthramycin to Pyrrolobenzodiazepine (PBD)‐Containing Antibody–Drug Conjugates (ADCs)

    PubMed Central

    Mantaj, Julia; Jackson, Paul J. M.; Rahman, Khondaker M.

    2016-01-01

    Abstract The pyrrolo[2,1‐c][1,4]benzodiazepines (PBDs) are a family of sequence‐selective DNA minor‐groove binding agents that form a covalent aminal bond between their C11‐position and the C2‐NH2 groups of guanine bases. The first example of a PBD monomer, the natural product anthramycin, was discovered in the 1960s, and the best known PBD dimer, SJG‐136 (also known as SG2000, NSC 694501 or BN2629), was synthesized in the 1990s and has recently completed Phase II clinical trials in patients with leukaemia and ovarian cancer. More recently, PBD dimer analogues are being attached to tumor‐targeting antibodies to create antibody–drug conjugates (ADCs), a number of which are now in clinical trials, with many others in pre‐clinical development. This Review maps the development from anthramycin to the first PBD dimers, and then to PBD‐containing ADCs, and explores both structure–activity relationships (SARs) and the biology of PBDs, and the strategies for their use as payloads for ADCs. PMID:27862776

  15. PEALL4: a 4-channel, 12-bit, 40-MSPS, Power Efficient and Low Latency SAR ADC

    NASA Astrophysics Data System (ADS)

    Rarbi, F.; Dzahini, D.; Gallin-Martel, L.; Bouvier, J.; Zeloufi, M.; Trocme, B.; Gabaldon Ruiz, C.

    2015-01-01

    The PEALL4 chip is a Power Efficient And Low Latency 4-channels, 12-bit and 40-MSPS successive approximation register (SAR) ADC. It was designed featuring a very short latency time in the context of ATLAS Liquid Argon Calorimeter phase I upgrade. Moreover this design could be a good option for ATLAS phase II and other High Energy Physics (HEP) projects. The full functionality of the converter is achieved by an embedded high-speed clock frequency conversion generated by the ADC itself. The design and testing results of the PEALL4 chip implemented in a commercial 130nm CMOS process are presented. The size of this 4-channel ADC with embedded voltage references and sLVS output serializer is 2.8x3.4 mm2. The chip presents a short latency time less than 25 ns defined from the very beginning of the sampling to the last conversion bit made available. A total power consumption below 27mW per channel is measured including the reference buffer and the sLVS serializer.

  16. Continuous Time Level Crossing Sampling ADC for Bio-Potential Recording Systems

    PubMed Central

    Tang, Wei; Osman, Ahmad; Kim, Dongsoo; Goldstein, Brian; Huang, Chenxi; Martini, Berin; Pieribone, Vincent A.

    2013-01-01

    In this paper we present a fixed window level crossing sampling analog to digital convertor for bio-potential recording sensors. This is the first proposed and fully implemented fixed window level crossing ADC without local DACs and clocks. The circuit is designed to reduce data size, power, and silicon area in future wireless neurophysiological sensor systems. We built a testing system to measure bio-potential signals and used it to evaluate the performance of the circuit. The bio-potential amplifier offers a gain of 53 dB within a bandwidth of 200 Hz-20 kHz. The input-referred rms noise is 2.8 µV. In the asynchronous level crossing ADC, the minimum delta resolution is 4 mV. The input signal frequency of the ADC is up to 5 kHz. The system was fabricated using the AMI 0.5 µm CMOS process. The chip size is 1.5 mm by 1.5 mm. The power consumption of the 4-channel system from a 3.3 V supply is 118.8 µW in the static state and 501.6 µW with a 240 kS/s sampling rate. The conversion efficiency is 1.6 nJ/conversion. PMID:24163640

  17. Development of COTS ADC SEE Test System for the ATLAS LArCalorimeter Upgrade

    DOE PAGES

    Hu, Xue -Ye; Chen, Hu -Cheng; Chen, Kai; ...

    2014-12-01

    Radiation-tolerant, high speed, high density and low power commercial off-the-shelf (COTS) analog-to-digital converters (ADCs) are planned to be used in the upgrade to the Liquid Argon (LAr) calorimeter front end (FE) trigger readout electronics. Total ionization dose (TID) and single event effect (SEE) are two important radiation effects which need to be characterized on COTS ADCs. In our initial TID test, Texas Instruments (TI) ADS5272 was identified to be the top performer after screening a total 17 COTS ADCs from different manufacturers with dynamic range and sampling rate meeting the requirements of the FE electronics. Another interesting feature of ADS5272more » is its 6.5 clock cycles latency, which is the shortest among the 17 candidates. Based on the TID performance, we have designed a SEE evaluation system for ADS5272, which allows us to further assess its radiation tolerance. In this paper, we present a detailed design of ADS5272 SEE evaluation system and show the effectiveness of this system while evaluating ADS5272 SEE characteristics in multiple irradiation tests. According to TID and SEE test results, ADS5272 was chosen to be implemented in the full-size LAr Trigger Digitizer Board (LTDB) demonstrator, which will be installed on ATLAS calorimeter during the 2014 Long Shutdown 1 (LS1).« less

  18. Development of COTS ADC SEE Test System for the ATLAS LArCalorimeter Upgrade

    SciTech Connect

    Hu, Xue -Ye; Chen, Hu -Cheng; Chen, Kai; Mead, Joseph; Liu, Shu -Bin; An, Qi

    2014-12-01

    Radiation-tolerant, high speed, high density and low power commercial off-the-shelf (COTS) analog-to-digital converters (ADCs) are planned to be used in the upgrade to the Liquid Argon (LAr) calorimeter front end (FE) trigger readout electronics. Total ionization dose (TID) and single event effect (SEE) are two important radiation effects which need to be characterized on COTS ADCs. In our initial TID test, Texas Instruments (TI) ADS5272 was identified to be the top performer after screening a total 17 COTS ADCs from different manufacturers with dynamic range and sampling rate meeting the requirements of the FE electronics. Another interesting feature of ADS5272 is its 6.5 clock cycles latency, which is the shortest among the 17 candidates. Based on the TID performance, we have designed a SEE evaluation system for ADS5272, which allows us to further assess its radiation tolerance. In this paper, we present a detailed design of ADS5272 SEE evaluation system and show the effectiveness of this system while evaluating ADS5272 SEE characteristics in multiple irradiation tests. According to TID and SEE test results, ADS5272 was chosen to be implemented in the full-size LAr Trigger Digitizer Board (LTDB) demonstrator, which will be installed on ATLAS calorimeter during the 2014 Long Shutdown 1 (LS1).

  19. Continuous Time Level Crossing Sampling ADC for Bio-Potential Recording Systems.

    PubMed

    Tang, Wei; Osman, Ahmad; Kim, Dongsoo; Goldstein, Brian; Huang, Chenxi; Martini, Berin; Pieribone, Vincent A; Culurciello, Eugenio

    2013-06-01

    In this paper we present a fixed window level crossing sampling analog to digital convertor for bio-potential recording sensors. This is the first proposed and fully implemented fixed window level crossing ADC without local DACs and clocks. The circuit is designed to reduce data size, power, and silicon area in future wireless neurophysiological sensor systems. We built a testing system to measure bio-potential signals and used it to evaluate the performance of the circuit. The bio-potential amplifier offers a gain of 53 dB within a bandwidth of 200 Hz-20 kHz. The input-referred rms noise is 2.8 µV. In the asynchronous level crossing ADC, the minimum delta resolution is 4 mV. The input signal frequency of the ADC is up to 5 kHz. The system was fabricated using the AMI 0.5 µm CMOS process. The chip size is 1.5 mm by 1.5 mm. The power consumption of the 4-channel system from a 3.3 V supply is 118.8 µW in the static state and 501.6 µW with a 240 kS/s sampling rate. The conversion efficiency is 1.6 nJ/conversion.

  20. Bethe-Salpeter study of cationic dyes: Comparisons with ADC(2) and TD-DFT

    NASA Astrophysics Data System (ADS)

    Azarias, Cloé; Duchemin, Ivan; Blase, Xavier; Jacquemin, Denis

    2017-01-01

    We present a theoretical investigation of the excited-state properties of a large series of structurally diverse arylcarbonium derivatives that are known to be challenging for theoretical models. More specifically, we compare the pros and cons of TD-DFT (TD-M06-2X), ADC(2), and BSE/GW approaches for a large panel of compounds, using two different solvent models. Both 0-0 and vertical transition energies are considered and compared to the experimental values. All approaches reasonably reproduce the auxochromic and acidochromic shifts, although in most cases both TD-DFT and BSE/GW return larger correlation with experimental values than ADC(2) for these shifts. In contrast, the absolute transition energies obtained with ADC(2) tend to be closer to the measurements, TD-DFT using the M06-2X functional largely overestimating the experimental references (by ca. 0.5 eV), and BSE/GW providing intermediate values. In addition, we show that the selected solvent model has a significant impact on the results, the corrected linear-response approach providing larger transition energies than its linear-response counterpart.

  1. Chandra and XMM Observations of the ADC Source 0921-630

    NASA Technical Reports Server (NTRS)

    Kallman, T. R.; Angelini, L.; Boroson, B.; Cottam, J.; White, Nicholas E. (Technical Monitor)

    2002-01-01

    We analyze observations of the low mass X-ray binary 2S0921-63 obtained with the gratings and CCDs on Chandra and XMM. This object is a high inclination system showing evidence for an accretion disk corona (ADC). Such a corona has the potential to constrain the properties of the heated accretion disk in this system, and other LMXBs by extension. We find evidence for line emission which is generally consistent with that found by previous experiments, although we are able to detect more lines. For the first time in this source, we find that the iron K line has multiple components. We set limits on the line widths and velocity offsets, and we fit the spectra to photoionization models and discuss the implications for accretion disk corona models. For the first time in any ADC source we use these fits, together with density constraints based on the O VII line ratio, in order to constrain the flux in the medium-ionization region of the ADC. Under various assumptions about the source luminosity this constrains the location of the emitting region. These estimates, together with estimates for the emission measure, favor a scenario in which the intrinsic luminosity of the source is comparable to what we observe.

  2. An energy-efficient and highly linear switching capacitor procedure for SAR ADCs

    NASA Astrophysics Data System (ADS)

    Rui, Ma; Wenbin, Bai; Zhangming, Zhu

    2015-05-01

    An energy-efficient and highly linear capacitor switching procedure for successive approximation register (SAR) ADCs is presented. The proposed switching procedure achieves 37% less switching energy when compared to the well-known VCM-based switching scheme. Moreover, the proposed method shows better linearity than the VCM-based one. The proposed switching procedure is applied to a 10-bit 1.0 V 300 kS/s SAR ADC implemented in 0.18 μm standard CMOS. The measured results show the SAR ADC achieves an SNDR of 55.48 dB, SFDR of 66.98 dB, and consumes 2.13 μW at a 1.0 V power supply, resulting in a figure-of-merit of 14.66 fJ/conversion-step. The measured peak DNL and INL are 0.52/-0.47 LSB and 0.72/-0.79 LSB, respectively, and the peak INL is observed at {1\\over 4}V_FS and {3\\over 4}V_FS, the same as the static nonlinearity model. Project supported by the National Natural Science Foundation of China (Nos. 61234002, 61322405, 61306044, 61376033), and the National High-Tech Program of China (Nos. 2012AA012302, 2013AA014103).

  3. Immunohistochemical evidence for the coexistence of histidine decarboxylase-like and glutamate decarboxylase-like immunoreactivities in nerve cells of the magnocellular nucleus of the posterior hypothalamus of rats.

    PubMed Central

    Takeda, N; Inagaki, S; Shiosaka, S; Taguchi, Y; Oertel, W H; Tohyama, M; Watanabe, T; Wada, H

    1984-01-01

    Immunohistochemical staining of alternate consecutive sections revealed numerous histidine decarboxylase (L-histidine carboxy-lyase, EC 4.1.1.22)-like immunoreactive neurons that also contained glutamate decarboxylase (L-glutamate 1-carboxy-lyase, EC 4.1.1.15)-like immunoreactive structures in the tuberal magnocellular nucleus, the caudal magnocellular nucleus, and the postmammillary caudal magnocellular nucleus of the posterior hypothalamus of rats. Furthermore, in immunohistochemical double-staining procedures, almost all neurons in the magnocellular nuclei had both histidine decarboxylase-like and glutamate decarboxylase-like immunoreactivities. These results suggest the coexistence of histamine and gamma-aminobutyric acid in single neurons in these nuclei. Images PMID:6594708

  4. A fast, ultra-low and frequency-scalable power consumption, 10-bit SAR ADC for particle physics detectors

    NASA Astrophysics Data System (ADS)

    Firlej, M.; Fiutowski, T.; Idzik, M.; Kulis, S.; Moron, J.; Swientek, K.

    2015-11-01

    The design and measurements results of a fast 10-bit SAR ADC with ultra-low and scalable with frequency power consumption, developed for readout systems for detectors at future particle physics colliders (ILC, CLIC, LHC Upgrade), are described. A prototype ASIC was designed and fabricated in 130 nm CMOS technology and a wide spectrum of static (INLlesssim0.5 LSB, DNLlesssim0.5 LSB) and dynamic (SINAD ~58 dB, ENOB~9.3) measurements was performed to study and quantify the ADC performance. The ADC works in wide 10 kS/s - 40 MS/s sampling frequency range, covering more than three orders of magnitude. In most of the range the power consumption scales linearly with sampling rate with a factor of about 22 μW/MS/s. A dynamic and asynchronous internal logic makes the ADC very well suited not only for commonly used synchronous sampling but also for applications with asynchronous sampling and/or the ones requiring power cycling, like the experiments at future linear collider (ILC/CLIC). The ADC layout is drawn with a small pitch of 146 μm to facilitate multi-channel integration. The obtained figure of Merit is in range 32-37 fJ/conversion for sampling frequencies 10-40 MS/s, placing the ADC among the best State of the Art designs with similar technology and specifications.

  5. Effect of the hexapeptide dalargin on ornithine decarboxylase activity in the duodenal mucosa of rats with experimental duodenal ulcer

    SciTech Connect

    Yarygin, K.N.; Shitin, A.G.; Polonskii, V.M.; Vinogradov, V.A.

    1987-08-01

    The authors study the effect of dalargin on ornithine decarboxylase in homogenates of the duodenal ulcer from rats with experimental duodenal ulcer induced by cysteamine. Activity of the enzyme was expressed in pmoles /sup 14/CO/sub 2//mg protein/h. Protein was determined by Lowry's method. The findings indicate that stimulation of ornithine decarboxylase and the antiulcerative effect of dalargin may be due to direct interaction of the peptide with cells of the intestinal mucosa and with enterocytes.

  6. Structure of ADC-68, a novel carbapenem-hydrolyzing class C extended-spectrum β-lactamase isolated from Acinetobacter baumannii.

    PubMed

    Jeon, Jeong Ho; Hong, Myoung Ki; Lee, Jung Hun; Lee, Jae Jin; Park, Kwang Seung; Karim, Asad Mustafa; Jo, Jeong Yeon; Kim, Ji Hwan; Ko, Kwan Soo; Kang, Lin Woo; Lee, Sang Hee

    2014-11-01

    Outbreaks of multidrug-resistant bacterial infections have become more frequent worldwide owing to the emergence of several different classes of β-lactamases. In this study, the molecular, biochemical and structural characteristics of an Acinetobacter-derived cephalosporinase (ADC)-type class C β-lactamase, ADC-68, isolated from the carbapenem-resistant A. baumannii D015 were investigated. The blaADC-68 gene which encodes ADC-68 was confirmed to exist on the chromosome via Southern blot analysis and draft genome sequencing. The catalytic kinetics of β-lactams and their MICs (minimum inhibitory concentrations) for A. baumannii D015 and purified ADC-68 (a carbapenemase obtained from this strain) were assessed: the strain was resistant to penicillins, narrow-spectrum and extended-spectrum cephalosporins, and carbapenems, which were hydrolyzed by ADC-68. The crystal structure of ADC-68 was determined at a resolution of 1.8 Å. The structure of ADC-68 was compared with that of ADC-1 (a non-carbapenemase); differences were found in the central part of the Ω-loop and the C-loop constituting the edge of the R1 and R2 subsites and are close to the catalytic serine residue Ser66. The ADC-68 C-loop was stabilized in the open conformation of the upper R2 subsite and could better accommodate carbapenems with larger R2 side chains. Furthermore, a wide-open conformation of the R2-loop allowed ADC-68 to bind to and hydrolyze extended-spectrum cephalosporins. Therefore, ADC-68 had enhanced catalytic efficiency against these clinically important β-lactams (extended-spectrum cephalosporins and carbapenems). ADC-68 is the first reported enzyme among the chromosomal class C β-lactamases to possess class C extended-spectrum β-lactamase and carbapenemase activities.

  7. Unusual space-group pseudo symmetry in crystals of human phosphopantothenoylcysteine decarboxylase

    SciTech Connect

    Manoj, N.; Ealick, S.E.

    2010-12-01

    Phosphopantothenoylcysteine (PPC) decarboxylase is an essential enzyme in the biosynthesis of coenzyme A and catalyzes the decarboxylation of PPC to phosphopantetheine. Human PPC decarboxylase has been expressed in Escherichia coli, purified and crystallized. The Laue class of the diffraction data appears to be {bar 3}m, suggesting space group R32 with two monomers per asymmetric unit. However, the crystals belong to the space group R3 and the asymmetric unit contains four monomers. The structure has been solved using molecular replacement and refined to a current R factor of 29%. The crystal packing can be considered as two interlaced lattices, each consistent with space group R32 and with the corresponding twofold axes parallel to each other but separated along the threefold axis. Thus, the true space group is R3 with four monomers per asymmetric unit.

  8. A coenzyme-independent decarboxylase/oxygenase cascade for the efficient synthesis of vanillin.

    PubMed

    Furuya, Toshiki; Miura, Misa; Kino, Kuniki

    2014-10-13

    Vanillin is one of the most widely used flavor compounds in the world as well as a promising versatile building block. The biotechnological production of vanillin from plant-derived ferulic acid has attracted much attention as a new alternative to chemical synthesis. One limitation of the known metabolic pathway to vanillin is its requirement for expensive coenzymes. Here, we developed a novel route to vanillin from ferulic acid that does not require any coenzymes. This artificial pathway consists of a coenzyme-independent decarboxylase and a coenzyme-independent oxygenase. When Escherichia coli cells harboring the decarboxylase/oxygenase cascade were incubated with ferulic acid, the cells efficiently synthesized vanillin (8.0 mM, 1.2 g L(-1) ) via 4-vinylguaiacol in one pot, without the generation of any detectable aromatic by-products. The efficient method described here might be applicable to the synthesis of other high-value chemicals from plant-derived aromatics.

  9. HemQ: An iron-coproporphyrin oxidative decarboxylase for protoheme synthesis in Firmicutes and Actinobacteria

    SciTech Connect

    Dailey, Harry A.; Gerdes, Svetlana

    2015-02-21

    Genes for chlorite dismutase-like proteins are found widely among heme-synthesizing bacteria and some Archaea. It is now known that among the Firmicutes and Actinobacteria these proteins do not possess chlorite dismutase activity but instead are essential for heme synthesis. These proteins, named HemQ, are ironcoproporphyrin (coproheme) decarboxylases that catalyze the oxidative decarboxylation of coproheme III into protoheme IX. As purified, HemQs do not contain bound heme, but readily bind exogeneously supplied heme with low micromolar affinity. We find that the heme-bound form of HemQ has low peroxidase activity and in the presence of peroxide the bound heme may be destroyed. Furthermore, it is possible that HemQ may serve a dual role as a decarboxylase in heme biosynthesis and a regulatory protein in heme homeostasis.

  10. HemQ: an iron-coproporphyrin oxidative decarboxylase for protoheme synthesis in Firmicutes and Actinobacteria

    PubMed Central

    Dailey, Harry A.; Gerdes, Svetlana

    2015-01-01

    Genes for chlorite dismutase-like proteins are found widely among hemesynthesizing bacteria and some Archaea. It is now known that among the Firmicutes and Actinobacteria these proteins do not possess chlorite dismutase activity but instead are essential for heme synthesis. These proteins, named HemQ, are ironcoproporphyrin (coproheme) decarboxylases that catalyze the oxidative decarboxylation of coproheme III into protoheme IX. As purified, HemQs do not contain bound heme, but readily bind exogeneously supplied heme with low micromolar affinity. The heme-bound form of HemQ has low peroxidase activity and in the presence of peroxide the bound heme may be destroyed. Thus, it is possible that HemQ may serve a dual role as a decarboxylase in heme biosynthesis and a regulatory protein in heme homeostasis. PMID:25711532

  11. Observation of Superoxide Production During Catalysis of Bacillus subtilis Oxalate Decarboxylase at pH4

    PubMed Central

    Twahir, Umar T.; Stedwell, Corey N.; Lee, Cory T.; Richards, Nigel G. J.; Polfer, Nicolas C.; Angerhofer, Alexander

    2015-01-01

    This contribution describes the trapping of the hydroperoxyl radical at a pH of 4 during turnover of wild-type oxalate decarboxylase and its T165V mutant using the spin trap BMPO. Radicals were detected and identified by a combination of EPR and mass spectrometry. Superoxide, or its conjugate acid, the hydroperoxyl radical, is expected as an intermediate in the decarboxylation and oxidation reactions of the oxalate monoanion both of which are promoted by oxalate decarboxylase. Another intermediate, the carbon dioxide radical anion was also observed. The quantitative yields of superoxide trapping is similar in the wild type and the mutant while it is significantly different for the trapping of the carbon dioxide radical anion. This suggests that the two radicals are released from different sites of the protein. PMID:25526893

  12. Increase of histidine decarboxylase activity in mice hypothalamus after intracerebroventricular administration of lipopolysaccharide.

    PubMed

    Niimi, M; Mochizuki, T; Cacabelos, R; Yamatodani, A

    1993-10-01

    The effect of intracerebroventricular (icv) administration of lipopolysaccharide on histidine decarboxylase activity and histamine content in the hypothalamus were investigated in male mice of ddY strain in vivo. Two-fold increase in histidine decarboxylase activity (HDC) was observed 4 h after administration of 50 mcg lipopolysaccharide, and HDC activity returned to the basal level within 12 h after injection. Furthermore, histamine contents showed a slight decrease at 1 and 2 h and a mild increase at 12 h after administration. However, changes in histamine content were not statistically significant. These results suggest that the increase of HDC activity in the hypothalamus by lipopolysaccharide may be involved in the central neuroimmune responses.

  13. Identification of FAH Domain-containing Protein 1 (FAHD1) as Oxaloacetate Decarboxylase*

    PubMed Central

    Pircher, Haymo; von Grafenstein, Susanne; Diener, Thomas; Metzger, Christina; Albertini, Eva; Taferner, Andrea; Unterluggauer, Hermann; Kramer, Christian; Liedl, Klaus R.; Jansen-Dürr, Pidder

    2015-01-01

    Fumarylacetoacetate hydrolase (FAH) domain-containing proteins occur in both prokaryotes and eukaryotes, where they carry out diverse enzymatic reactions, probably related to structural differences in their respective FAH domains; however, the precise relationship between structure of the FAH domain and the associated enzyme function remains elusive. In mammals, three FAH domain-containing proteins, FAHD1, FAHD2A, and FAHD2B, are known; however, their enzymatic function, if any, remains to be demonstrated. In bacteria, oxaloacetate is subject to enzymatic decarboxylation; however, oxaloacetate decarboxylases (ODx) were so far not identified in eukaryotes. Based on molecular modeling and subsequent biochemical investigations, we identified FAHD1 as a eukaryotic ODx enzyme. The results presented here indicate that dedicated oxaloacetate decarboxylases exist in eukaryotes. PMID:25575590

  14. HemQ: An iron-coproporphyrin oxidative decarboxylase for protoheme synthesis in Firmicutes and Actinobacteria

    DOE PAGES

    Dailey, Harry A.; Gerdes, Svetlana

    2015-02-21

    Genes for chlorite dismutase-like proteins are found widely among heme-synthesizing bacteria and some Archaea. It is now known that among the Firmicutes and Actinobacteria these proteins do not possess chlorite dismutase activity but instead are essential for heme synthesis. These proteins, named HemQ, are ironcoproporphyrin (coproheme) decarboxylases that catalyze the oxidative decarboxylation of coproheme III into protoheme IX. As purified, HemQs do not contain bound heme, but readily bind exogeneously supplied heme with low micromolar affinity. We find that the heme-bound form of HemQ has low peroxidase activity and in the presence of peroxide the bound heme may be destroyed.more » Furthermore, it is possible that HemQ may serve a dual role as a decarboxylase in heme biosynthesis and a regulatory protein in heme homeostasis.« less

  15. Perturbation of the Monomer-Monomer Interfaces of the Benzoylformate Decarboxylase Tetramer

    SciTech Connect

    Andrews, Forest H.; Rogers, Megan P.; Paul, Lake N.; McLeish, Michael J.

    2014-08-14

    The X-ray structure of benzoylformate decarboxylase (BFDC) from Pseudomonas putida ATCC 12633 shows it to be a tetramer. This was believed to be typical of all thiamin diphosphate-dependent decarboxylases until recently when the structure of KdcA, a branched-chain 2-keto acid decarboxylase from Lactococcus lactis, showed it to be a homodimer. This lent credence to earlier unfolding experiments on pyruvate decarboxylase from Saccharomyces cerevisiae that indicated that it might be active as a dimer. To investigate this possibility in BFDC, we sought to shift the equilibrium toward dimer formation. Point mutations were made in the noncatalytic monomer–monomer interfaces, but these had a minimal effect on both tetramer formation and catalytic activity. Subsequently, the R141E/Y288A/A306F variant was shown by analytical ultracentrifugation to be partially dimeric. It was also found to be catalytically inactive. Further experiments revealed that just two mutations, R141E and A306F, were sufficient to markedly alter the dimer–tetramer equilibrium and to provide an ~450-fold decrease in kcat. Equilibrium denaturation studies suggested that the residual activity was possibly due to the presence of residual tetramer. The structures of the R141E and A306F variants, determined to <1.5 Å resolution, hinted that disruption of the monomer interfaces will be accompanied by movement of a loop containing Leu109 and Leu110. As these residues contribute to the hydrophobicity of the active site and the correct positioning of the substrate, it seems that tetramer formation may well be critical to the catalytic activity of BFDC.

  16. Cell density-correlated induction of pyruvate decarboxylase under aerobic conditions in the yeast Pichia stipitis.

    PubMed

    Mergler, M; Klinner, U

    2001-01-01

    During the aerobic batch cultivation of P. stipitis CBS 5776 with glucose, pyruvate decarboxylase was activated in a cell number-correlated manner. Activation started when a cell number between 7 x 10(7) and x 10(8) cells ml(-1) was reached and the enzyme activity increased during further cultivation. This induction might have been triggered either by an unknown quorum sensing system or by a shortage of cytoplasmic acetyl-CoA.

  17. Arginine and lysine decarboxylases and the acid tolerance response of Salmonella Typhimurium.

    PubMed

    Alvarez-Ordóñez, Avelino; Fernández, Ana; Bernardo, Ana; López, Mercedes

    2010-01-01

    Salmonella Typhimurium CECT 443 inactivation at pH 2.5 in Mineral Medium (MM) and MM supplemented with 0.01% (w/v) arginine, lysine or glutamic acid was studied using stationary-phase cells grown in buffered BHI pH 7.0 (non-acid adapted cells) and acidified BHI up to pH 4.5 with acetic, citric, lactic and hydrochloric acids (acid adapted cells). In all cases, acid adapted cells, with D-values ranging from 23.34 to 86.90 min, showed a significantly higher acid resistance than non-acid adapted cells, with D-values between 8.90 and 10.29 min. Whereas the conditions used for acid adaptation did not exert a significant effect on the acid resistance of the S. Typhimurium CECT 443 resulting cells, the inclusion of lysine and arginine in the challenge medium protected them against acid inactivation, reaching D-values of about 2 and 3 times higher, respectively, than those found in MM or MM supplemented with glutamic acid. None of these three amino acids significantly modified the acid resistance of non-acid adapted cells. The relative expression level of adiA (encoding the arginine decarboxylase), adiY (encoding the transcriptional activator of adiA), cadA (encoding the lysine decarboxylase) and cadB (encoding the lysine/cadaverine transport protein) was examined by quantitative PCR. Acid adapted cells showed higher relative expression levels for both systems, arginine decarboxylase and lysine decarboxylase, which demonstrates that the induction of specialized pH-homeostatic systems plays an important role in S. Typhimurium CECT 443 protection against acid stress. However, the increased acid resistance showed by acid adapted cells challenged in MM arginine or lysine free suggests the existence of other microbial survival strategies.

  18. Autoradiographic measurement of relative changes in ornithine decarboxylase in axotomized superior cervical ganglion neurons

    SciTech Connect

    Wells, M.R.

    1986-05-01

    An autoradiographic method is described for detecting changes in ornithine decarboxylase in axotomized superior cervical ganglion neurons of rats using (3H)difluoromethylornithine. An increase in binding to neurons was seen at 12 h and 1 day after crushing the postganglionic nerves. Binding returned to control values between 3 and 5 days postoperation. The patterns found using this method were in general agreement with prior reports of enzymatic changes in whole ganglia.

  19. Experimental Evidence and In Silico Identification of Tryptophan Decarboxylase in Citrus Genus.

    PubMed

    De Masi, Luigi; Castaldo, Domenico; Pignone, Domenico; Servillo, Luigi; Facchiano, Angelo

    2017-02-11

    Plant tryptophan decarboxylase (TDC) converts tryptophan into tryptamine, precursor of indolealkylamine alkaloids. The recent finding of tryptamine metabolites in Citrus plants leads to hypothesize the existence of TDC activity in this genus. Here, we report for the first time that, in Citrus x limon seedlings, deuterium labeled tryptophan is decarboxylated into tryptamine, from which successively deuterated N,N,N-trimethyltryptamine is formed. These results give an evidence of the occurrence of the TDC activity and the successive methylation pathway of the tryptamine produced from the tryptophan decarboxylation. In addition, with the aim to identify the genetic basis for the presence of TDC, we carried out a sequence similarity search for TDC in the Citrus genomes using as a probe the TDC sequence reported for the plant Catharanthus roseus. We analyzed the genomes of both Citrus clementina and Citrus sinensis, available in public database, and identified putative protein sequences of aromatic l-amino acid decarboxylase. Similarly, 42 aromatic l-amino acid decarboxylase sequences from 23 plant species were extracted from public databases. Potential sequence signatures for functional TDC were then identified. With this research, we propose for the first time a putative protein sequence for TDC in the genus Citrus.

  20. Different mRNAs code for dopa decarboxylase in tissues of neuronal and nonneuronal origin

    SciTech Connect

    Krieger, M.; Coge, F.; Gros, F.; Thibault, J. )

    1991-03-15

    A cDNA clone for dopa decarboxylase has been isolated from a rat pheochromocytoma cDNA library and the cDNA sequence has been determined. It corresponds to an mRNA of 2094 nucleotides. The length of the mRNA was measured by primer-extension of rat pheochromocytoma RNA and the 5{prime} end of the sequence of the mRNA was confirmed by the PCR. A probe spanning the translation initiation site of the mRNA was used to hybridize with mRNAs from various organs of the rat. S1 nuclease digestion of the mRNAs annealed with this probe revealed two classes of mRNAs. The comparison of the cDNA sequence and published sequences for rat liver, human pheochromocytoma, and Droxophila dopa decarboxylase supported the conclusion that two mRNAs are produced: one is specific for tissue of neuronal origin and the other is specific for tissues of nonneuronal (mesodermal or endodermal) origin. The neuronal mRNA contains a 5{prime} untranslated sequence that is highly conserved between human and rat pheochromocytoma including a GA stretch. The coding sequence and the 3{prime} untranslated sequence of mRNAs from rat liver and pheochromocytoma are identical. The rat mRNA differs only in the 5{prime} untranslated region. Thus a unique gene codes for dopa decarboxylase and this gene gives rise to at least two transcripts presumably in response to different signals during development.

  1. EPR Spin Trapping of an Oxalate-Derived Free Radical in the Oxalate Decarboxylase Reaction

    PubMed Central

    Imaram, Witcha; Saylor, Benjamin T.; Centonze, Christopher P.; Richards, Nigel G. J.; Angerhofer, Alexander

    2011-01-01

    EPR spin trapping experiments on bacterial oxalate decarboxylase from Bacillus subtilis under turn-over conditions are described. The use of doubly 13C-labeled oxalate leads to a characteristic splitting of the observed radical adducts using the spin trap N-tert-butyl-α-phenylnitrone linking them directly to the substrate. The radical was identified as the carbon dioxide radical anion which is a key intermediate in the hypothetical reaction mechanism of both decarboxylase and oxidase activities. X-ray crystallography had identified a flexible loop, SENS161-4, which acts as a lid to the putative active site. Site directed mutagenesis of the hinge amino acids, S161 and T165 was explored and showed increased radical trapping yields compared to the wild type. In particular, T165V shows approximately ten times higher radical yields while at the same time its decarboxylase activity was reduced by about a factor of ten. This mutant lacks a critical H-bond between T165 and R92 resulting in compromised control over its radical chemistry allowing the radical intermediate to leak into the surrounding solution. PMID:21277974

  2. Novel protein–protein interaction between spermidine synthase and S-adenosylmethionine decarboxylase from Leishmania donovani

    SciTech Connect

    Mishra, Arjun K.; Agnihotri, Pragati; Srivastava, Vijay Kumar; Pratap, J. Venkatesh

    2015-01-09

    Highlights: • L. donovani spermidine synthase and S-adenosylmethionine decarboxylase have been cloned and purified. • S-adenosylmethionine decarboxylase has autocatalytic property. • GST pull down assay shows the two proteins to form a metabolon. • Isothermal titration calorimetry shows that binding was exothermic having K{sub d} value of 0.4 μM. • Interaction confirmed by fluorescence spectroscopy and size exclusion chromatography. - Abstract: Polyamine biosynthesis pathway has long been considered an essential drug target for trypanosomatids including Leishmania. S-adenosylmethionine decarboxylase (AdoMetDc) and spermidine synthase (SpdSyn) are enzymes of this pathway that catalyze successive steps, with the product of the former, decarboxylated S-adenosylmethionine (dcSAM), acting as an aminopropyl donor for the latter enzyme. Here we have explored the possibility of and identified the protein–protein interaction between SpdSyn and AdoMetDc. The protein–protein interaction has been identified using GST pull down assay. Isothermal titration calorimetry reveals that the interaction is thermodynamically favorable. Fluorescence spectroscopy studies also confirms the interaction, with SpdSyn exhibiting a change in tertiary structure with increasing concentrations of AdoMetDc. Size exclusion chromatography suggests the presence of the complex as a hetero-oligomer. Taken together, these results suggest that the enzymes indeed form a heteromer. Computational analyses suggest that this complex differs significantly from the corresponding human complex, implying that this complex could be a better therapeutic target than the individual enzymes.

  3. Overexpression, purification, crystallization and preliminary structural studies of p-coumaric acid decarboxylase from Lactobacillus plantarum

    SciTech Connect

    Rodríguez, Héctor; Rivas, Blanca de las; Muñoz, Rosario; Mancheño, José M.

    2007-04-01

    The enzyme p-coumaric acid decarboxylase (PDC) from L. plantarum has been recombinantly expressed, purified and crystallized. The structure has been solved at 2.04 Å resolution by the molecular-replacement method. The substrate-inducible p-coumaric acid decarboxylase (PDC) from Lactobacillus plantarum has been overexpressed in Escherichia coli, purified and confirmed to possess decarboxylase activity. The recombinant His{sub 6}-tagged enzyme was crystallized using the hanging-drop vapour-diffusion method from a solution containing 20%(w/v) PEG 4000, 12%(w/v) 2-propanol, 0.2 M sodium acetate, 0.1 M Tris–HCl pH 8.0 with 0.1 M barium chloride as an additive. Diffraction data were collected in-house to 2.04 Å resolution. Crystals belonged to the tetragonal space group P4{sub 3}, with unit-cell parameters a = b = 43.15, c = 231.86 Å. The estimated Matthews coefficient was 2.36 Å{sup 3} Da{sup −1}, corresponding to 48% solvent content, which is consistent with the presence of two protein molecules in the asymmetric unit. The structure of PDC has been determined by the molecular-replacement method. Currently, the structure of PDC complexed with substrate analogues is in progress, with the aim of elucidating the structural basis of the catalytic mechanism.

  4. 3,4-Dihydroxyphenylalanine (dopa) decarboxylase activity in the arthropod nervous system.

    PubMed

    Murdock, L L; Wirtz, R A; Köhler, G

    1973-04-01

    1. When homogenates of brains from mature adult locusts (Locusta migratoria) were incubated with l-3-(3,4-dihydroxyphenyl)[3-(14)C]alanine the major radioactive metabolite was dopamine, suggesting the presence of a dopa (3,4-dihydroxyphenylalanine) decarboxylase. 2. Decarboxylation of l-dopa by this tissue, measured under optimum conditions by a radiochemical method, was 21mumol of CO(2)/h per g wet wt. Apparent decarboxylation of l-tyrosine proceeded at 0.34mumol of CO(2)/h per g wet wt. There was no detectable decarboxylation of l-tryptophan, l-histidine or l-phenylalanine. 3. Dopa decarboxylase activity was found in all major regions of the ventral nerve cord of the mature locust (range: 4-7mumol of CO(2)/h per g wet wt.) but was low or absent in thoracic peripheral nerve. 4. Marked decarboxylation of l-dopa was found in homogenates of brains of four other species of insects, and in brain and ventral nerve cord, but not in the claw nerve, of the crayfish. 5. The activity of the locust brain enzyme may be slightly lower at the time of imaginal ecdysis than during the mature period. By contrast, the dopa decarboxylase that produces dopamine as an intermediate in cuticle biosynthesis is known to be high in activity at the time of ecdysis and low in activity during the intermoult stages.

  5. The Comparison of Wading Measurement Discharge between ADC and FP111 in Shallow Flow

    NASA Astrophysics Data System (ADS)

    Yang, H. C.; Chiang, C. J.

    2012-04-01

    During the low water period, the water depth of streams in Taiwan is very shallow and the velocity is too slow; the discharge in the streams is relatively small. At this time, larger velocity measurement equipments such as Price Type AA cannot be completely submerged in water so that they are unable to be applied to measure the velocity. Discharge measurements using conventional methods, including Price Type AA suspended in the water from bridge, are therefore unusable. The discharge records during the low water period can provide required information for monitoring ecological reference flow and various related applications of water intake engineering design. Therefore, it is necessary to choose an appropriate equipment to measure the discharge in shallow water flow in order to continuously carry out the monitoring and recording the discharge. This study is conducted in Linbian river basin, selecting Laiyi Bridge in Linbian Township Pingtung County as the gauging station. The research uses Acoustic Digital Current Meter (ADC) and Digital water velocity meter (FP111 Global Flow Probe) to measure the discharge of river during the low water period in order to explore the applicability and deviation of the two equipments which have different features and measurement principles. There are 11 discharges conducted in this study. The method of calculating the discharge is to get the sum of the measured discharge of sub-section in each measurement. Furthermore, based on velocity-area principle in Mean-Section Method, the total discharge can be estimated from the velocity data obtained by using velocity equipment. The result of the study indicates that both equipments can be applied to measure the discharge on the condition that the measurement probe should be completely submerged in water. However, ADC is more applicable than FP111 to measure discharge in shallow water flow because the precision of ADC is 7.96% higher than that of FP111. Also, through the independent sample

  6. Isolation and sequence of the gene encoding ornithine decarboxylase, SPE1, from Candida albicans by complementation of a spe1 delta strain of Saccharomyces cerevisiae.

    PubMed

    McNemar, M D; Gorman, J A; Buckley, H R

    1997-11-01

    The gene encoding ornithine decarboxylase, SPE1, from the pathogenic yeast Candida albicans has been isolated by complementation of an ornithine decarboxylase-negative (spe1 delta) strain of Saccharomyces cerevisiae. Four transformants, three of which contain plasmids with the SPE1 gene, were isolated by selection on polyamine-free medium. The C. albicans ornithine decarboxylase (ODC) showed high homology with other eukaryotic ODCs at both the amino acid and nucleic acid levels.

  7. Prognostic Value of Diffusion-Weighted Imaging (DWI) Apparent Diffusion Coefficient (ADC) in Patients with Hyperacute Cerebral Infarction Receiving rt-PA Intravenous Thrombolytic Therapy

    PubMed Central

    Sui, Hai-Jing; Yan, Cheng-Gong; Zhao, Zhen-Guo; Bai, Qing-Ke

    2016-01-01

    Background The aim of this study was to investigate the potential value of apparent diffusion coefficient (ADC) of diffusion-weighted imaging (DWI) in the prognosis of patients with hyperacute cerebral infarction (HCI) receiving intravenous thrombolytic therapy with recombinant tissue plasminogen activator (rt-PA). Material/Methods From June 2012 to June 2015, 58 cases of HCI (<6 h) undergoing rt-PA intravenous thrombolytic therapy (thrombolysis group) and 70 cases of HCI (<6 h) undergoing conventional antiplatelet and anticoagulant therapy (control group) in the same period were collected. DWI was conducted on all the subjects, and ADC maps were generated with Functool software to quantify ADC value. The clinical outcomes of HCI patients were observed for 3 months, and prognostic factors were analyzed. Results Before thrombolysis treatment, the lesion area presented high signal intensity on DWI map and low signal intensity on ADC map, and gradually weakened signal intensity on DWI map and gradually enhanced signal intensity on ADC map were observed after thrombolysis. The ADC values of the thrombolysis group were significantly higher than those of the control group after treatment (24 h, 7 d, 30 d, and 90 d) (all P<0.05), and the ADC and rADC values in the thrombolysis group gradually increased over time (all P<0.05). Multiple logistic regression analysis showed that baseline National Institutes of Health Stroke Scale (NIHSS) score, baseline rADC value, and stroke history were the independent factors for the prognosis of HIC patients with thrombolysis (all P<0.05). Conclusions The values of ADC and rADC may provide guidance in the prognosis of HCI patients receiving rt-PA, and the baseline rADC value is the protective factor for the prognosis of HCI patients receiving rt-PA. PMID:27864581

  8. Pulse oximeter improvement with an ADC-DAC feedback loop and a radial reflectance sensor.

    PubMed

    Thompson, David; Wareing, Austin; Day, Dwight; Warren, Steve

    2006-01-01

    Pulse oximeter circuitry must meet several design constraints, including the ability to separate a small pulsatile signal component from a large signal baseline. This paper describes pulse oximeter design changes that produced order-of-magnitude improvements in signal quality. The primary changes were (a) the replacement of an analog sample-and-hold-based differentiator circuit with an ADC-DAC feedback loop and (b) the replacement of a side-by-side reflectance sensor design with a radial sensor arrangement that maximizes the pulsatile-to-baseline signal ratio.

  9. 14-bit pipeline-SAR ADC for image sensor readout circuits

    NASA Astrophysics Data System (ADS)

    Wang, Gengyun; Peng, Can; Liu, Tianzhao; Ma, Cheng; Ding, Ning; Chang, Yuchun

    2015-03-01

    A two stage 14bit pipeline-SAR analog-to-digital converter includes a 5.5bit zero-crossing MDAC and a 9bit asynchronous SAR ADC for image sensor readout circuits built in 0.18um CMOS process is described with low power dissipation as well as small chip area. In this design, we employ comparators instead of high gain and high bandwidth amplifier, which consumes as low as 20mW of power to achieve the sampling rate of 40MSps and 14bit resolution.

  10. A SAR-ADC using unit bridge capacitor and with calibration for the front-end electronics of PET imaging

    NASA Astrophysics Data System (ADS)

    Liu, Wei; Wei, Tingcun; Li, Bo; Yang, Lifeng; Xue, Feifei; Hu, Yongcai

    2016-05-01

    This paper presents a 12-bit 1 MS/s successive approximation register-analog to digital converter (SAR-ADC) for the 32-channel front-end electronics of CZT-based PET imaging system. To reduce the capacitance mismatch, instead of the fractional capacitor, the unit capacitor is used as the bridge capacitor in the split-capacitor digital to analog converter (DAC) circuit. In addition, in order to eliminate the periodical DNL errors of -1 LSB which often exists in the SAR-ADC using the charge-redistributed DAC, a calibration algorithm is proposed and verified by the experiments. The proposed 12-bit 1 MS/s SAR-ADC is designed and implemented using a 0.35 μm CMOS technology, it occupies only an active area of 986×956 μm2. The measurement results show that, at the power supply of 3.3/5.0 V and the sampling rate of 1 MS/s, the ADC with calibration has a signal-to-noise-and-distortion ratio (SINAD) of 67.98 dB, the power dissipation of 5 mW, and a figure of merit (FOM) of 2.44 pJ/conv.-step. This ADC is with the features of high accuracy, low power and small layout area, it is especially suitable to the one-chip integration of the front-end readout electronics.

  11. ADC Histograms from Routine DWI for Longitudinal Studies in Cerebral Small Vessel Disease: A Field Study in CADASIL

    PubMed Central

    Gunda, Bence; Porcher, Raphael; Duering, Marco; Guichard, Jean-Pierre; Mawet, Jerome; Jouvent, Eric; Dichgans, Martin; Chabriat, Hugues

    2014-01-01

    Diffusion tensor imaging (DTI) histogram metrics are correlated with clinical parameters in cerebral small vessel diseases (cSVD). Whether ADC histogram parameters derived from simple diffusion weighted imaging (DWI) can provide relevant markers for long term studies of cSVD remains unknown. CADASIL patients were evaluated by DWI and DTI in a large cohort study overa6-year period. ADC histogram parameters were compared to those derived from mean diffusivity (MD) histograms in 280 patients using intra-class correlation and Bland-Altman plots. Impact of image corrections applied to ADC maps was assessed and a mixed effect model was used for analyzing the effects of scanner upgrades. The results showed that ADC histogram parameters are strongly correlated to MD histogram parameters and that image corrections have only limited influence on these results. Unexpectedly, scanner upgrades were found to have major effects on diffusion measures with DWI or DTI that can be even larger than those related to patients’ characteristics. These data support that ADC histograms from daily used DWI can provide relevant parameters for assessing cSVD, but the variability related to scanner upgrades as regularly performed in clinical centers should be determined precisely for longitudinal and multicentric studies using diffusion MRI in cSVD. PMID:24819368

  12. Enhancing muconic acid production from glucose and lignin-derived aromatic compounds via increased protocatechuate decarboxylase activity

    SciTech Connect

    Johnson, Christopher W.; Salvachua, Davinia; Khanna, Payal; Smith, Holly; Peterson, Darren J.; Beckham, Gregg T.

    2016-04-22

    The conversion of biomass-derived sugars and aromatic molecules to cis,cis-muconic acid (referred to hereafter as muconic acid or muconate) has been of recent interest owing to its facile conversion to adipic acid, an important commodity chemical. Metabolic routes to produce muconate from both sugars and many lignin-derived aromatic compounds require the use of a decarboxylase to convert protocatechuate (PCA, 3,4-dihydroxybenzoate) to catechol (1,2-dihydroxybenzene), two central aromatic intermediates in this pathway. Several studies have identified the PCA decarboxylase as a metabolic bottleneck, causing an accumulation of PCA that subsequently reduces muconate production. A recent study showed that activity of the PCA decarboxylase is enhanced by co-expression of two genetically associated proteins, one of which likely produces a flavin-derived cofactor utilized by the decarboxylase. Using entirely genome-integrated gene expression, we have engineered Pseudomonas putida KT2440-derived strains to produce muconate from either aromatic molecules or sugars and demonstrate in both cases that co-expression of these decarboxylase associated proteins reduces PCA accumulation and enhances muconate production relative to strains expressing the PCA decarboxylase alone. In bioreactor experiments, co-expression increased the specific productivity (mg/g cells/h) of muconate from the aromatic lignin monomer p-coumarate by 50% and resulted in a titer of >15 g/L. In strains engineered to produce muconate from glucose, co-expression more than tripled the titer, yield, productivity, and specific productivity, with the best strain producing 4.92+/-0.48 g/L muconate. Furthermore, this study demonstrates that overcoming the PCA decarboxylase bottleneck can increase muconate yields from biomass-derived sugars and aromatic molecules in industrially relevant strains and cultivation conditions.

  13. Enhancing muconic acid production from glucose and lignin-derived aromatic compounds via increased protocatechuate decarboxylase activity

    DOE PAGES

    Johnson, Christopher W.; Salvachua, Davinia; Khanna, Payal; ...

    2016-04-22

    The conversion of biomass-derived sugars and aromatic molecules to cis,cis-muconic acid (referred to hereafter as muconic acid or muconate) has been of recent interest owing to its facile conversion to adipic acid, an important commodity chemical. Metabolic routes to produce muconate from both sugars and many lignin-derived aromatic compounds require the use of a decarboxylase to convert protocatechuate (PCA, 3,4-dihydroxybenzoate) to catechol (1,2-dihydroxybenzene), two central aromatic intermediates in this pathway. Several studies have identified the PCA decarboxylase as a metabolic bottleneck, causing an accumulation of PCA that subsequently reduces muconate production. A recent study showed that activity of the PCAmore » decarboxylase is enhanced by co-expression of two genetically associated proteins, one of which likely produces a flavin-derived cofactor utilized by the decarboxylase. Using entirely genome-integrated gene expression, we have engineered Pseudomonas putida KT2440-derived strains to produce muconate from either aromatic molecules or sugars and demonstrate in both cases that co-expression of these decarboxylase associated proteins reduces PCA accumulation and enhances muconate production relative to strains expressing the PCA decarboxylase alone. In bioreactor experiments, co-expression increased the specific productivity (mg/g cells/h) of muconate from the aromatic lignin monomer p-coumarate by 50% and resulted in a titer of >15 g/L. In strains engineered to produce muconate from glucose, co-expression more than tripled the titer, yield, productivity, and specific productivity, with the best strain producing 4.92+/-0.48 g/L muconate. Furthermore, this study demonstrates that overcoming the PCA decarboxylase bottleneck can increase muconate yields from biomass-derived sugars and aromatic molecules in industrially relevant strains and cultivation conditions.« less

  14. Attitude Determination and Control Subsystem (ADCS) Preparations for the EPOXI Flyby of Comet Haley 2

    NASA Technical Reports Server (NTRS)

    Luna, Michael E.; Collins, Stephen M.

    2011-01-01

    On November 4, 2010 the already "in-flight" Deep Impact spacecraft flew within 700km of comet 103P/Hartley 2 as part of its extended mission EPOXI, the 5th time to date any spacecraft visited a comet. In 2005, the spacecraft had previously imaged a probe impact comet Tempel 1. The EPOXI flyby marked the first time in history that two comets were explored with the same instruments on a re-used spacecraft-with hardware and software originally designed and optimized for a different mission. This made the function of the attitude determination and control subsystem (ADCS) critical to the successful execution of the EPOXI flyby. As part of the spacecraft team preparations, the ADCS team had to perform thorough sequence reviews, key spacecraft activities and onboard calibrations. These activities included: review of background sequences for the initial conditions vector, sun sensor coefficients, and reaction wheel assembly (RWA) de-saturations; design and execution of 10 trajectory correction maneuvers; science calibration of the two telescope instruments; a flight demonstration of the fastest turns conducted by the spacecraft between Earth and comet point; and assessment of RWA health (given RWA problems on other spacecraft).

  15. CMOS Amperometric ADC With High Sensitivity, Dynamic Range and Power Efficiency for Air Quality Monitoring.

    PubMed

    Li, Haitao; Boling, C Sam; Mason, Andrew J

    2016-08-01

    Airborne pollutants are a leading cause of illness and mortality globally. Electrochemical gas sensors show great promise for personal air quality monitoring to address this worldwide health crisis. However, implementing miniaturized arrays of such sensors demands high performance instrumentation circuits that simultaneously meet challenging power, area, sensitivity, noise and dynamic range goals. This paper presents a new multi-channel CMOS amperometric ADC featuring pixel-level architecture for gas sensor arrays. The circuit combines digital modulation of input currents and an incremental Σ∆ ADC to achieve wide dynamic range and high sensitivity with very high power efficiency and compact size. Fabricated in 0.5 [Formula: see text] CMOS, the circuit was measured to have 164 dB cross-scale dynamic range, 100 fA sensitivity while consuming only 241 [Formula: see text] and 0.157 [Formula: see text] active area per channel. Electrochemical experiments with liquid and gas targets demonstrate the circuit's real-time response to a wide range of analyte concentrations.

  16. Relativistic decay widths of autoionization processes: The relativistic FanoADC-Stieltjes method

    NASA Astrophysics Data System (ADS)

    Fasshauer, Elke; Kolorenč, Přemysl; Pernpointner, Markus

    2015-04-01

    Electronic decay processes of ionized systems are, for example, the Auger decay or the Interatomic/ Intermolecular Coulombic Decay. In both processes, an energetically low lying vacancy is filled by an electron of an energetically higher lying orbital and a secondary electron is instantaneously emitted to the continuum. Whether or not such a process occurs depends both on the energetic accessibility and the corresponding lifetime compared to the lifetime of competing decay mechanisms. We present a realization of the non-relativistically established FanoADC-Stieltjes method for the description of autoionization decay widths including relativistic effects. This procedure, being based on the Algebraic Diagrammatic Construction (ADC), was adapted to the relativistic framework and implemented into the relativistic quantum chemistry program package Dirac. It is, in contrast to other existing relativistic atomic codes, not limited to the description of autoionization lifetimes in spherically symmetric systems, but is instead also applicable to molecules and clusters. We employ this method to the Auger processes following the Kr3d-1, Xe4d-1, and Rn5d-1 ionization. Based on the results, we show a pronounced influence of mainly scalar-relativistic effects on the decay widths of autoionization processes.

  17. Relativistic decay widths of autoionization processes: the relativistic FanoADC-Stieltjes method.

    PubMed

    Fasshauer, Elke; Kolorenč, Přemysl; Pernpointner, Markus

    2015-04-14

    Electronic decay processes of ionized systems are, for example, the Auger decay or the Interatomic/ Intermolecular Coulombic Decay. In both processes, an energetically low lying vacancy is filled by an electron of an energetically higher lying orbital and a secondary electron is instantaneously emitted to the continuum. Whether or not such a process occurs depends both on the energetic accessibility and the corresponding lifetime compared to the lifetime of competing decay mechanisms. We present a realization of the non-relativistically established FanoADC-Stieltjes method for the description of autoionization decay widths including relativistic effects. This procedure, being based on the Algebraic Diagrammatic Construction (ADC), was adapted to the relativistic framework and implemented into the relativistic quantum chemistry program package Dirac. It is, in contrast to other existing relativistic atomic codes, not limited to the description of autoionization lifetimes in spherically symmetric systems, but is instead also applicable to molecules and clusters. We employ this method to the Auger processes following the Kr3d(-1), Xe4d(-1), and Rn5d(-1) ionization. Based on the results, we show a pronounced influence of mainly scalar-relativistic effects on the decay widths of autoionization processes.

  18. A fast combination calibration of foreground and background for pipelined ADCs

    NASA Astrophysics Data System (ADS)

    Kexu, Sun; Lenian, He

    2012-06-01

    This paper describes a fast digital calibration scheme for pipelined analog-to-digital converters (ADCs). The proposed method corrects the nonlinearity caused by finite opamp gain and capacitor mismatch in multiplying digital-to-analog converters (MDACs). The considered calibration technique takes the advantages of both foreground and background calibration schemes. In this combination calibration algorithm, a novel parallel background calibration with signal-shifted correlation is proposed, and its calibration cycle is very short. The details of this technique are described in the example of a 14-bit 100 Msample/s pipelined ADC. The high convergence speed of this background calibration is achieved by three means. First, a modified 1.5-bit stage is proposed in order to allow the injection of a large pseudo-random dithering without missing code. Second, before correlating the signal, it is shifted according to the input signal so that the correlation error converges quickly. Finally, the front pipeline stages are calibrated simultaneously rather than stage by stage to reduce the calibration tracking constants. Simulation results confirm that the combination calibration has a fast startup process and a short background calibration cycle of 2 × 221 conversions.

  19. Continuous-Time ΣΔ ADC with Implicit Variable Gain Amplifier for CMOS Image Sensor

    PubMed Central

    Bermak, Amine; Abbes, Amira; Amor Benammar, Mohieddine

    2014-01-01

    This paper presents a column-parallel continuous-time sigma delta (CTSD) ADC for mega-pixel resolution CMOS image sensor (CIS). The sigma delta modulator is implemented with a 2nd order resistor/capacitor-based loop filter. The first integrator uses a conventional operational transconductance amplifier (OTA), for the concern of a high power noise rejection. The second integrator is realized with a single-ended inverter-based amplifier, instead of a standard OTA. As a result, the power consumption is reduced, without sacrificing the noise performance. Moreover, the variable gain amplifier in the traditional column-parallel read-out circuit is merged into the front-end of the CTSD modulator. By programming the input resistance, the amplitude range of the input current can be tuned with 8 scales, which is equivalent to a traditional 2-bit preamplification function without consuming extra power and chip area. The test chip prototype is fabricated using 0.18 μm CMOS process and the measurement result shows an ADC power consumption lower than 63.5 μW under 1.4 V power supply and 50 MHz clock frequency. PMID:24772012

  20. Fast Imaging Detector Readout Circuits with In-Pixel ADCs for Fourier Transform Imaging Spectrometers

    NASA Technical Reports Server (NTRS)

    Rider, D.; Blavier, J-F.; Cunningham, T.; Hancock, B.; Key, R.; Pannell, Z.; Sander, S.; Seshadri, S.; Sun, C.; Wrigley, C.

    2011-01-01

    Focal plane arrays (FPAs) with high frame rates and many pixels benefit several upcoming Earth science missions including GEO-CAPE, GACM, and ACE by enabling broader spatial coverage and higher spectral resolution. FPAs for the PanFTS, a high spatial resolution Fourier transform spectrometer and a candidate instrument for the GEO-CAPE mission are the focus of the developments reported here, but this FPA technology has the potential to enable a variety of future measurements and instruments. The ESTO ACT Program funded the developed of a fast readout integrated circuit (ROIC) based on an innovative in-pixel analog-to-digital converter (ADC). The 128 X 128 pixel ROIC features 60 ?m pixels, a 14-bit ADC in each pixel and operates at a continuous frame rate of 14 kHz consuming only 1.1 W of power. The ROIC outputs digitized data completely eliminating the bulky, power consuming signal chains needed by conventional FPAs. The 128 X 128 pixel ROIC has been fabricated in CMOS and tested at the Jet Propulsion Laboratory. The current version is designed to be hybridized with PIN photodiode arrays via indium bump bonding for light detection in the visible and ultraviolet spectral regions. However, the ROIC design incorporates a small photodiode in each cell to permit detailed characterization of the ROICperformance without the need for hybridization. We will describe the essential features of the ROIC design and present results of ROIC performance measurements.

  1. Relativistic decay widths of autoionization processes: The relativistic FanoADC-Stieltjes method

    SciTech Connect

    Fasshauer, Elke; Kolorenč, Přemysl; Pernpointner, Markus

    2015-04-14

    Electronic decay processes of ionized systems are, for example, the Auger decay or the Interatomic/ Intermolecular Coulombic Decay. In both processes, an energetically low lying vacancy is filled by an electron of an energetically higher lying orbital and a secondary electron is instantaneously emitted to the continuum. Whether or not such a process occurs depends both on the energetic accessibility and the corresponding lifetime compared to the lifetime of competing decay mechanisms. We present a realization of the non-relativistically established FanoADC-Stieltjes method for the description of autoionization decay widths including relativistic effects. This procedure, being based on the Algebraic Diagrammatic Construction (ADC), was adapted to the relativistic framework and implemented into the relativistic quantum chemistry program package Dirac. It is, in contrast to other existing relativistic atomic codes, not limited to the description of autoionization lifetimes in spherically symmetric systems, but is instead also applicable to molecules and clusters. We employ this method to the Auger processes following the Kr3d{sup −1}, Xe4d{sup −1}, and Rn5d{sup −1} ionization. Based on the results, we show a pronounced influence of mainly scalar-relativistic effects on the decay widths of autoionization processes.

  2. A 12-bit SAR ADC integrated on a multichannel silicon drift detector readout IC

    NASA Astrophysics Data System (ADS)

    Schembari, F.; Bellotti, G.; Fiorini, C.

    2016-07-01

    A 12-bit analog-to-digital converter (ADC) addressed to Silicon-Drift Detectors (SDDs) multichannel readout ASICs for X- and gamma-ray applications is presented. Aiming at digitizing output multiplexed data from the upstream analog filters banks, the converter must ensure 11-bit accuracy and a sampling frequency of about 5 MS/s. The ADC architecture is the charge-redistribution (CR) successive-approximation register (SAR). A fully differential topology has also been chosen for better rejection of common-mode noise and disturbances. The internal DAC is made of binary-scaled capacitors, whose bottom plates are switched by the SAR logic to perform the binary search of the analog input value by means of the monotonic switching scheme. The A/D converter is integrated on SFERA, a multichannel ASIC fabricated in a standard CMOS 0.35 μm 3.3 V technology and it occupies an area of 0.42 mm2. Simulated static performance shows monotonicity over the whole input-output characteristic. The description of the circuit topology and of inner blocks architectures together with the experimental characterization is here presented.

  3. A 14-bit 100-MS/s 85.2-dB SFDR pipelined ADC without calibration

    NASA Astrophysics Data System (ADS)

    Nan, Zhao; Hua, Luo; Qi, Wei; Huazhong, Yang

    2014-07-01

    This paper describes a 14-bit 100-MS/s calibration-free pipelined analog-to-digital converter (ADC). Choices for stage resolution as well as circuit topology are carefully considered to obtain high linearity without any calibration algorithm. An adjusted timing diagram with an additional clock phase is proposed to give residue voltage more settling time and minimize its distortion. The ADC employs an LVDS clock input buffer with low-jitter consideration to ensure good performance at high sampling rate. Implemented in a 0.18-μm CMOS technology, the ADC prototype achieves a spurious free dynamic range (SFDR) of 85.2 dB and signal-to-noise-and-distortion ratio (SNDR) of 63.4 dB with a 19.1-MHz input signal, while consuming 412-mW power at 2.0-V supply and occupying an area of 2.9 × 3.7mm2.

  4. A 14-bit 100-MS/s CMOS pipelined ADC featuring 83.5-dB SFDR

    NASA Astrophysics Data System (ADS)

    Nan, Zhao; Qi, Wei; Huazhong, Yang; Hui, Wang

    2014-09-01

    This paper demonstrates a 14-bit 100 MS/s CMOS pipelined analog-to-digital converter (ADC). The nonlinearity model for bootstrapped switches is established to optimize the design parameters of bootstrapped switches, and the calculations based on this model agree well with the measurement results. In order to achieve high linearity, a gradient-mismatch cancelling technique is proposed, which eliminates the first order gradient error of sampling capacitors by combining arrangement of reference control signals and capacitor layout. Fabricated in a 0.18-μm CMOS technology, this ADC occupies 10.16-mm2 area. With statistics-based background calibration of finite opamp gain in the first stage, the ADC achieves 83.5-dB spurious free dynamic range and 63.7-dB signal-to-noise-and distortion ratio respectively, and consumes 393 mW power with a supply voltage of 2 V.

  5. Low-Power Circuit Techniques for Low-Voltage Pipelined ADCs Based on Switched-Opamp Architecture

    NASA Astrophysics Data System (ADS)

    Ou, Hsin-Hung; Chang, Soon-Jyh; Liu, Bin-Da

    This paper proposes useful circuit structures for achieving a low-voltage/low-power pipelined ADC based on switched-opamp architecture. First, a novel unity-feedback-factor sample-and-hold which manipulates the features of switched-opamp technique is presented. Second, opamp-sharing is merged into switched-opamp structure with a proposed dual-output opamp configuration. A 0.8-V, 9-bit, 10-Msample/s pipelined ADC is designed to verify the proposed circuit. Simulation results using a 0.18-μm CMOS 1P6M process demonstrate the figure-of-merit of this pipelined ADC is only 0.71pJ/step.

  6. Carbon Dioxide Effects on Ethanol Production, Pyruvate Decarboxylase, and Alcohol Dehydrogenase Activities in Anaerobic Sweet Potato Roots 1

    PubMed Central

    Chang, Ling A.; Hammett, Larry K.; Pharr, David M.

    1983-01-01

    The effect of varied anaerobic atmospheres on the metabolism of sweet potato (Ipomoea batatas [L.] Lam.) roots was studied. The internal gas atmospheres of storage roots changed rapidly when the roots were submerged under water. O2 and N2 gases disappeared quickly and were replaced by CO2. There were no appreciable differences in gas composition among the four cultivars that were studied. Under different anaerobic conditions, ethanol concentration in the roots was highest in a CO2 environment, followed by submergence and a N2 environment in all the cultivars except one. A positive relationship was found between ethanol production and pyruvate decarboxylase activity from both 100% CO2-treated and 100% N2-treated roots. CO2 atmospheres also resulted in higher pyruvate decarboxylase activity than did N2 atmospheres. Concentrations of CO2 were higher within anaerobic roots than those in the ambient anaerobic atmosphere. The level of pyruvate decarboxylase and ethanol in anaerobic roots was proportional to the ambient CO2 concentration. The measurable activity of pyruvate decarboxylase that was present in the roots was about 100 times less than that of alcohol dehydrogenase. Considering these observations, it is suggested that the rate-limiting enzyme for ethanol biosynthesis in sweet potato storage roots under anoxia is likely to be pyruvate decarboxylase rather than alcohol dehydrogenase. PMID:16662798

  7. Kinetic, Mutational, and Structural Analysis of Malonate Semialdehyde Decarboxylase from Coryneform bacterium strain FG41: Mechanistic Implications for the Decarboxylase and Hydratase Activities

    PubMed Central

    Guo, Youzhong; Serrano, Hector; Poelarends, Gerrit J.; Johnson, William H.; Hackert, Marvin L.; Whitman, Christian P.

    2013-01-01

    Malonate semialdehyde decarboxylase from Pseudomonas pavonaceae 170 (designated Pp MSAD) is in a bacterial catabolic pathway for the nematicide 1,3-dichloropropene. MSAD has two known activities: it catalyzes the metal-ion independent decarboxylation of malonate semialdehyde to produce acetaldehyde and carbon dioxide, as well as a low-level hydration of 2-oxo-3-pentynoate to yield acetopyruvate. The latter activity is not known to be biologically relevant. Previous studies identified Pro-1, Asp-37, and a pair of arginines (Arg-73 and Arg-75) as critical residues in these activities. MSAD from Coryneform bacterium strain FG41 (designated FG41 MSAD) shares 38% pairwise sequence identity with the Pseudomonas enzyme including Pro-1 and Asp-37. However, Gln-73 replaces Arg-73, and the second arginine is shifted to Arg-76 by the insertion of a glycine. In order to determine how these changes relate to the activities of FG41 MSAD, the gene was cloned and the enzyme expressed and characterized. The enzyme has a comparable decarboxylase activity, but a significantly reduced hydratase activity. Mutagenesis along with crystal structures of the native enzyme (2.0 Å resolution) and the enzyme modified by a 3-oxopropanoate moiety (resulting from the incubation of enzyme and 3-bromopropiolate) (2.2 Å resolution) provided a structural basis. The roles of Pro-1 and Asp-37 are likely the same as those proposed for MSAD. However, the side chains of Thr-72, Gln-73, and Tyr-123 replace those of Arg-73 and Arg-75 in the mechanism and play a role in binding and catalysis. The structures also show that Arg-76 is likely too distant to play a direct role in the mechanism. FG41 MSAD is the second functionally annotated homologue in the MSAD family of the tautomerase superfamily and could represent a new subfamily. PMID:23781927

  8. Influence of image registration on ADC images computed from free-breathing diffusion MRIs of the abdomen

    NASA Astrophysics Data System (ADS)

    Guyader, Jean-Marie; Bernardin, Livia; Douglas, Naomi H. M.; Poot, Dirk H. J.; Niessen, Wiro J.; Klein, Stefan

    2014-03-01

    The apparent diffusion coefficient (ADC) is an imaging biomarker providing quantitative information on the diffusion of water in biological tissues. This measurement could be of relevance in oncology drug development, but it suffers from a lack of reliability. ADC images are computed by applying a voxelwise exponential fitting to multiple diffusion-weighted MR images (DW-MRIs) acquired with different diffusion gradients. In the abdomen, respiratory motion induces misalignments in the datasets, creating visible artefacts and inducing errors in the ADC maps. We propose a multistep post-acquisition motion compensation pipeline based on 3D non-rigid registrations. It corrects for motion within each image and brings all DW-MRIs to a common image space. The method is evaluated on 10 datasets of free-breathing abdominal DW-MRIs acquired from healthy volunteers. Regions of interest (ROIs) are segmented in the right part of the abdomen and measurements are compared in the three following cases: no image processing, Gaussian blurring of the raw DW-MRIs and registration. Results show that both blurring and registration improve the visual quality of ADC images, but compared to blurring, registration yields visually sharper images. Measurement uncertainty is reduced both by registration and blurring. For homogeneous ROIs, blurring and registration result in similar median ADCs, which are lower than without processing. In a ROI at the interface between liver and kidney, registration and blurring yield different median ADCs, suggesting that uncorrected motion introduces a bias. Our work indicates that averaging procedures on the scanner should be avoided, as they remove the opportunity to perform motion correction.

  9. Monitoring T2 and ADC at 9.4 T following fractionated external beam radiation therapy in a mouse model

    NASA Astrophysics Data System (ADS)

    Larocque, Matthew P.; Syme, Alasdair; Yahya, Atiyah; Wachowicz, Keith; Allalunis-Turner, Joan; Fallone, B. Gino

    2010-03-01

    The purpose of this study is to investigate the response of transverse relaxation time (T2) and apparent diffusion coefficient (ADC) in human glioma tumor xenografts during and after fractionated radiotherapy. Tumor-bearing mice were divided into four treatment groups (n = 6 per group) that received a total dose of 800 cGy of 200 kVp x-rays, given over two or three fractions, with a fraction spacing of either 24 or 72 h. A fifth treatment group received 800 cGy in a single fraction, and a sixth group of mice served as an untreated control. All mice were scanned pretreatment, before each fraction and at multiple points after treatment using a 9.4 T magnetic resonance imaging (MRI) system. Quantitative T2 and ADC maps were produced. All treated groups showed an increase in mean tumor ADC, though the time for this response to reach a maximum and return toward baseline was delayed in the fractionated groups. The highest ADC was measured 7 days after the final fraction of treatment for all groups. There were no significant differences in the maximum measured change in ADC between any of the treated groups, with the average measured maximum value being 20.5% above baseline. After treatment, all groups showed an increase in mean tumor T2, with the average measured maximum T2 being 4.7% above baseline. This increase was followed by a transition to mean T2 values below baseline values, with the average measured tumor T2 being 92.4% of the pretreatment value. The transition between elevated and depressed T2 values was delayed in the cases of fractionated therapies and occurred between 3.6 and 7.3 days after the last fraction of treatment. These results further the understanding of the temporal evolution of T2 and ADC during fractionated radiotherapy and support their potential use as time-sensitive biomarkers for tumor response.

  10. Gene cloning, recombinant expression, purification and characterization of l-methionine decarboxylase from Streptomyces sp. 590.

    PubMed

    Hayashi, Masaya; Okada, Akane; Yamamoto, Kumiko; Okugochi, Tomomi; Kusaka, Chika; Kudou, Daizou; Nemoto, Michiko; Inagaki, Junko; Hirose, Yuu; Okajima, Toshihide; Tamura, Takashi; Soda, Kenji; Inagaki, Kenji

    2016-12-21

    l-Methionine decarboxylase (MetDC) from Streptomyces sp. 590 depends on pyridoxal 5'-phosphate and catalyzes the non-oxidative decarboxylation of l-methionine to produce 3-methylthiopropylamine and carbon dioxide. MetDC gene (mdc) was determined to consist of 1,674 bp encoding 557 amino acids, and the amino acid sequence is similar to that of l-histidine decarboxylases and l-valine decarboxylases from Streptomyces sp. strains. The mdc gene was cloned and recombinant MetDC was heterologously expressed by Escherichia coli The purification of recombinant MetDC was carried out by DEAE-Toyopearl and Ni-NTA agarose column chromatography. The recombinant enzyme was homodimeric with a molecular mass of 61,000 Da and showed optimal activity between 45 to 55 °C and at pH 6.6, and the stability below 30 °C and between pH 4.6 to 7.0. l-Methionine and l-norleucine were good substrates for MetDC. The Michaelis constants for l-methionine and l-norleucine were 30 and 73 mM, respectively. The recombinant MetDC (0.50 U/ml) severely inhibited growth of human tumour cells A431 (epidermoid ovarian carcinoma cell line) and MDA-MB-231 (breast cancer cell line), however showed relatively low cytotoxicity for human normal cell NHDF-Neo (dermal fibroblast cell line from neonatal foreskin). This study revealed the properties of the gene and the protein sequence of MetDC for the first time.

  11. Biochemical Evaluation of the Decarboxylation and Decarboxylation-Deamination Activities of Plant Aromatic Amino Acid Decarboxylases*

    PubMed Central

    Torrens-Spence, Michael P.; Liu, Pingyang; Ding, Haizhen; Harich, Kim; Gillaspy, Glenda; Li, Jianyong

    2013-01-01

    Plant aromatic amino acid decarboxylase (AAAD) enzymes are capable of catalyzing either decarboxylation or decarboxylation-deamination on various combinations of aromatic amino acid substrates. These two different activities result in the production of arylalkylamines and the formation of aromatic acetaldehydes, respectively. Variations in product formation enable individual enzymes to play different physiological functions. Despite these catalytic variations, arylalkylamine and aldehyde synthesizing AAADs are indistinguishable without protein expression and characterization. In this study, extensive biochemical characterization of plant AAADs was performed to identify residues responsible for differentiating decarboxylation AAADs from aldehyde synthase AAADs. Results demonstrated that a tyrosine residue located on a catalytic loop proximal to the active site of plant AAADs is primarily responsible for dictating typical decarboxylase activity, whereas a phenylalanine at the same position is primarily liable for aldehyde synthase activity. Mutagenesis of the active site phenylalanine to tyrosine in Arabidopsis thaliana and Petroselinum crispum aromatic acetaldehyde synthases primarily converts the enzymes activity from decarboxylation-deamination to decarboxylation. The mutation of the active site tyrosine to phenylalanine in the Catharanthus roseus and Papaver somniferum aromatic amino acid decarboxylases changes the enzymes decarboxylation activity to a primarily decarboxylation-deamination activity. Generation of these mutant enzymes enables the production of unusual AAAD enzyme products including indole-3-acetaldehyde, 4-hydroxyphenylacetaldehyde, and phenylethylamine. Our data indicates that the tyrosine and phenylalanine in the catalytic loop region could serve as a signature residue to reliably distinguish plant arylalkylamine and aldehyde synthesizing AAADs. Additionally, the resulting data enables further insights into the mechanistic roles of active site

  12. Fluorimetric assay for ornithine decarboxylase by high-performance liquid chromatography.

    PubMed

    Haraguchi, K; Kai, M; Kohashi, K; Ohkura, Y

    1980-12-05

    A highly sensitive method for the assay of ornithine decarboxylase in sample solutions prepared from rat tissue homogenate is described which employs high-performance liquid chromatography with fluorescence detection. Putrescine formed from ornithine under the optimal conditions for the enzyme reaction is treated by Cellex P column chromatography for clean-up and converted into the fluorescamine derivative in the presence of cupric ion which inhibits the reaction of interfering amines with fluorescamine. The derivative is separated by reversed-phase chromatography on LiChrosorb RP-18 with linear gradient elution. The lower limit of detection for putrescine formed enzymatically is 5 pmol.

  13. Endogenous Inactivators of Arginase, l-Arginine Decarboxylase, and Agmatine Amidinohydrolase in Evernia prunastri Thallus 1

    PubMed Central

    Legaz, María Estrella; Vicente, Carlos

    1983-01-01

    Arginase (EC 3.5.3.1), l-arginine decarboxylase (EC 4.1.1.19), and agmatine amidinohydrolase (EC 3.5.3.11) activities spontaneously decay in Evernia prunastri thalli incubated on 40 millimolar l-arginine used as inducer of the three enzymes if dithiothreitol is not added to the media. Lichen thalli accumulate both chloroatranorin and evernic acid in parallel to the loss of activity. These substances behave as inactivators of the enzymes at a range of concentrations between 2 and 20 micromolar, whereas several concentrations of dithiothreitol reverse, to some extent, the in vitro inactivation. PMID:16662821

  14. Endogenous Inactivators of Arginase, l-Arginine Decarboxylase, and Agmatine Amidinohydrolase in Evernia prunastri Thallus.

    PubMed

    Legaz, M E; Vicente, C

    1983-02-01

    Arginase (EC 3.5.3.1), l-arginine decarboxylase (EC 4.1.1.19), and agmatine amidinohydrolase (EC 3.5.3.11) activities spontaneously decay in Evernia prunastri thalli incubated on 40 millimolar l-arginine used as inducer of the three enzymes if dithiothreitol is not added to the media. Lichen thalli accumulate both chloroatranorin and evernic acid in parallel to the loss of activity. These substances behave as inactivators of the enzymes at a range of concentrations between 2 and 20 micromolar, whereas several concentrations of dithiothreitol reverse, to some extent, the in vitro inactivation.

  15. Cloning of aldB, which encodes alpha-acetolactate decarboxylase, an exoenzyme from Bacillus brevis.

    PubMed Central

    Diderichsen, B; Wedsted, U; Hedegaard, L; Jensen, B R; Sjøholm, C

    1990-01-01

    A gene for alpha-acetolactate decarboxylase (ALDC) was cloned from Bacillus brevis in Escherichia coli and in Bacillus subtilis. The 1.3-kilobase-pair nucleotide sequence of the gene, aldB, encoding ALDC and its flanking regions was determined. An open reading frame of 285 amino acids included a typical N-terminal signal peptide of 24 or 27 amino acids. A B. subtilis strain harboring the aldB gene on a recombinant plasmid processed and secreted ALDC. In contrast, a similar enzyme from Enterobacter aerogenes is intracellular. Images PMID:2198252

  16. Apraxia in anti-glutamic acid decarboxylase-associated stiff person syndrome: link to corticobasal degeneration?

    PubMed

    Bowen, Lauren N; Subramony, S H; Heilman, Kenneth M

    2015-01-01

    Corticobasal syndrome (CBS) is associated with asymmetrical rigidity as well as asymmetrical limb-kinetic and ideomotor apraxia. Stiff person syndrome (SPS) is characterized by muscle stiffness and gait difficulties. Whereas patients with CBS have several forms of pathology, many patients with SPS have glutamic acid decarboxylase antibodies (GAD-ab), but these 2 disorders have not been reported to coexist. We report 2 patients with GAD-ab-positive SPS who also had signs suggestive of CBS, including asymmetrical limb rigidity associated with both asymmetrical limb-kinetic and ideomotor apraxia. Future studies should evaluate patients with CBS for GAD-ab and people with SPS for signs of CBS.

  17. Investigation of techniques to quantify in vivo lesion volume based on comparison of water apparent diffusion coefficient (ADC) maps with histology in focal cerebral ischemia of rats.

    PubMed

    Kazemi, Mark; Silva, Matthew D; Li, Fuhai; Fisher, Marc; Sotak, Christopher H

    2004-06-01

    Stroke lesion-volume estimates derived from calculated water apparent diffusion coefficient (ADC) maps provide a quantitative surrogate end-point for investigating the efficacy of drug treatment or studying the temporal evolution of cerebral ischemia. Methodology is described for estimating ischemic lesion volumes in a rat model of permanent middle cerebral artery occlusion (MCAO) based on absolute and percent-reduction threshold values of the water ADC at 3 h post-MCAO. Volume estimates derived from average ADC (ADC(av)) maps were compared with those derived from post-mortem histological sections. Optimum ADC thresholds were established as those that provided the best correlation and one-to-one correspondence between ADC- and histologically derived lesion-volume estimates. At 3 h post-MCAO, an absolute-ADC(av) threshold of 47 x 10(-5) mm(2)/s (corresponding to a 33% reduction in ADC(av) based on a contralateral hemisphere comparison) provided the most accurate estimate of percent hemispheric lesion volume (%HLV). Experimental and data analysis issues for improving and validating the usefulness of DWI as a surrogate endpoint for the quantification of ischemic lesion volume are discussed.

  18. A 1 V 186-μW 50-MS/s 10-bit subrange SAR ADC in 130-nm CMOS process

    NASA Astrophysics Data System (ADS)

    Mingyuan, Yu; Ting, Li; Jiaqi, Yang; Shuangshuang, Zhang; Fujiang, Lin; Lin, He

    2016-07-01

    This paper presents a 10-bit 50-MS/s subrange successive-approximation register (SAR) analog-to-digital converter (ADC) composed of a 4-bit SAR coarse ADC and a 6-bit SAR fine ADC. In the coarse ADC, multi-comparator SAR architecture is used to reduce the digital logic propagation delay, and a traditional asynchronous SAR ADC with monotonic switching method is used as the fine ADC. With that combination, power dissipation also can be much reduced. Meanwhile, a modified SAR control logic is adopted in the fine ADC to speed up the conversion and other techniques, such as splitting capacitors array, are borrowed to reduce the power consumption. Fabricated with 1P8M 130-nm CMOS technology, the proposed SAR ADC achieves 51.6-dB signal to noise and distortion ratio (SNDR) and consumes 186 μW at 50 MS/s with a 1-V supply, resulting in a figure of merit (FOM) of 12 fJ/conversion-step. The core area is only 0.045 mm2. Project supported by the National Natural Science Foundation of China (Nos. 61204033, 61331015), the Fundamental Research Funds for the Central Universities (No. WK2100230015), and the Funds of Science and Technology on Analog Integrated Circuit Laboratory (No. 9140C090111150C09041).

  19. Expression of Ornithine Decarboxylase Is Transiently Increased by Pollination, 2,4-Dichlorophenoxyacetic Acid, and Gibberellic Acid in Tomato Ovaries1

    PubMed Central

    Alabadí, David; Carbonell, Juan

    1998-01-01

    A cDNA encoding for a functional ornithine decarboxylase has been isolated from a cDNA library of carpels of tomato (Lycopersicon esculentum Mill.). Ornithine decarboxylase in tomato is represented by a single-copy gene that we show to be up-regulated during early fruit growth induced by 2,4-dichlorophenoxyacetic acid and gibberellic acid. PMID:9733552

  20. Amine cations promote concurrent conversion of prohistidine decarboxylase from Lactobacillus 30a to active enzyme and a modified proenzyme.

    PubMed Central

    van Poelje, P D; Snell, E E

    1988-01-01

    Activation of prohistidine decarboxylase (pi 6) from Lactobacillus 30a proceeds by an intramolecular, pH- and monovalent cation-dependent reaction in which its constituent pi chains are cleaved nonhydrolytically between Ser-81 and Ser-82 with loss of NH3 and conversion of Ser-82 to the pyruvoyl residue of active histidine decarboxylase (alpha beta)6. Amines with pKa values more than 7.0 substitute for K+ or NH4+ in the activation of prohistidine decarboxylase, but they also catalyze its inactivation in a competing reaction, pi 6----pi'6. Sequence analysis of the appropriate tryptic peptide from amine-inactivated prohistidine decarboxylase established that inactivation results from conversion of Ser-82 of the pi chain to an aminoacrylate residue. The inactivated proenzyme (pi'6) does not form histidine decarboxylase; this fact eliminates one of two postulated mechanisms of activation and, thus, favors activation by beta-elimination of the acyl group of an intermediate ester formed between Ser-81 and Ser-82. L-Histidine is bound by the proenzyme (Kd = 1.7 x 10(-4) M) and is an effective activator; one binding site is present per pi subunit. K+, NH4+, and Na+ competitively inhibit (Ki values = 2.8-4.4 x 10(-3) M) activation by histidine. The data suggest the presence of two classes of monovalent cation binding sites on prohistidine decarboxylase: one (near Ser-82) is readily saturable and one is unsaturable even by 2.4 M K+. Images PMID:3250558

  1. Local anesthetics inhibit induction of ornithine decarboxylase by the tumor promoter 12-O-tetradecanoylphorbol 13-acetate.

    PubMed Central

    Yuspa, S H; Lichti, U; Ben, T

    1980-01-01

    The induction of ornithine decarboxylase (L-ornithine carboxy-lyase, EC 4.1.1.17) activity in mouse epidermal cells in vivo and in vitro occurs rapidly after exposure to the tumor promoter 12-O-tetradecanoylphorbol 13-acetate (TPA). This induction has characteristics of a cell surface receptor-mediated process. Local anesthetics modify a variety of cellular responses mediated by membrane receptors. When cultured mouse epidermal cells were exposed to the local anesthetics lidocaine, tetracaine, or procaine (0.1-1 mM), induction of the decarboxylase by TPA was inhibited by more than 90%. In vivo, lidocaine essentially abolishes the decarboxylase response of mouse epidermis when applied shortly after TPA. In contrast, local anesthetics have no effect on the enzyme's activity when added directly to the assay mixture and, in concert with TPA, have only a minimal effect on overall protein synthesis relative to controls. However, lidocaine has no effect on TPA-stimulated DNA synthesis in vitro (12-fold with or without lidocaine). Local anesthetics also markedly inhibit induction of the decarboxylase by ultraviolet light, which is probably not membrane mediated. Furthermore, in culture, lidocaine has only a small inhibitory effect on ornithine decarboxylase when given before TPA but is an effective inhibitor even when given up to 4-5 hr after the promoter, a time when decarboxylase activity has already increased. These findings suggest that local anesthetics, which are tertiary amines, do not act at the site of interaction of TPA and its putative receptor but may be acting specifically on polyamine biosynthesis. These drugs could be useful agents to determine the role of the polyamine pathway in tumor promotion. PMID:6933562

  2. Metal Organic Framework Micro/Nanopillars of Cu(BTC)·3H2O and Zn(ADC)·DMSO

    PubMed Central

    Kojtari, Arben; Ji, Hai-Feng

    2015-01-01

    In this work, we report the optical and thermal properties of Cu(BTC)·3H2O (BTC = 1,3,5-benzenetricarboxylic acid) and Zn(ADC)·DMSO (ADC = 9,10-anthracenedicarboxylic acid, DMSO = dimethyl sulfoxide) metal-organic frameworks (MOFs) micro/nanopillars. The morphologies of MOFs on surfaces are most in the form of micro/nanopillars that were vertically oriented on the surface. The size and morphology of the pillars depend on the evaporation time, concentration, solvent, substrate, and starting volume of solutions. The crystal structures of the nanopillars and micropillars are the same, confirmed by powder XRD. Zn(ADC)·DMSO pillars have a strong blue fluorescence. Most of ADC in the pillars are in the form of monomers, which is different from ADC in the solid powder. PMID:28347026

  3. Impact of ADC non-linearities on the sensitivity to sterile keV neutrinos with a KATRIN-like experiment

    NASA Astrophysics Data System (ADS)

    Dolde, Kai; Mertens, Susanne; Radford, David; Bode, Tobias; Huber, Anton; Korzeczek, Marc; Lasserre, Thierry; Slezak, Martin

    2017-03-01

    ADC non-linearities are a major systematic effect in the search for keV-scale sterile neutrinos with tritium β-decay experiments like KATRIN. They can significantly distort the spectral shape and thereby obscure the tiny kink-like signature of a sterile neutrino. In this work we demonstrate various mitigation techniques to reduce the impact of ADC non-linearities on the tritium β-decay spectrum to a level of < ppm . The best results are achieved with a multi-pixel (≥104 pixels) detector using full waveform digitization. In this case, active-to-sterile mixing angles of the order of sin2 θ =10-7 would be accessible from the viewpoint of ADC non-linearities. With purely peak-sensing ADCs a comparable sensitivity could be reached with highly linear ADCs, sufficient non-linearity corrections or by increasing the number of pixels to ≥105 .

  4. Polyamine formation by arginine decarboxylase as a transducer of hormonal, environmental and stress stimuli in higher plants

    NASA Technical Reports Server (NTRS)

    Galston, A. W.; Flores, H. E.; Kaur-Sawhney, R.

    1982-01-01

    Recent evidence implicates polyamines including putrescine in the regulation of such diverse plant processes as cell division, embryogenesis and senescence. We find that the enzyme arginine decarboxylase, which controls the rate of putrescine formation in some plant systems, is activated by light acting through P(r) phytochrome as a receptor, by the plant hormone gibberellic acid, by osmotic shock and by other stress stimuli. We therefore propose arginine decarboxylase as a possible transducer of the various initially received tropistic stimuli in plants. The putrescine formed could act by affecting cytoskeletal components.

  5. PDC: A wire chamber cathode read-out on 6-bit fast ADC

    NASA Astrophysics Data System (ADS)

    De Giorgi, Marco; Gasparini, Fabrizio; Meneguzzo, Anna T.; Pitacco, Giorgio

    1984-06-01

    A read-out for MWPC and drift chamber is presented in which the coordinate along the sense wires is obtained by measuring the centre of gravity (CoG) of the charge induced on cathode strips or pads. The peak value of the signals coming from subsets of 8-pad cathodes are recorded by a parallel sample and hold, strobed by their own OR, and then serially digitized by one 6-bit fast ADC (FADC). The basic module of the system is peak detector and converter (PDC) built in CAMAC cards, which could be an interesting approch to the analog signal acquisition of large particle detectors. The system has been designed to equip the central detector in an experiment at the CERN LEAR facility. A prototype of a card will be described and the results of some tests will be presented.

  6. Effects of ADC Nonlinearity on the Spurious Dynamic Range Performance of Compressed Sensing

    PubMed Central

    Tian, Pengwu; Yu, Hongyi

    2014-01-01

    Analog-to-information converter (AIC) plays an important role in the compressed sensing system; it has the potential to significantly extend the capabilities of conventional analog-to-digital converter. This paper evaluates the impact of AIC nonlinearity on the dynamic performance in practical compressed sensing system, which included the nonlinearity introduced by quantization as well as the circuit non-ideality. It presents intuitive yet quantitative insights into the harmonics of quantization output of AIC, and the effect of other AIC nonlinearity on the spurious dynamic range (SFDR) performance is also analyzed. The analysis and simulation results demonstrated that, compared with conventional ADC-based system, the measurement process decorrelates the input signal and the quantization error and alleviate the effect of other decorrelates of AIC, which results in a dramatic increase in spurious free dynamic range (SFDR). PMID:24895645

  7. A PC-based single-ADC multi-parameter data acquisition system

    SciTech Connect

    Woodring, M.; Kegel, G.H.R.; Egan, J.J.

    1995-10-01

    A personal computer (PC) based mult parameter data acquisition system using the Microsoft Window operating environment has been designed and constructed. An IBI AT compatible personal computer with an Intel 486DX5 microprocessor was combined with a National Instruments ATIDIO 32 digital I/O card, a single Canberra 8713 ADC with 13-bit resolution and a modified Canberra 8223 8-input analog multiplexer to acquil data from experiments carried out at the UML Van de Graa accelerator. The accelerator data acquisition (ADAC) computer environment was programmed in Microsoft Visual BASIC for use i Windows. ADAC allows event-mode data acquisition with up to eight parameters (modifiable to 64) and the simultaneous display parameters during acquisition. Additional features of ADAC include replay of event-mode data and graphical analysis/display of data. TV ADAC environment is easy to upgrade or expand, inexpensive 1 implement, and is specifically designed to meet the needs of nuclei spectroscopy.

  8. The third-order algebraic diagrammatic construction method (ADC(3)) for the polarization propagator for closed-shell molecules: efficient implementation and benchmarking.

    PubMed

    Harbach, Philipp H P; Wormit, Michael; Dreuw, Andreas

    2014-08-14

    The implementation of an efficient program of the algebraic diagrammatic construction method for the polarisation propagator in third-order perturbation theory (ADC(3)) for the computation of excited states is reported. The accuracies of ADC(2) and ADC(3) schemes have been investigated with respect to Thiel's recently established benchmark set for excitation energies and oscillator strengths. The calculation of 141 vertical excited singlet and 71 triplet states of 28 small to medium-sized organic molecules has revealed that ADC(3) exhibits mean error and standard deviation of 0.12 ± 0.28 eV for singlet states and -0.18 ± 0.16 eV for triplet states when the provided theoretical best estimates are used as benchmark. Accordingly, the ADC(2)-s and ADC(2)-x calculations revealed accuracies of 0.22 ± 0.38 eV and -0.70 ± 0.37 eV for singlets and 0.12 ± 0.16 eV and -0.55 ± 0.20 eV for triplets, respectively. For a comparison of CC3 and ADC(3), only non-CC3 benchmark values were considered, which comprise 84 singlet states and 19 triplet states. For these singlet states CC3 exhibits an accuracy of 0.23 ± 0.21 eV and ADC(3) an accuracy of 0.08 ± 0.27 eV, and accordingly for the triplet states of 0.12 ± 0.10 eV and -0.10 ± 0.13 eV, respectively. Hence, based on the quality of the existing benchmark set it is practically not possible to judge whether ADC(3) or CC3 is more accurate, however, ADC(3) has a much larger range of applicability due to its more favourable scaling of O(N(6)) with system size.

  9. A 10 MS/s 8-bit charge-redistribution ADC for hybrid pixel applications in 65 m CMOS

    NASA Astrophysics Data System (ADS)

    Kishishita, Tetsuichi; Hemperek, Tomasz; Krüger, Hans; Koch, Manuel; Germic, Leonard; Wermes, Norbert

    2013-12-01

    The design and measurement results of an 8-bit SAR ADC, based on a charge-redistribution DAC, are presented. This ADC is characterized by superior power efficiency and small area, realized by employing a lateral metal-metal capacitor array and a dynamic two-stage comparator. To avoid the need for a high-speed clock and its associated power consumption, an asynchronous logic was implemented in a logic control cell. A test chip has been developed in a 65 nm CMOS technology, including eight ADC channels with different layout flavors of the capacitor array, a transimpedance amplifier as a signal input structure, a serializer, and a custom-made LVDS driver for data transmission. The integral (INL) and differential (DNL) nonlinearities are measured below 0.5 LSB and 0.8 LSB, respectively, for the best channel operating at a sampling frequency of 10 MS/s. The area occupies 40 μm×70 μm for one ADC channel. The power consumption is estimated as 4 μW at 1 MS/s and 38 μW at 10 MS/s with a supply rail of 1.2 V. These excellent performance features and the natural radiation hardness of the design, due to the thin gate oxide thickness of transistors, are very interesting for front-end electronics ICs of future hybrid-pixel detector systems.

  10. A 100 MS/s 9 bit 0.43 mW SAR ADC with custom capacitor array

    NASA Astrophysics Data System (ADS)

    Jingjing, Wang; Zemin, Feng; Rongjin, Xu; Chixiao, Chen; Fan, Ye; Jun, Xu; Junyan, Ren

    2016-05-01

    A low power 9 bit 100 MS/s successive approximation register analog-to-digital converter (SAR ADC) with custom capacitor array is presented. A brand-new 3-D MOM unit capacitor is used as the basic capacitor cell of this capacitor array. The unit capacitor has a capacitance of 1 fF. Besides, the advanced capacitor array structure and switch mode decrease the power consumption a lot. To verify the effectiveness of this low power design, the 9 bit 100 MS/s SAR ADC is implemented in TSMC IP9M 65 nm LP CMOS technology. The measurement results demonstrate that this design achieves an effective number of bits (ENOB) of 7.4 bit, a signal-to-noise plus distortion ratio (SNDR) of 46.40 dB and a spurious-free dynamic range (SFDR) of 62.31 dB at 100 MS/s with 1 MHz input. The SAR ADC core occupies an area of 0.030 mm2 and consumes 0.43 mW under a supply voltage of 1.2 V. The figure of merit (FOM) of the SAR ADC achieves 23.75 fJ/conv. Project supported by the National High-Tech Research and Development Program of China (No. 2013AA014101).

  11. A SHA-less 14-bit, 100-MS/s pipelined ADC with comparator offset cancellation in background

    NASA Astrophysics Data System (ADS)

    Xiaofei, Wang; Hong, Zhang; Jie, Zhang; Xin, Du; Yue, Hao

    2016-03-01

    A 14-bit 100-MS/s pipelined analog-to-digital converter (ADC) without dedicated front-end sample-and-hold amplifier (SHA) is presented. In addition to elaborate matching of the sampling network in the first stage, a background offset cancellation circuit is proposed in this paper to suppress the offset of the comparators in the first-stage sub-ADC, which ensures the overall offset does not exceed the correction range of the built-in redundant structure. Fabricated in a 0.18-μm CMOS technology, the presented ADC occupies a chip area of 12 mm2, and consumes 237 mW from a 1.8-V power supply. Measurement results with a 30.1-MHz input sine wave under a sampling rate of 100 MS/s show that the ADC achieves a 71-dB signal-to-noise and distortion ratio (SNDR), an 85.4-dB spurious-free dynamic range (SFDR), a maximum differential nonlinearity (DNL) of 0.22 LSB and a maximum integral nonlinearity (INL) of 1.4 LSB. Project supported by the National Natural Science Foundation of China (No. 61474092).

  12. A 12-bit 1 MS/s SAR-ADC for multi-channel CdZnTe detectors

    NASA Astrophysics Data System (ADS)

    Wei, Liu; Tingcun, Wei; Bo, Li; Panjie, Guo; Yongcai, Hu

    2015-04-01

    This paper presents a low power, area-efficient and radiation-hardened 12-bit 1 MS/s successive approximation register (SAR) analog-to-digital converter (ADC) for multi-channel CdZnTe (CZT) detector applications. In order to improve the SAR-ADC's accuracy, a novel comparator is proposed in which the offset voltage is self-calibrated and also a new architecture for the unit capacitor array is proposed to reduce the capacitance mismatches in the charge-redistribution DAC. The ability to radiation-harden the SAR-ADC is enhanced through circuit and layout design technologies. The prototype chip was fabricated using a TSMC 0.35 μm 2P4M CMOS process. At a 3.3/5 V power supply and a sampling rate of 1 MS/s, the proposed SAR-ADC achieves a peak signal to noise and distortion ratio (SINAD) of 67.64 dB and consumes only 10 mW power. The core of the prototype chip occupies an active area of 1180 × 1080 μm2. Project supported by the Special-Funded Program on National Key Scientific Instruments and Equipment Development (No. 2011YQ040082).

  13. A Fast Multiple Sampling Method for Low-Noise CMOS Image Sensors With Column-Parallel 12-bit SAR ADCs.

    PubMed

    Kim, Min-Kyu; Hong, Seong-Kwan; Kwon, Oh-Kyong

    2015-12-26

    This paper presents a fast multiple sampling method for low-noise CMOS image sensor (CIS) applications with column-parallel successive approximation register analog-to-digital converters (SAR ADCs). The 12-bit SAR ADC using the proposed multiple sampling method decreases the A/D conversion time by repeatedly converting a pixel output to 4-bit after the first 12-bit A/D conversion, reducing noise of the CIS by one over the square root of the number of samplings. The area of the 12-bit SAR ADC is reduced by using a 10-bit capacitor digital-to-analog converter (DAC) with four scaled reference voltages. In addition, a simple up/down counter-based digital processing logic is proposed to perform complex calculations for multiple sampling and digital correlated double sampling. To verify the proposed multiple sampling method, a 256 × 128 pixel array CIS with 12-bit SAR ADCs was fabricated using 0.18 μm CMOS process. The measurement results shows that the proposed multiple sampling method reduces each A/D conversion time from 1.2 μs to 0.45 μs and random noise from 848.3 μV to 270.4 μV, achieving a dynamic range of 68.1 dB and an SNR of 39.2 dB.

  14. A practical application of computer pattern recognition research: the Abbott ADC-500 differential classifier.

    PubMed

    Green, J E

    1979-01-01

    The ADC-500 is a new blood cell differential classifier manufactured by Abbott Laboratories. It performs 500-cell leukocyte differentials on both normal and abnormal cells, evaluates red cell morphology and estimates platelet sufficiency at a rate of 40 to 50 samples per hour in stand-alone operation. The ADC-500 system consists of a spinner which prepares a uniform blood monolayer on a slide, a stainer which reproducibly stains the slide with Wright's stain, an encoder which attaches an instrument and human readable identification to the slide and an analyzer which accepts a stack of up to 50 slides, evaluates these slides and prints the results and the slide identification on report forms. The system's analysis rate, which represents a 5- to 10-fold increase over other commercially available differential counters, requires a number of specialized techniques for its realization. One key to this performance is the development of a high speed X-Y slide positioning stage which can move to a new cell and settle in 50 msec. Another is the high degree of parallelism used in the system structure and the pipelining of the data processing. A third is the development of uniform and repeatable sample preparation modules. Within the analyzer module, the autofocus, white cell acquisition and high resolution cell analysis systems are independent and operate in parallel. At the same time within the high resolution cell analysis system, one cell is acquired; the digitized image of a second processed; and a third is classified using pattern recognition techniques. All of these tasks, except focus, are under the control of a minicomputer system. Tests of the system reveal good accuracy and an improvement in precision due to the increase in the number of counted cells.

  15. ADF/ADC Web Tools for Browsing and Visualizing Astronomical Catalogs and NASA Astrophysics Mission Metadata

    NASA Astrophysics Data System (ADS)

    Shaya, E.; Kargatis, V.; Blackwell, J.; Borne, K.; White, R. A.; Cheung, C.

    1998-05-01

    Several new web based services have been introduced this year by the Astrophysics Data Facility (ADF) at the NASA Goddard Space Flight Center. IMPReSS is a graphical interface to astrophysics databases that presents the user with the footprints of observations of space-based missions. It also aids astronomers in retrieving these data by sending requests to distributed data archives. The VIEWER is a reader of ADC astronomical catalogs and journal tables that allows subsetting of catalogs by column choices and range selection and provides database-like search capability within each table. With it, the user can easily find the table data most appropriate for their purposes and then download either the subset table or the original table. CATSEYE is a tool that plots output tables from the VIEWER (and soon AMASE), making exploring the datasets fast and easy. Having completed the basic functionality of these systems, we are enhancing the site to provide advanced functionality. These will include: market basket storage of tables and records of VIEWER output for IMPReSS and AstroBrowse queries, non-HTML table responses to AstroBrowse type queries, general column arithmetic, modularity to allow entrance into the sequence of web pages at any point, histogram plots, navigable maps, and overplotting of catalog objects on mission footprint maps. When completed, the ADF/ADC web facilities will provide astronomical tabled data and mission retrieval information in several hyperlinked environments geared for users at any level, from the school student to the typical astronomer to the expert datamining tools at state-of-the-art data centers.

  16. Molecular cloning, characterization, and function analysis of a mevalonate pyrophosphate decarboxylase gene from Ganoderma lucidum.

    PubMed

    Shi, Liang; Qin, Lei; Xu, Yingjie; Ren, Ang; Fang, Xing; Mu, Dashuai; Tan, Qi; Zhao, Mingwen

    2012-05-01

    This study investigated the role of the mevalonate pyrophosphate decarboxylase gene in the triterpene biosynthetic pathway of Ganoderma lucidum. The mevalonate pyrophosphate decarboxylase gene (mvd) was isolated using a degenerate primer-PCR technique. An analysis of the Gl-mvd transcription profile revealed a positive correlation between the expression of the Gl-mvd gene and triterpene content changes in G. lucidum during development. Furthermore, a promoter deletion analysis was conducted in G. lucidum to investigate the promoter activity and the role of methyl jasmonate (MeJA) responsive elements in the mvd promoter under the MeJA elicitor. The overexpression of Gl-mvd increased triterpene accumulation compared with the wild-type strain and increased the expression of several genes involved in the triterpene biosynthetic pathway. The findings of this study suggest that mvd may play an important role in triterpene biosynthesis regulation. Moreover, there may be the interactions among the genes involved in the triterpene biosynthetic pathway in the G. lucidum. Additionally, this study provides an approach for improving triterpene content through the overexpression of a key gene.

  17. Aromatic L-Amino Acid Decarboxylase (AADC) Is Crucial for Brain Development and Motor Functions

    PubMed Central

    Shih, De-Fen; Hsiao, Chung-Der; Min, Ming-Yuan; Lai, Wen-Sung; Yang, Chianne-Wen; Lee, Wang-Tso; Lee, Shyh-Jye

    2013-01-01

    Aromatic L-amino acid decarboxylase (AADC) deficiency is a rare pediatric neuro-metabolic disease in children. Due to the lack of an animal model, its pathogenetic mechanism is poorly understood. To study the role of AADC in brain development, a zebrafish model of AADC deficiency was generated. We identified an aadc gene homolog, dopa decarboxylase (ddc), in the zebrafish genome. Whole-mount in situ hybridization analysis showed that the ddc gene is expressed in the epiphysis, locus caeruleus, diencephalic catecholaminergic clusters, and raphe nuclei of 36-h post-fertilization (hpf) zebrafish embryos. Inhibition of Ddc by AADC inhibitor NSD-1015 or anti-sense morpholino oligonucleotides (MO) reduced brain volume and body length. We observed increased brain cell apoptosis and loss of dipencephalic catecholaminergic cluster neurons in ddc morphants (ddc MO-injected embryos). Seizure-like activity was also detected in ddc morphants in a dose-dependent manner. ddc morphants had less sensitive touch response and impaired swimming activity that could be rescued by injection of ddc plasmids. In addition, eye movement was also significantly impaired in ddc morphants. Collectively, loss of Ddc appears to result in similar phenotypes as that of ADCC deficiency, thus zebrafish could be a good model for investigating pathogenetic mechanisms of AADC deficiency in children. PMID:23940784

  18. Production of pyruvate from mannitol by mannitol-assimilating pyruvate decarboxylase-negative Saccharomyces cerevisiae

    PubMed Central

    Yoshida, Shiori; Tanaka, Hideki; Hirayama, Makoto; Murata, Kousaku; Kawai, Shigeyuki

    2015-01-01

    Mannitol is contained in brown macroalgae up to 33% (w/w, dry weight), and thus is a promising carbon source for white biotechnology. However, Saccharomyces cerevisiae, a key cell factory, is generally regarded to be unable to assimilate mannitol for growth. We have recently succeeded in producing S. cerevisiae that can assimilate mannitol through spontaneous mutations of Tup1-Cyc8, each of which constitutes a general corepressor complex. In this study, we demonstrate production of pyruvate from mannitol using this mannitol-assimilating S. cerevisiae through deletions of all 3 pyruvate decarboxylase genes. The resultant mannitol-assimilating pyruvate decarboxylase-negative strain produced 0.86 g/L pyruvate without use of acetate after cultivation for 4 days, with an overall yield of 0.77 g of pyruvate per g of mannitol (the theoretical yield was 79%). Although acetate was not needed for growth of this strain in mannitol-containing medium, addition of acetate had a significant beneficial effect on production of pyruvate. This is the first report of production of a valuable compound (other than ethanol) from mannitol using S. cerevisiae, and is an initial platform from which the productivity of pyruvate from mannitol can be improved. PMID:26588105

  19. Crystal structure of Mycobacterium tuberculosis diaminopimelate decarboxylase, an essential enzyme in bacterial lysine biosynthesis.

    PubMed

    Gokulan, Kuppan; Rupp, Bernhard; Pavelka, Martin S; Jacobs, William R; Sacchettini, James C

    2003-05-16

    The Mycobacterium tuberculosis lysA gene encodes the enzyme meso-diaminopimelate decarboxylase (DAPDC), a pyridoxal-5'-phosphate (PLP)-dependent enzyme. The enzyme catalyzes the final step in the lysine biosynthetic pathway converting meso-diaminopimelic acid (DAP) to l-lysine. The lysA gene of M. tuberculosis H37Rv has been established as essential for bacterial survival in immunocompromised mice, demonstrating that de novo biosynthesis of lysine is essential for in vivo viability. Drugs targeted against DAPDC could be efficient anti-tuberculosis drugs, and the three-dimensional structure of DAPDC from M. tuberculosis complexed with reaction product lysine and the ternary complex with PLP and lysine in the active site has been determined. The first structure of a DAPDC confirms its classification as a fold type III PLP-dependent enzyme. The structure shows a stable 2-fold dimer in head-to-tail arrangement of a triose-phosphate isomerase (TIM) barrel-like alpha/beta domain and a C-terminal beta sheet domain, similar to the ornithine decarboxylase (ODC) fold family. PLP is covalently bound via an internal aldimine, and residues from both domains and both subunits contribute to the binding pocket. Comparison of the structure with eukaryotic ODCs, in particular with a di-fluoromethyl ornithine (DMFO)-bound ODC from Trypanosoma bruceii, indicates that corresponding DAP-analogues might be potential inhibitors for mycobacterial DAPDCs.

  20. Production of pyruvate from mannitol by mannitol-assimilating pyruvate decarboxylase-negative Saccharomyces cerevisiae.

    PubMed

    Yoshida, Shiori; Tanaka, Hideki; Hirayama, Makoto; Murata, Kousaku; Kawai, Shigeyuki

    2015-01-01

    Mannitol is contained in brown macroalgae up to 33% (w/w, dry weight), and thus is a promising carbon source for white biotechnology. However, Saccharomyces cerevisiae, a key cell factory, is generally regarded to be unable to assimilate mannitol for growth. We have recently succeeded in producing S. cerevisiae that can assimilate mannitol through spontaneous mutations of Tup1-Cyc8, each of which constitutes a general corepressor complex. In this study, we demonstrate production of pyruvate from mannitol using this mannitol-assimilating S. cerevisiae through deletions of all 3 pyruvate decarboxylase genes. The resultant mannitol-assimilating pyruvate decarboxylase-negative strain produced 0.86 g/L pyruvate without use of acetate after cultivation for 4 days, with an overall yield of 0.77 g of pyruvate per g of mannitol (the theoretical yield was 79%). Although acetate was not needed for growth of this strain in mannitol-containing medium, addition of acetate had a significant beneficial effect on production of pyruvate. This is the first report of production of a valuable compound (other than ethanol) from mannitol using S. cerevisiae, and is an initial platform from which the productivity of pyruvate from mannitol can be improved.

  1. Hypothalamic L-Histidine Decarboxylase Is Up-Regulated During Chronic REM Sleep Deprivation of Rats

    PubMed Central

    Hoffman, Gloria E.; Koban, Michael

    2016-01-01

    A competition of neurobehavioral drives of sleep and wakefulness occurs during sleep deprivation. When enforced chronically, subjects must remain awake. This study examines histaminergic neurons of the tuberomammillary nucleus of the posterior hypothalamus in response to enforced wakefulness in rats. We tested the hypothesis that the rate-limiting enzyme for histamine biosynthesis, L-histidine decarboxylase (HDC), would be up-regulated during chronic rapid eye movement sleep deprivation (REM-SD) because histamine plays a major role in maintaining wakefulness. Archived brain tissues of male Sprague Dawley rats from a previous study were used. Rats had been subjected to REM-SD by the flowerpot paradigm for 5, 10, or 15 days. For immunocytochemistry, rats were transcardially perfused with acrolein-paraformaldehyde for immunodetection of L-HDC; separate controls used carbodiimide-paraformaldehyde for immunodetection of histamine. Immunolocalization of histamine within the tuberomammillary nucleus was validated using carbodiimide. Because HDC antiserum has cross-reactivity with other decarboxylases at high antibody concentrations, titrations localized L-HDC to only tuberomammillary nucleus at a dilution of ≥ 1:300,000. REM-SD increased immunoreactive HDC by day 5 and it remained elevated in both dorsal and ventral aspects of the tuberomammillary complex. Our results suggest that up-regulation of L-HDC within the tuberomammillary complex during chronic REM-SD may be responsible for maintaining wakefulness. PMID:27997552

  2. Rational design of ornithine decarboxylase with high catalytic activity for the production of putrescine.

    PubMed

    Choi, Hyang; Kyeong, Hyun-Ho; Choi, Jung Min; Kim, Hak-Sung

    2014-09-01

    Putrescine finds wide industrial applications in the synthesis of polymers, pharmaceuticals, agrochemicals, and surfactants. Owing to economic and environmental concerns, the microbial production of putrescine has attracted a great deal of attention, and ornithine decarboxylase (ODC) is known to be a key enzyme in the biosynthetic pathway. Herein, we present the design of ODC from Escherichia coli with high catalytic efficiency using a structure-based rational approach. Through a substrate docking into the model structure of the enzyme, we first selected residues that might lead to an increase in catalytic activity. Of the selected residues that are located in the α-helix and the loops constituting the substrate entry site, a mutational analysis of the single mutants identified two key residues, I163 and E165. A combination of two single mutations resulted in a 62.5-fold increase in the catalytic efficiency when compared with the wild-type enzyme. Molecular dynamics simulations of the best mutant revealed that the substrate entry site becomes more flexible through mutations, while stabilizing the formation of the dimeric interface of the enzyme. Our approach can be applied to the design of other decarboxylases with high catalytic efficiency for the production of various chemicals through bio-based processes.

  3. Decarboxylase gene expression and cadaverine and putrescine production by Serratia proteamaculans in vitro and in beef.

    PubMed

    De Filippis, Francesca; Pennacchia, Carmela; Di Pasqua, Rosangela; Fiore, Alberto; Fogliano, Vincenzo; Villani, Francesco; Ercolini, Danilo

    2013-08-01

    Studies of the molecular basis of microbial metabolic activities that are important for the changes in food quality are valuable in order to help in understanding the behavior of spoiling bacteria in food. The growth of a psychrotrophic Serratia proteamaculans strain was monitored in vitro and in artificially inoculated raw beef. Two growth temperatures (25°C and 4°C) were tested in vitro, while growth at 15°C and 4°C was monitored in beef. During growth, the expression of inducible lysine and ornithine-decarboxylase genes was evaluated by quantitative reverse transcription-PCR (qRT-PCR), while the presence of cadaverine and putrescine was quantified by LC-ESI-MS/MS. The expression of the decarboxylase genes, and the consequent production of cadaverine and putrescine were shown to be influenced by the temperature, as well as by the complexity of the growth medium. Generally, the maximum gene expression and amine production took place during the exponential and early stationary phase, respectively. In addition, lower temperatures caused slower growth and gene downregulation. Higher amounts of cadaverine compared to putrescine were found during growth in beef with the highest concentrations corresponding to microbial loads of ca. 9CFU/g. The differences found in gene expression evaluated in vitro and in beef suggested that such activities are more reliably investigated in situ in specific food matrices.

  4. NUTRITIONAL FACTORS STIMULATING THE FORMATION OF LYSINE DECARBOXYLASE IN ESCHERICHIA COLI

    PubMed Central

    Maretzki, Andrew; Mallette, M. F.

    1962-01-01

    Maretzki, Andrew (Pennsylvania State University, University Park) and M. F. Mallette. Nutritional factors stimulating the formation of lysine decarboxylase in Escherichia coli. J. Bacteriol. 83:720–726. 1962 — Inclusion of complex nitrogen sources in the induction medium was shown to be necessary for the synthesis of appreciable amounts of l-lysine decarboxylase by Escherichia coli B. Hy-case, a commercial acid hydrolyzate of casein, was especially effective in enzyme production, which was assayed manometrically after lysis of the bacteria from without by bacteriophage. Partial fractionation of the Hy-case, identification of the free amino acids, and addition of these amino acids to test media revealed stimulatory effects by methionine, threonine, proline, leucine, and tyrosine. A full complement of amino acids did not match the enzyme levels reached in the presence of Hy-case. Certain peptide fractions obtained from this mixture supplemented the effects of the amino acids in such a way as to suggest direct incorporation of peptide rather than transport or protective roles. Added purines, pyrimidines, iron, and water-soluble vitamins were without effect. Neither carbohydrates nor phosphorylated materials could be detected in the stimulatory fractions. PMID:14469751

  5. Crystal Structure and Substrate Specificity of Drosophila 3,4-Dihydroxyphenylalanine Decarboxylase

    SciTech Connect

    Han, Q.; Ding, H; Robinson, H; Christensen, B; Li, J

    2010-01-01

    3,4-Dihydroxyphenylalanine decarboxylase (DDC), also known as aromatic L-amino acid decarboxylase, catalyzes the decarboxylation of a number of aromatic L-amino acids. Physiologically, DDC is responsible for the production of dopamine and serotonin through the decarboxylation of 3,4-dihydroxyphenylalanine and 5-hydroxytryptophan, respectively. In insects, both dopamine and serotonin serve as classical neurotransmitters, neuromodulators, or neurohormones, and dopamine is also involved in insect cuticle formation, eggshell hardening, and immune responses. In this study, we expressed a typical DDC enzyme from Drosophila melanogaster, critically analyzed its substrate specificity and biochemical properties, determined its crystal structure at 1.75 Angstrom resolution, and evaluated the roles residues T82 and H192 play in substrate binding and enzyme catalysis through site-directed mutagenesis of the enzyme. Our results establish that this DDC functions exclusively on the production of dopamine and serotonin, with no activity to tyrosine or tryptophan and catalyzes the formation of serotonin more efficiently than dopamine. The crystal structure of Drosophila DDC and the site-directed mutagenesis study of the enzyme demonstrate that T82 is involved in substrate binding and that H192 is used not only for substrate interaction, but for cofactor binding of drDDC as well. Through comparative analysis, the results also provide insight into the structure-function relationship of other insect DDC-like proteins.

  6. Characterization of Plasmodium phosphatidylserine decarboxylase expressed in yeast and application for inhibitor screening.

    PubMed

    Choi, Jae-Yeon; Kumar, Vidya; Pachikara, Niseema; Garg, Aprajita; Lawres, Lauren; Toh, Justin Y; Voelker, Dennis R; Ben Mamoun, Choukri

    2016-03-01

    Phospholipid biosynthesis is critical for the development, differentiation and pathogenesis of several eukaryotic pathogens. Genetic studies have validated the pathway for phosphatidylethanolamine synthesis from phosphatidylserine catalyzed by phosphatidylserine decarboxylase enzymes (PSD) as a suitable target for development of antimicrobials; however no inhibitors of this class of enzymes have been discovered. We show that the Plasmodium falciparum PSD can restore the essential function of the yeast gene in strains requiring PSD for growth. Genetic, biochemical and metabolic analyses demonstrate that amino acids between positions 40 and 70 of the parasite enzyme are critical for proenzyme processing and decarboxylase activity. We used the essential role of Plasmodium PSD in yeast as a tool for screening a library of anti-malarials. One of these compounds is 7-chloro-N-(4-ethoxyphenyl)-4-quinolinamine, an inhibitor with potent activity against P. falciparum, and low toxicity toward mammalian cells. We synthesized an analog of this compound and showed that it inhibits PfPSD activity and eliminates Plasmodium yoelii infection in mice. These results highlight the importance of 4-quinolinamines as a novel class of drugs targeting membrane biogenesis via inhibition of PSD activity.

  7. Transport of phosphatidylserine from the endoplasmic reticulum to the site of phosphatidylserine decarboxylase2 in yeast.

    PubMed

    Kannan, Muthukumar; Riekhof, Wayne R; Voelker, Dennis R

    2015-02-01

    Over the past two decades, most of the genes specifying lipid synthesis and metabolism in yeast have been identified and characterized. Several of these biosynthetic genes and their encoded enzymes have provided valuable tools for the genetic and biochemical dissection of interorganelle lipid transport processes in yeast. One such pathway involves the synthesis of phosphatidylserine (PtdSer) in the endoplasmic reticulum (ER), and its non-vesicular transport to the site of phosphatidylserine decarboxylase2 (Psd2p) in membranes of the Golgi and endosomal sorting system. In this review, we summarize the identification and characterization of the yeast phosphatidylserine decarboxylases, and examine their role in studies of the transport-dependent pathways of de novo synthesis of phosphatidylethanolamine (PtdEtn). The emerging picture of the Psd2p-specific transport pathway is one in which the enzyme and its non-catalytic N-terminal domains act as a hub to nucleate the assembly of a multiprotein complex, which facilitates PtdSer transport at membrane contact sites between the ER and Golgi/endosome membranes. After transport to the catalytic site of Psd2p, PtdSer is decarboxylated to form PtdEtn, which is disseminated throughout the cell to support the structural and functional needs of multiple membranes.

  8. Environmental stress causes oxidative damage to plant mitochondria leading to inhibition of glycine decarboxylase.

    PubMed

    Taylor, Nicolas L; Day, David A; Millar, A Harvey

    2002-11-08

    A cytotoxic product of lipid peroxidation, 4-hydroxy-2-nonenal (HNE), rapidly inhibited glycine, malate/pyruvate, and 2-oxoglutarate-dependent O2 consumption by pea leaf mitochondria. Dose- and time-dependence of inhibition showed that glycine oxidation was the most severely affected with a K(0.5) of 30 microm. Several mitochondrial proteins containing lipoic acid moieties differentially lost their reactivity to a lipoic acid antibody following HNE treatment. The most dramatic loss of antigenicity was seen with the 17-kDa glycine decarboxylase complex (GDC) H-protein, which was correlated with the loss of glycine-dependent O2 consumption. Paraquat treatment of pea seedlings induced lipid peroxidation, which resulted in the rapid loss of glycine-dependent respiration and loss of H-protein reactivity with lipoic acid antibodies. Pea plants exposed to chilling and water deficit responded similarly. In contrast, the damage to other lipoic acid-containing mitochondrial enzymes was minor under these conditions. The implication of the acute sensitivity of glycine decarboxylase complex H-protein to lipid peroxidation products is discussed in the context of photorespiration and potential repair mechanisms in plant mitochondria.

  9. Overexpression, purification, crystallization and preliminary structural studies of p-coumaric acid decarboxylase from Lactobacillus plantarum

    PubMed Central

    Rodríguez, Héctor; de las Rivas, Blanca; Muñoz, Rosario; Mancheño, José M.

    2007-01-01

    The substrate-inducible p-coumaric acid decarboxylase (PDC) from Lactobacillus plantarum has been overexpressed in Escherichia coli, purified and confirmed to possess decarboxylase activity. The recombinant His6-tagged enzyme was crystallized using the hanging-drop vapour-diffusion method from a solution containing 20%(w/v) PEG 4000, 12%(w/v) 2-propanol, 0.2 M sodium acetate, 0.1 M Tris–HCl pH 8.0 with 0.1 M barium chloride as an additive. Diffraction data were collected in-house to 2.04 Å resolution. Crystals belonged to the tetragonal space group P43, with unit-cell parameters a = b = 43.15, c = 231.86 Å. The estimated Matthews coefficient was 2.36 Å3 Da−1, corresponding to 48% solvent content, which is consistent with the presence of two protein molecules in the asymmetric unit. The structure of PDC has been determined by the molecular-replacement method. Currently, the structure of PDC complexed with substrate analogues is in progress, with the aim of elucidating the structural basis of the catalytic mechanism. PMID:17401200

  10. Evolutionary Trails of Plant Group II Pyridoxal Phosphate-Dependent Decarboxylase Genes

    PubMed Central

    Kumar, Rahul

    2016-01-01

    Type II pyridoxal phosphate-dependent decarboxylase (PLP_deC) enzymes play important metabolic roles during nitrogen metabolism. Recent evolutionary profiling of these genes revealed a sharp expansion of histidine decarboxylase genes in the members of Solanaceae family. In spite of the high sequence homology shared by PLP_deC orthologs, these enzymes display remarkable differences in their substrate specificities. Currently, limited information is available on the gene repertoires and substrate specificities of PLP_deCs which renders their precise annotation challenging and offers technical challenges in the immediate identification and biochemical characterization of their full gene complements in plants. Herein, we explored their evolutionary trails in a comprehensive manner by taking advantage of high-throughput data accessibility and computational approaches. We discussed the premise that has enabled an improved reconstruction of their evolutionary lineage and evaluated the factors offering constraints in their rapid functional characterization, till date. We envisage that the synthesized information herein would act as a catalyst for the rapid exploration of their biochemical specificity and physiological roles in more plant species. PMID:27602045

  11. Characterization of Plasmodium phosphatidylserine decarboxylase expressed in yeast and application for inhibitor screening

    PubMed Central

    Choi, Jae-Yeon; Lawres, Lauren; Toh, Justin Y.; Voelker, Dennis R.; Ben Mamoun, Choukri

    2016-01-01

    Summary Phospholipid biosynthesis is critical for the development, differentiation and pathogenesis of several eukaryotic pathogens. Genetic studies have validated the pathway for phosphatidylethanolamine synthesis from phosphatidylserine catalyzed by phosphatidylserine decarboxylase enzymes (PSD) as a suitable target for development of antimicrobials; however no inhibitors of this class of enzymes have been discovered. We show that the Plasmodium falciparum PSD can restore the essential function of the yeast gene in strains requiring PSD for growth. Genetic, biochemical and metabolic analyses demonstrate that amino acids between positions 40 and 70 of the parasite enzyme are critical for proenzyme processing and decarboxylase activity. We used the essential role of Plasmodium PSD in yeast as a tool for screening a library of anti-malarials. One of these compounds is 7-chloro-N-(4-ethoxyphenyl)-4-quinolinamine, an inhibitor with potent activity against P. falciparum, and low toxicity toward mammalian cells. We synthesized an analog of this compound and showed that it inhibits PfPSD activity and eliminates Plasmodium yoelii infection in mice. These results highlight the importance of 4-quinolinamines as a novel class of drugs targeting membrane biogenesis via inhibition of PSD activity PMID:26585333

  12. Hypothalamic L-Histidine Decarboxylase Is Up-Regulated During Chronic REM Sleep Deprivation of Rats.

    PubMed

    Hoffman, Gloria E; Koban, Michael

    2016-01-01

    A competition of neurobehavioral drives of sleep and wakefulness occurs during sleep deprivation. When enforced chronically, subjects must remain awake. This study examines histaminergic neurons of the tuberomammillary nucleus of the posterior hypothalamus in response to enforced wakefulness in rats. We tested the hypothesis that the rate-limiting enzyme for histamine biosynthesis, L-histidine decarboxylase (HDC), would be up-regulated during chronic rapid eye movement sleep deprivation (REM-SD) because histamine plays a major role in maintaining wakefulness. Archived brain tissues of male Sprague Dawley rats from a previous study were used. Rats had been subjected to REM-SD by the flowerpot paradigm for 5, 10, or 15 days. For immunocytochemistry, rats were transcardially perfused with acrolein-paraformaldehyde for immunodetection of L-HDC; separate controls used carbodiimide-paraformaldehyde for immunodetection of histamine. Immunolocalization of histamine within the tuberomammillary nucleus was validated using carbodiimide. Because HDC antiserum has cross-reactivity with other decarboxylases at high antibody concentrations, titrations localized L-HDC to only tuberomammillary nucleus at a dilution of ≥ 1:300,000. REM-SD increased immunoreactive HDC by day 5 and it remained elevated in both dorsal and ventral aspects of the tuberomammillary complex. Our results suggest that up-regulation of L-HDC within the tuberomammillary complex during chronic REM-SD may be responsible for maintaining wakefulness.

  13. Absence of malonyl coenzyme A decarboxylase in mice increases cardiac glucose oxidation and protects the heart from ischemic injury

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Acute pharmacological inhibition of cardiac malonyl coenzyme A decarboxylase (MCD) protects the heart from ischemic damage by inhibiting fatty acid oxidation and stimulating glucose oxidation. However, it is unknown whether chronic inhibition of MCD results in altered cardiac function, energy metabo...

  14. 21 CFR 173.115 - Alpha-acetolactate decarboxylase (α-ALDC) enzyme preparation derived from a recombinant Bacillus...

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... preparation derived from a recombinant Bacillus subtilis. 173.115 Section 173.115 Food and Drugs FOOD AND DRUG... Bacillus subtilis. The food additive alpha-acetolactate decarboxylase (α-ALDC) enzyme preparation, may be... derived from a modified Bacillus subtilis strain that contains the gene coding for α-ALDC from...

  15. Molecular analysis of a new member of the opium poppy tyrosine/3,4-dihydroxyphenylalanine decarboxylase gene family.

    PubMed Central

    Maldonado-Mendoza, I E; López-Meyer, M; Galef, J R; Burnett, R J; Nessler, C L

    1996-01-01

    An aromatic amino acid decarboxylase DNA fragment was generated from opium poppy (Papaver somniferum L.) genomic DNA by the PCR using primers designed from conserved amino acid sequences of other aromatic amino acid decarboxylase genes. Using this fragment as a probe, a genomic clone was isolated that encodes a new member of the opium poppy tyrosine/3,4-dihydroxyphenylalanine decarboxylase gene family (TyDC5). The predicted TyDC5 amino acid sequence shares extensive identity with other opium poppy tyrosine/3,4-dihydroxyphenylalanine decarboxylases (84%), and when expressed in Escherichia coli, it is active against tyrosine and to a lesser extent against 3,4-dihydroxyphenylalanine. Ribonuclease protection assays indicate that TyDC5 is expressed primarily in the roots of mature poppy plants. A peak of TyDC5 expression was also observed during germination, coincident with the emergence of the radicle from the seed coat. Parallel results were obtained in transgenic tobacco using a TyDC5 promoter fragment (-2060) translationally fused to the beta-glucuronidase reporter gene (GUS). IN TyDC5::GUS tobacco, GUS activity transiently appeared in all parts of the seedling during germination, but was limited to the roots in older plants. These results indicate that TyDC5 expression is transcriptionally regulated and suggest that the TyDC5 enzyme may play an important role in providing precursors for alkaloid synthesis in the roots and germinating seedlings of opium poppy. PMID:8587993

  16. CONFIRMATIONAL IDENTIFICATION OF ESCHERICHIA COLI, A COMPARISON OF GENOTYPIC AND PHENOTYPIC ASSAYS FOR GLUTAMATE DECARBOXYLASE AND B-D-GLUCURONIDASE

    EPA Science Inventory

    Genotypic and phenotypic assays for glutamate decarboxylase (GAD) and B-D-glucuronidase (GUD) were compared for their abilities to detect various strains of Escherichia coli and to discriminate among other bacterial species. Test strains included nonpathogenic E.coli, three major...

  17. In vitro Characterization of Phenylacetate Decarboxylase, a Novel Enzyme Catalyzing Toluene Biosynthesis in an Anaerobic Microbial Community

    PubMed Central

    Zargar, K.; Saville, R.; Phelan, R. M.; Tringe, S. G.; Petzold, C. J.; Keasling, J. D.; Beller, H. R.

    2016-01-01

    Anaerobic bacterial biosynthesis of toluene from phenylacetate was reported more than two decades ago, but the biochemistry underlying this novel metabolism has never been elucidated. Here we report results of in vitro characterization studies of a novel phenylacetate decarboxylase from an anaerobic, sewage-derived enrichment culture that quantitatively produces toluene from phenylacetate; complementary metagenomic and metaproteomic analyses are also presented. Among the noteworthy findings is that this enzyme is not the well-characterized clostridial p-hydroxyphenylacetate decarboxylase (CsdBC). However, the toluene synthase under study appears to be able to catalyze both phenylacetate and p-hydroxyphenylacetate decarboxylation. Observations suggesting that phenylacetate and p-hydroxyphenylacetate decarboxylation in complex cell-free extracts were catalyzed by the same enzyme include the following: (i) the specific activity for both substrates was comparable in cell-free extracts, (ii) the two activities displayed identical behavior during chromatographic separation of cell-free extracts, (iii) both activities were irreversibly inactivated upon exposure to O2, and (iv) both activities were similarly inhibited by an amide analog of p-hydroxyphenylacetate. Based upon these and other data, we hypothesize that the toluene synthase reaction involves a glycyl radical decarboxylase. This first-time study of the phenylacetate decarboxylase reaction constitutes an important step in understanding and ultimately harnessing it for making bio-based toluene. PMID:27506494

  18. In vitro Characterization of Phenylacetate Decarboxylase, a Novel Enzyme Catalyzing Toluene Biosynthesis in an Anaerobic Microbial Community.

    PubMed

    Zargar, K; Saville, R; Phelan, R M; Tringe, S G; Petzold, C J; Keasling, J D; Beller, H R

    2016-08-10

    Anaerobic bacterial biosynthesis of toluene from phenylacetate was reported more than two decades ago, but the biochemistry underlying this novel metabolism has never been elucidated. Here we report results of in vitro characterization studies of a novel phenylacetate decarboxylase from an anaerobic, sewage-derived enrichment culture that quantitatively produces toluene from phenylacetate; complementary metagenomic and metaproteomic analyses are also presented. Among the noteworthy findings is that this enzyme is not the well-characterized clostridial p-hydroxyphenylacetate decarboxylase (CsdBC). However, the toluene synthase under study appears to be able to catalyze both phenylacetate and p-hydroxyphenylacetate decarboxylation. Observations suggesting that phenylacetate and p-hydroxyphenylacetate decarboxylation in complex cell-free extracts were catalyzed by the same enzyme include the following: (i) the specific activity for both substrates was comparable in cell-free extracts, (ii) the two activities displayed identical behavior during chromatographic separation of cell-free extracts, (iii) both activities were irreversibly inactivated upon exposure to O2, and (iv) both activities were similarly inhibited by an amide analog of p-hydroxyphenylacetate. Based upon these and other data, we hypothesize that the toluene synthase reaction involves a glycyl radical decarboxylase. This first-time study of the phenylacetate decarboxylase reaction constitutes an important step in understanding and ultimately harnessing it for making bio-based toluene.

  19. Control by Ethylene of Arginine Decarboxylase Activity in Pea Seedlings and Its Implication for Hormonal Regulation of Plant Growth 1

    PubMed Central

    Apelbaum, Akiva; Goldlust, Arie; Icekson, Isaac

    1985-01-01

    Activity of arginine decarboxylase in etiolated pea seedlings appears 24 hours after seed imbibition, reaches its highest level on the 4th day, and levels off until the 7th day. This activity was found in the apical and subapical tissue of the roots and shoots where intensive DNA synthesis occurs. Exposure of the seedlings to ethylene greatly reduced the specific activity of this enzyme. The inhibition was observed within 30 min of the hormone application, and maximal effect—90% inhibition—after 18 hours. Ethylene at physiological concentrations affected the enzyme activity; 50% inhibitory rate was recorded at 0.12 microliters per liter ethylene and maximal response at 1.2 microliters per liter. Ethylene provoked a 5-fold increase in the Kmapp of arginine decarboxylase for its substrate and reduced the Vmaxapp by 10-fold. However, the enzyme recovered from the inhibition and regained control activity 7 hours after transferral of the seedlings to ethylene-free atmosphere. Reducing the endogenous level of ethylene in the tissue by hypobaric pressure, or by exposure to light, as well as interfering with ethylene action by treatment with silver thiosulfate or 2,5-norbornadiene, caused a gradual increase in the specific activity of arginine decarboxylase in the apical tissue of the etiolated seedlings. On the basis of these findings, the possible control of arginine decarboxylase activity by endogenous ethylene, and its implication for the hormone effect on plant growth, are discussed. PMID:16664464

  20. Temporal relation between the ADC and DC potential responses to transient focal ischemia in the rat: a Markov chain Monte Carlo simulation analysis.

    PubMed

    King, Martin D; Crowder, Martin J; Hand, David J; Harris, Neil G; Williams, Stephen R; Obrenovitch, Tihomir P; Gadian, David G

    2003-06-01

    Markov chain Monte Carlo simulation was used in a reanalysis of the longitudinal data obtained by Harris et al. (J Cereb Blood Flow Metab 20:28-36) in a study of the direct current (DC) potential and apparent diffusion coefficient (ADC) responses to focal ischemia. The main purpose was to provide a formal analysis of the temporal relationship between the ADC and DC responses, to explore the possible involvement of a common latent (driving) process. A Bayesian nonlinear hierarchical random coefficients model was adopted. DC and ADC transition parameter posterior probability distributions were generated using three parallel Markov chains created using the Metropolis algorithm. Particular attention was paid to the within-subject differences between the DC and ADC time course characteristics. The results show that the DC response is biphasic, whereas the ADC exhibits monophasic behavior, and that the two DC components are each distinguishable from the ADC response in their time dependencies. The DC and ADC changes are not, therefore, driven by a common latent process. This work demonstrates a general analytical approach to the multivariate, longitudinal data-processing problem that commonly arises in stroke and other biomedical research.

  1. Identification and measurement of acid (specific) histidine decarboxylase activity in rabbit gastric mucosa: ending an old controversy?

    PubMed

    Neugebauer, E; Lorenz, W

    1985-04-01

    One of the main obstacles in assigning any distinct function to histamine in health and disease was the longlasting controversy on the existence of any physiological, endogenous histamine formation in man and most of the other mammals except the rat. Using a modification of Schayer's isotope dilution method, a renewed attempt was made to identify the very low activities of an acid (specific) histidine decarboxylase in rabbit gastric mucosa capable of producing endogenous histamine in physiological conditions, to develop tests for its identification in crude enzyme extracts and to demonstrate the specificity of the enzymatic assay by excluding any relevant Dopa decarboxylase activity and also nonenzymatic decarboxylation interfering with the determination of acid (specific) histidine decarboxylase. To achieve this aim five tests were developed: In the pH profile (test 1), a pH optimum was found at 7.0 in the presence of a low substrate concentration (1.6 X 10(-6)M L-[ring-2-14C]-histidine). The apparent Michaelis concentration at the pH optimum (test 2) was 1.8 X 10(-4)M, the maximum rate 12.5pmol [14C]histamine formed X min-1. To increase the specificity of inhibition experiments with alpha-methylhistidine and alpha-methyl-L-Dopa a pH profile was determined in the presence of these two enzymatic inhibitors (test 3 and 4). alpha-Methylhistidine was used for a reliable diagnostic confirmation test, alpha-methyl-L-Dopa for a reliable exclusion test. Benzene showed no influence on either blanks or recovery rates, but inhibited the enzymic activity at pH 7.0, not however that of unspecific histidine decarboxylase and hence was very valuable as an additional diagnostic exclusion test (test 5). Although these new tests identifying acid (specific) histidine decarboxylase and demonstrating the specificity of its determination were tedious, despite the use of the modified isotope dilution method, they excluded the presence of any Dopa decarboxylase activity in mixtures with

  2. Expression and stereochemical and isotope effect studies of active 4-oxalocrotonate decarboxylase.

    PubMed

    Stanley, T M; Johnson, W H; Burks, E A; Whitman, C P; Hwang, C C; Cook, P F

    2000-02-01

    4-Oxalocrotonate decarboxylase (4-OD) and vinylpyruvate hydratase (VPH) from Pseudomonas putida mt-2 form a complex that converts 2-oxo-3-hexenedioate to 2-oxo-4-hydroxypentanoate in the catechol meta fission pathway. To facilitate mechanistic and structural studies of the complex, the two enzymes have been coexpressed and the complex has been purified to homogeneity. In addition, Glu-106, a potential catalytic residue in VPH, has been changed to glutamine, and the resulting E106QVPH mutant has been coexpressed with 4-OD and purified to homogeneity. The 4-OD/E106QVPH complex retains full decarboxylase activity, with comparable kinetic parameters to those observed for 4-OD in the wild-type complex, but is devoid of any detectable hydratase activity. Decarboxylation of (5S)-2-oxo-3-[5-D]hexenedioate by either the 4-OD/VPH complex or the mutant complex generates 2-hydroxy-2,4E-[5-D]pentadienoate in D(2)O. Ketonization of 2-hydroxy-2,4-pentadienoate by the wild-type complex is highly stereoselective and results in the formation of 2-oxo-(3S)-[3-D]-4-pentenoate, while the mutant complex generates a racemic mixture. These results indicate that 2-hydroxy-2, 4-pentadienoate is the product of 4-OD and that 2-oxo-4-pentenoate results from a VPH-catalyzed process. On this basis, the previously proposed hypothesis for the conversion of 2-oxo-3-hexenedioate to 2-oxo-4-hydroxypentanoate has been revised [Lian, H., and Whitman, C. P. (1994) J. Am. Chem. Soc. 116, 10403-10411]. Finally, the observed (13)C kinetic isotope effect on the decarboxylation of 2-oxo-3-hexenedioate by the 4-OD/VPH complex suggests that the decarboxylation step is nearly rate-limiting. Because the value is not sensitive to either magnesium or manganese, it is likely that the transition state for carbon-carbon bond cleavage is late and that the metal positions the substrate and polarizes the carbonyl group, analogous to its role in oxalacetate decarboxylase.

  3. Structural determinants for the inhibitory ligands of orotidine-5′-monophosphate decarboxylase

    SciTech Connect

    Meza-Avina, Maria Elena; Wei, Lianhu; Liu, Yan; Poduch, Ewa; Bello, Angelica M.; Mishra, Ram K.; Pai, Emil F.; Kotra, Lakshmi P.

    2010-06-14

    In recent years, orotidine-5{prime}-monophosphate decarboxylase (ODCase) has gained renewed attention as a drug target. As a part of continuing efforts to design novel inhibitors of ODCase, we undertook a comprehensive study of potent, structurally diverse ligands of ODCase and analyzed their structural interactions in the active site of ODCase. These ligands comprise of pyrazole or pyrimidine nucleotides including the mononucleotide derivatives of pyrazofurin, barbiturate ribonucleoside, and 5-cyanouridine, as well as, in a computational approach, 1,4-dihydropyridine-based non-nucleoside inhibitors such as nifedipine and nimodipine. All these ligands bind in the active site of ODCase exhibiting distinct interactions paving the way to design novel inhibitors against this interesting enzyme. We propose an empirical model for the ligand structure for rational modifications in new drug design and potentially new lead structures.

  4. Maternal immune activation alters glutamic acid decarboxylase-67 expression in the brains of adult rat offspring

    PubMed Central

    Cassella, Sarah N.; Hemmerle, Ann M.; Lundgren, Kerstin H.; Kyser, Tara L.; Ahlbrand, Rebecca; Bronson, Stefanie L.; Richtand, Neil M.; Seroogy, Kim B.

    2016-01-01

    Activation of the maternal innate immune system, termed “maternal immune activation” (MIA), represents a common environmental risk factor for schizophrenia. Whereas evidence suggests dysregulation of GABA systems may underlie the pathophysiology of schizophrenia, a role for MIA in alteration of GABAergic systems is less clear. Here, pregnant rats received either the viral mimetic polyriboinosinic-polyribocytidilic acid or vehicle injection on gestational day 14. Glutamic acid decarboxylase-67 (GAD67) mRNA expression was examined in male offspring at postnatal day (P)14, P30 and P60. At P60, GAD67 mRNA was elevated in hippocampus and thalamus and decreased in prefrontal cortex of MIA offspring. MIA-induced alterations in GAD expression could contribute to the pathophysiology of schizophrenia. PMID:26830319

  5. Crystallization and preliminary X-ray analysis of the inducible lysine decarboxylase from Escherichia coli

    SciTech Connect

    Alexopoulos, E.; Kanjee, U.; Snider, J.; Houry, W.A.; Pai, E.F.

    2010-02-11

    The decameric inducible lysine decarboxylase (LdcI) from Escherichia coli has been crystallized in space groups C2 and C222{sub 1}; the Ta{sub 6}Br{sub 12}{sup 2+} cluster was used to derivatize the C2 crystals. The method of single isomorphous replacement with anomalous scattering (SIRAS) as implemented in SHELXD was used to solve the Ta{sub 6}Br{sub 12}{sup 2+}-derivatized structure to 5 {angstrom} resolution. Many of the Ta{sub 6}Br{sub 12}{sup 2+}-binding sites had twofold and fivefold noncrystallographic symmetry. Taking advantage of this feature, phase modification was performed in DM. The electron-density map of LdcI displays many features in agreement with the low-resolution negative-stain electron-density map [Snider et al. (2006), J. Biol. Chem. 281, 1532-1546].

  6. Histidine decarboxylase deficiency causes Tourette syndrome: parallel findings in humans and mice

    PubMed Central

    Baldan, Lissandra Castellan; Rapanelli, Maximiliano; Crowley, Michael; Anderson, George M.; Loring, Erin; Gorczyca, Roxanne; Billingslea, Eileen; Wasylink, Suzanne; Panza, Kaitlyn E.; Ercan-Sencicek, A. Gulhan; Krusong, Kuakarun; Leventhal, Bennett L.; Ohtsu, Hiroshi; Bloch, Michael H.; Hughes, Zoë A.; Krystal, John H.; Mayes, Linda; de Araujo, Ivan; Ding, Yu-Shin; State, Matthew W.; Pittenger, Christopher

    2013-01-01

    Tourette syndrome (TS) is characterized by tics, sensorimotor gating deficiencies, and abnormalities of cortico-basal ganglia circuits. A mutation in histidine decarboxylase (Hdc), the key enzyme for the biosynthesis of histamine (HA), has been implicated as a rare genetic cause. Hdc knockout mice exhibited potentiated tic-like stereotypies, recapitulating core phenomenology of TS; these were mitigated by the dopamine D2 antagonist haloperidol, a proven pharmacotherapy, and by HA infusion into the brain. Prepulse inhibition was impaired in both mice and humans carrying Hdc mutations. HA infusion reduced striatal dopamine (DA) levels; in Hdc knockout mice, striatal DA was increased and the DA-regulated immediate early gene Fos was upregulated. Dopamine D2/D3 receptor binding was altered both in mice and in humans carrying the Hdc mutation. These data confirm HDC deficiency as a rare cause of TS and identify histamine-dopamine interactions in the basal ganglia as an important locus of pathology. PMID:24411733

  7. Extralimbic autoimmune encephalitis associated with glutamic acid decarboxylase antibodies: an underdiagnosed entity?

    PubMed

    Najjar, Souhel; Pearlman, Daniel; Najjar, Amanda; Ghiasian, Vahid; Zagzag, David; Devinsky, Orrin

    2011-07-01

    Nonparaneoplastic glutamic acid decarboxylase antibody (GADAb)-related autoimmune encephalitis is a syndrome characterized by refractory seizures, progressive cognitive deficits, and psychiatric manifestations. The limbic subtype is well described, has characteristic affective and memory disturbances, and typical mesial temporal MRI abnormalities. We found only one single case report of the extralimbic subtype. We report clinical, radiological, and pathological findings of two additional cases with contrast-enhancing lesions. One of our cases presented as vasculitis, and the other imitated a tumor. Pathological evidence of both vasculitis and encephalitis has never been previously reported in any inflammatory condition affecting the brain. Our cases confirm prior reports that immune therapy can better control seizures associated with GADAb autoimmune encephalitis, and support the rationale for assaying for GADAb titers in patients with etiologically unclear extralimbic lesions and refractory epilepsy, independent of seizure types.

  8. Identification and molecular cloning of glutamate decarboxylase gene from Lactobacillus casei

    PubMed Central

    Tavakoli, Yasaman; Esmaeili, Abolghasem; Rabbani, Mohammad

    2015-01-01

    Gamma-amino butyric acid (GABA) possesses several physiological functions such as neurotransmission, induction of hypotension, diuretic and tranquilizer effects. Production of GABA-enriched products by lactic acid bacteria has been a focus of different researches in recent years because of their safety and health-promoting specifities. In this study, glutamate decarboxylase (gad) gene of a local strains Lactobacillus casei was identified and cloned. In order to clone the gad gene from this strain, the PCR was carried out using primers designed based on conserved regions. The PCR product was purified and ligated into PGEM-T vector. Comparison of obtained sequences shows that this fragment codes the pyridoxal 5′-phosphate binding region. This strain could possibly be used for the industrial GABA production and also for development of functional fermented foods. Gad gene manipulation can also either decrease or increase the activity of enzyme in bacteria. PMID:27844008

  9. Molecular mechanism of allosteric substrate activation in a thiamine diphosphate-dependent decarboxylase.

    PubMed

    Versées, Wim; Spaepen, Stijn; Wood, Martin D H; Leeper, Finian J; Vanderleyden, Jos; Steyaert, Jan

    2007-11-30

    Thiamine diphosphate-dependent enzymes are involved in a wide variety of metabolic pathways. The molecular mechanism behind active site communication and substrate activation, observed in some of these enzymes, has since long been an area of debate. Here, we report the crystal structures of a phenylpyruvate decarboxylase in complex with its substrates and a covalent reaction intermediate analogue. These structures reveal the regulatory site and unveil the mechanism of allosteric substrate activation. This signal transduction relies on quaternary structure reorganizations, domain rotations, and a pathway of local conformational changes that are relayed from the regulatory site to the active site. The current findings thus uncover the molecular mechanism by which the binding of a substrate in the regulatory site is linked to the mounting of the catalytic machinery in the active site in this thiamine diphosphate-dependent enzyme.

  10. Some Aspects of Yeast Anaerobic Metabolism Examined by the Inhibition of Pyruvate Decarboxylase

    NASA Astrophysics Data System (ADS)

    Martin, Earl V.

    1998-10-01

    Incubation of yeast cells with various sugars in aqueous alkaline phosphate solutions under anaerobic conditions results in the accumulation of pyruvate in the cell medium after short periods of up to 15 minutes. This accumulation of pyruvate as the end product of glycolysis results from the inhibition of pyruvate decarboxylase under the conditions. This pyruvate production can be readily measured in the cell-free medium by a spectrophotometric assay using lactic dehydrogenase and NADH. The production of pyruvate can be directly related to the ability of the yeast cells to metabolize particular carbon sources provided. Comparison of pyruvate production by yeast from a variety of common sugars, for example, provides students with a means to assess what sugars are readily utilized by this organism. An additional advantage for student laboratory studies is the availability of Sacchromyces cerevisiae at minimal cost as dry granules which are easily weighed and quickly activated.

  11. Improvement of the Process Stability of Arylmalonate Decarboxylase by Immobilization for Biocatalytic Profen Synthesis

    PubMed Central

    Aßmann, Miriam; Mügge, Carolin; Gaßmeyer, Sarah Katharina; Enoki, Junichi; Hilterhaus, Lutz; Kourist, Robert; Liese, Andreas; Kara, Selin

    2017-01-01

    The enzyme arylmalonate decarboxylase (AMDase) enables the selective synthesis of enantiopure (S)-arylpropinates in a simple single-step decarboxylation of dicarboxylic acid precursors. However, the poor enzyme stability with a half-life time of about 1.2 h under process conditions is a serious limitation of the productivity, which results in a need for high catalyst loads. By immobilization on an amino C2 acrylate carrier the operational stability of the (S)-selective AMDase variant G74C/M159L/C188G/V43I/A125P/V156L was increased to a half-life of about 8.6 days, which represents a 158-fold improvement. Further optimization was achieved by simple immobilization of the cell lysate to eliminate the cost- and time intensive enzyme purification step. PMID:28360905

  12. Opsoclonus-myoclonus-ataxia syndrome with autoantibodies to glutamic acid decarboxylase.

    PubMed

    Markakis, Ioannis; Alexiou, Eleni; Xifaras, Michael; Gekas, Georgios; Rombos, Antonios

    2008-06-01

    Opsoclonus-myoclonus-ataxia syndrome (OMS) is a rare neurological disorder of probably autoimmune origin. Most cases are associated with a remote neoplasm or a viral infection; however in some instances no underlying aetiology can be demonstrated. We report the presence of anti-glutamic acid decarboxylase antibodies (anti-GAD Abs) in the serum and CSF of a patient with idiopathic OMS. Treatment with intravenous immunoglobulin led to a remarkable clinical improvement with parallel reduction of anti-GAD titers. Anti-GAD Abs have been associated with several neurological syndromes. They could also be responsible for the clinical triad of OMS, by impairing GABAergic transmission in specific brainstem and cerebellar circuits. We propose that testing for anti-GAD Abs should be performed in OMS, especially when no other aetiological association can be demonstrated.

  13. The importance of SERINE DECARBOXYLASE1 (SDC1) and ethanolamine biosynthesis during embryogenesis of Arabidopsis thaliana.

    PubMed

    Yunus, Ian Sofian; Liu, Yu-Chi; Nakamura, Yuki

    2016-11-01

    In plants, ethanolamine is considered a precursor for the synthesis of choline, which is an essential dietary nutrient for animals. An enzyme serine decarboxylase (SDC) has been identified and characterized in Arabidopsis, which directly converts serine to ethanolamine, a precursor to phosphorylethanolamine and its subsequent metabolites in plants. However, the importance of SDC and ethanolamine production in plant growth and development remains unclear. Here, we show that SDC is required for ethanolamine biosynthesis in vivo and essential in plant embryogenesis in Arabidopsis. The knockout of SDC1 caused an embryonic lethal defect due to the developmental arrest of the embryos at the heart stage. During embryo development, the expression was observed at the later stages, at which developmental defect occurred in the knockout mutant. Overexpression of SDC1 in planta increased levels of ethanolamine, phosphatidylethanolamine, and phosphatidylcholine both in leaves and siliques. These results suggest that SDC1 plays an essential role in ethanolamine biosynthesis during the embryogenesis in Arabidopsis.

  14. Regioselective Enzymatic β-Carboxylation of para-Hydroxy- styrene Derivatives Catalyzed by Phenolic Acid Decarboxylases

    PubMed Central

    Wuensch, Christiane; Pavkov-Keller, Tea; Steinkellner, Georg; Gross, Johannes; Fuchs, Michael; Hromic, Altijana; Lyskowski, Andrzej; Fauland, Kerstin; Gruber, Karl; Glueck, Silvia M; Faber, Kurt

    2015-01-01

    We report on a ‘green’ method for the utilization of carbon dioxide as C1 unit for the regioselective synthesis of (E)-cinnamic acids via regioselective enzymatic carboxylation of para-hydroxystyrenes. Phenolic acid decarboxylases from bacterial sources catalyzed the β-carboxylation of para-hydroxystyrene derivatives with excellent regio- and (E/Z)-stereoselectivity by exclusively acting at the β-carbon atom of the C=C side chain to furnish the corresponding (E)-cinnamic acid derivatives in up to 40% conversion at the expense of bicarbonate as carbon dioxide source. Studies on the substrate scope of this strategy are presented and a catalytic mechanism is proposed based on molecular modelling studies supported by mutagenesis of amino acid residues in the active site. PMID:26190963

  15. Genetic Confirmation of the Role of Sulfopyruvate Decarboxylase in Coenzyme M Biosynthesis in Methanococcus maripaludis

    DOE PAGES

    Sarmiento, Felipe; Ellison, Courtney K.; Whitman, William B.

    2013-01-01

    Coenzyme M is an essential coenzyme for methanogenesis. The proposed biosynthetic pathway consists of five steps, of which the fourth step is catalyzed by sulfopyruvate decarboxylase (ComDE). Disruption of the gene comE by transposon mutagenesis resulted in a partial coenzyme M auxotroph, which grew poorly in the absence of coenzyme M and retained less than 3% of the wild type level of coenzyme M biosynthesis. Upon coenzyme M addition, normal growth of the mutant was restored. Moreover, complementation of the mutation with the wild type comE gene in trans restored full growth in the absence of coenzyme M. Thesemore » results confirm that ComE plays an important role in coenzyme M biosynthesis. The inability to yield a complete CoM auxotroph suggests that either the transposon insertion failed to completely inactivate the gene or M. maripaludis possesses a promiscuous activity that partially complemented the mutation.« less

  16. Hydroxydibenzoylmethane induces apoptosis through repressing ornithine decarboxylase in human promyelocytic leukemia HL-60 cells

    PubMed Central

    Wang, Ming-Fu; Liao, Ya-Fan; Hung, Ying-Cheng; Lin, Chih-Li; Hour, Tzyh-Chyuan; Lue, Ko-Huang; Hung, Hui-Chih

    2011-01-01

    Ornithine decarboxylase (ODC) is the rate-limiting enzyme in polyamine biosynthesis and a target for chemoprevention. Hydroxydibenzoylmethane (HDB), a derivative of dibenzoylmethane of licorice, is a promising chemopreventive agent. In this paper, we investigated whether HDB would inhibit the ODC pathway to enhance apoptosis in human promyelocytic leukemia HL-60 cells. We found ODC enzyme activity was reduced during HDB treatment. Overexpression of ODC in HL-60 parental cells could reduce HDB-induced apoptosis, which leads to loss of mitochondrial membrane potential (Δψm), through lessening intracellular ROS. Furthermore, ODC overexpression protected cytochrome c release and the activation of caspase-3 following HDB treatment. The results demonstrated HDB-induced apoptosis was through a mechanism of down-regulation of ODC and occurred along a ROS-dependent mitochondria-mediated pathway. PMID:21372632

  17. Glutamate alteration of glutamic acid decarboxylase (GAD) in GABAergic neurons: the role of cysteine proteases.

    PubMed

    Monnerie, Hubert; Le Roux, Peter D

    2008-09-01

    Brain cell vulnerability to neurologic insults varies greatly, depending on their neuronal subpopulation. Among cells that survive a pathological insult such as ischemia or brain trauma, some may undergo morphological and/or biochemical changes that could compromise brain function. We previously reported that surviving cortical GABAergic neurons exposed to glutamate in vitro displayed an NMDA receptor (NMDAR)-mediated alteration in the levels of the GABA synthesizing enzyme glutamic acid decarboxylase (GAD65/67) [Monnerie, H., Le Roux, P., 2007. Reduced dendrite growth and altered glutamic acid decarboxylase (GAD) 65- and 67-kDa isoform protein expression from mouse cortical GABAergic neurons following excitotoxic injury in vitro. Exp. Neurol. 205, 367-382]. In this study, we examined the mechanisms by which glutamate excitotoxicity caused a change in cortical GABAergic neurons' GAD protein levels. Removing extracellular calcium prevented the NMDAR-mediated decrease in GAD protein levels, measured using Western blot techniques, whereas inhibiting calcium entry through voltage-gated calcium channels had no effect. Glutamate's effect on GAD protein isoforms was significantly attenuated by preincubation with the cysteine protease inhibitor N-Acetyl-L-Leucyl-L-Leucyl-L-norleucinal (ALLN). Using class-specific protease inhibitors, we observed that ALLN's effect resulted from the blockade of calpain and cathepsin protease activities. Cell-free proteolysis assay confirmed that both proteases were involved in glutamate-induced alteration in GAD protein levels. Together these results suggest that glutamate-induced excitotoxic stimulation of NMDAR in cultured cortical neurons leads to altered GAD protein levels from GABAergic neurons through intracellular calcium increase and protease activation including calpain and cathepsin. Biochemical alterations in surviving cortical GABAergic neurons in various disease states may contribute to the altered balance between excitation

  18. [Cloning, prokaryotic expression and characterization of lysine decarboxylase gene from Huperzia serrata].

    PubMed

    Di, Ci; Li, Jing; Tang, Yuntao; Peng, Qingzhong

    2014-08-01

    Huperzine A is a promising drug to treat Alzheimer's disease (AD). To date, its biosynthetic pathway is still unknown. Lysine decarboxylase (LDC) has been proposed to catalyze the first-step of the biosynthesis of huperzine A. To identify and characterize LDCs from Huperzia serrata, we isolated two LDC fragments (LDC1 and LDC2) from leaves of H. serrata by RT-PCR and then cloned them into pMD 19-T vector. Sequence analysis showed that LDC1 and LDC2 genes shared 95.3% identity and encoded the protein of 212 and 202 amino acid residues respectively. Thus, we ligated LDC genes into pET-32a(+) to obtain recombinant expressing vectors pET-32a(+)/LDC1 and pET-32a(+)/LDC2 respectively. We further introduced two expression vectors into Escherichia coli BL21(DE3) and cultured positive colonies of E. coli in liquid LB medium. After inducing for 4 hours with 260 μg/mL IPTG at 30 degrees C, soluble recombinant Trx-LDC1 and Trx-LDC2 were obtained and isolated for purification using a Ni-NTA affinity chromatography. We incubated purified recombinant proteins with L-lysine in the enzyme reaction buffer at 37 degrees C and then derived the reaction products using dansyl chloride. It was found that both Trx-LDC1 and Trx-LDC2 had decarboxylase activity, could convert L-lysine into cadaverine by way of thin layer chromatography assay. Further, bioinformatics analysis indicated that deduced LDC1 and LDC2 had different physicochemical properties, but similar secondary and three-dimensional structures.

  19. A 128-ch Δ-Σ ADC based mixed signal IC for full digital beamforming Wireless handheld Ultrasound imaging system.

    PubMed

    Chirala, Mohan K; Phuong Huynh; Jaeyoung Ryu; Young-Hwan Kim

    2015-08-01

    This paper reports a massively integrated Δ-Σ ADC based mixed signal chipset for a handheld Wireless Ultrasound imaging system. The IC has been fabricated in a standard 0.13 μm 1.5V 7M2F CMOS process with 128 parallel channels containing Delta-Sigma (Δ-Σ) ADCs, Anti-aliasing filter, Decimation filters, Serializers and LVDS drivers. The entire chip is SPI controlled and allows group-level power control through an FPGA. The IC measures 15 × 15 mm and dissipates around ~ 4.6 W of power, with 12-bit resolution at 20 Msps sample rate. The chip was packaged in a thermally stable BGA package and demonstrated in a handheld ultrasound battery operated system with complete digital beamforming.

  20. Thrombotic Thrombocytopenic Purpura with Reversible Neurological Features: Brain Diffusion MRI with ADC Map, Spect and EEG Findings. A Case Report.

    PubMed

    Yerdelen, D; Göksel, B K; Yıldırım, T; Karataş, M; Karaca, S; Reyhan, M; Ozdoğu, H

    2006-11-30

    Although nervous system involvement is common in thrombotic thrombocytopenic purpura (TTP), abnormalities on computerized tomography, magnetic resonance imaging and electroencephalography are not encountered so frequently and if present, these abnormalities are often reversible. We describe a 39-year-old woman with recurring transient focal neurological findings found to have laboratory findings consistent with TTP. In cerebral diffusion weighted images (DWI), diffuse cortical hyperintensity was noted in right frontal lobe, but the ADC (apparent diffusion coefficient) map was normal. Electroencephalography demonstrated lateralized slowing and repeated DWI showed diffuse cortical hyperintensity in the right hemisphere. SPECT showed luxury perfusion in the right hemisphere areas. The patient's condition resolved with plasmapheresis. Our patient illustrates that diffuse hemispheric involvement can be seen in DWI and EEG, and SPECT may show luxury perfusion after resolution of neurological findings in TTP cases. To our knowledge, this is the first TTP case in which the ADC map was normal.

  1. A Low-Power Reduced Kick-Back Comparator with Improved Calibration for High-Speed Flash ADCs

    NASA Astrophysics Data System (ADS)

    Torfs, Guy; Li, Zhisheng; Bauwelinck, Johan; Yin, Xin; Vandewege, Jan; Plas, Geert Van Der

    A novel low-power kick-back reduced comparator for use in high-speed flash analog-to-digital converters (ADC) is presented. The proposed comparator combines cascode transistors to reduce the kick-back noise with a built-in threshold voltage to remove the static power consumption of a reference. Without degrading other figures, the kick-back noise is reduced by a factor 8, compared to a previous design without cascode transistors. An improved calibration structure is also proposed to improve linearity when used in an ADC. Simulated in a standard CMOS technology the comparator consumes 106.5µW at 1.8V power supply and 1GHz clock frequency.

  2. A 10-b 50-MS/s 820- μW SAR ADC With On-Chip Digital Calibration.

    PubMed

    Yoshioka, Masato; Ishikawa, Kiyoshi; Takayama, Takeshi; Tsukamoto, Sanroku

    2010-12-01

    This 10-b 50-MSamples/s SAR analog-to-digital converter (ADC) features on-chip digital calibration techniques, comparator offset cancellation, a capacitor digital-to-analog converter (CDAC) linearity calibration, and internal clock control to compensate for PVT variations. A split-CDAC reduces the exponential increase in the number of unit capacitors needed and enables the input load capacitance to be as small as the kT/C noise restriction. The prototype fabricated in 65 nm 1P7M complementary metal-oxide semiconductor with MIM capacitor achieves 56.6 dB SNDR at 50-MSamples/s, 25-MHz input frequency and consumes 820 μW from a 1.0-V supply, including the digital calibration circuits. The figure of merit was 29.7 fJ/conversion-step under the Nyquist condition. The ADC occupied an active area of 0.039 mm(2) .

  3. Nonlinear distortions in silicon microring resonator filters and their impact on integrated photonic ADCs

    NASA Astrophysics Data System (ADS)

    Al Qubaisi, Kenaish; Khilo, Anatol

    2016-03-01

    We present a dynamic model based on temporal coupled-mode theory to model microring resonators considering silicon nonlinearities. By taking into account the vectorial nature of the optical modes propagating in strongly confining silicon waveguides, we introduce effective areas for two-photon absorption (TPA) and free-carrier distribution in order to adapt the rate equation describing the generation of free-carriers due to TPA and Sorefs equations for silicon waveguides. The performance of optical systems utilizing microring resonators can be degraded due to its nonlinear response. In this paper, we investigate the impact of silicon nonlinearities in microring resonators on the effective number of bits (ENOB) in integrated photonic analog-to-converters (ADCs). This is done by analyzing the nonlinear response of a first-order microring drop filter to a modulated optical pulse train. The dependence of the nonlinear response of the microring resonator, embodied in the input pulse energy vs output pulse energy, and the maximum ENOB on various filter and input pulse train parameters is analyzed by varying the finesse, microring waveguide geometry, modulation index, and average pulse energy.

  4. LDRD final report: photonic analog-to-digital converter (ADC) technology

    SciTech Connect

    Bowers, M; Deri, B; Haigh, R; Lowry, M; Sargis, P; Stafford, R; Tong, T

    1999-02-18

    We report on an LDRD seed program of novel technology development (started by an FY98 Engineering Tech-base project) that will enable extremely high-fidelity analog-to-digital converters for a variety of national security missions. High speed (l0+ GS/s ), high precision (l0+ bits) ADC technology requires extremely short aperture times ({approx}1ps ) with very low jitter requirements (sub 10fs ). These fundamental requirements, along with other technological barriers, are difficult to realize with electronics: However, we outline here, a way to achieve these timing apertures using a novel multi-wavelength optoelectronic short-pulse optical source. Our approach uses an optoelectronic feedback scheme with high optical Q to produce an optical pulse train with ultra-low jitter ( sub 5fs) and high amplitude stability (<10{sup 10}). This approach requires low power and can be integrated into an optoelectronic integrated circuit to minimize the size. Under this seed program we have demonstrated that the optical feedback mechanism can be used to generate a high Q resonator. This has reduced the technical risk for further development, making it an attractive candidate for outside funding.

  5. Electron momentum spectroscopy of norbornadiene at the benchmark ADC(3) level.

    PubMed

    Morini, Filippo; Hajgató, Balázs; Deleuze, Michael S

    2010-09-02

    An extensive study, throughout the valence region, of the electronic structure, ionization spectrum, and electron momentum distributions of norbornadiene is presented, on the ground of accurate calculations of valence one-electron and shake-up ionization energies and of the related Dyson orbitals, using one-particle Green's function (1p-GF) theory in conjunction with the so-called third-order algebraic diagrammatic construction scheme [ADC(3)]. Comparison is made with results obtained from standard (B3LYP) Kohn-Sham orbitals and measurements employing electron momentum spectroscopy, taking into account the contamination of inner- and outer-valence spectral bands by numerous shake-up states. Four relatively intense shake-up lines at 12.1, 16.4, 17.6, and 17.8 eV are found to yield recognizable spectral fingerprints in the EMS experiments. Valence bands at electron binding energies larger than 20 eV are subject to a complete breakdown of the orbital picture of ionization.

  6. Measurement Of Neutron Radius In Lead By Parity Violating Scattering Flash ADC DAQ

    SciTech Connect

    Ahmed, Zafar

    2012-06-01

    This dissertation reports the experiment PREx, a parity violation experiment which is designed to measure the neutron radius in 208Pb. PREx is performed in hall A of Thomas Jefferson National Accelerator Facility from March 19th to June 21st. Longitudionally polarized electrons at energy 1 GeV scattered at and angle of θlab = 5.8 ° from the Lead target. Beam corrected pairty violaing counting rate asymmetry is (Acorr= 594 ± 50(stat) ± 9(syst))ppb at Q2 = 0.009068GeV 2. This dissertation also presents the details of Flash ADC Data Acquisition(FADC DAQ) system for Moller polarimetry in Hall A of Thomas Jefferson National Accelerator Facility. The Moller polarimeter measures the beam polarization to high precision to meet the specification of the PREx(Lead radius experiment). The FADC DAQ is part of the upgrade of Moller polarimetery to reduce the systematic error for PREx. The hardware setup and the results of the FADC DAQ analysis are presented

  7. The Combined Performance of ADC, CSF CXCL-13, and CSF IL-10 in the Diagnosis of Central Nervous System Lymphoma

    PubMed Central

    Mabray, Marc C.; Barajas, Ramon F.; Villanueva-Meyer, Javier E.; Zhang, Chiyuan A.; Valles, Francisco E.; Rubenstein, James L.; Cha, Soonmee

    2015-01-01

    Background and Purpose CXC chemokine ligand 13 (CXCL-13) and interleukin 10 (IL-10) have emerged as CSF biomarkers for the diagnosis of CNS Lymphoma (CNSL). Our hypothesis is that the combined use of ADC, CXCL-13, and IL-10 will result in increased diagnostic performance as compared to the use of ADC values alone. Materials and Methods 87 patients were included in this HIPAA and IRB compliant study including 43 with CNSL and 44 without CNSL (21 metastases, 14 high-grade gliomas, 9 tumefactive demyelinating lesions) who had undergone CSF proteomic analysis and had a new enhancing mass on brain MRI. Average ADC (ADCavg) was derived by contouring the contrast enhancing tumor volume. Group means were compared via t-tests for ADCavg, CXCL-13, and IL-10. Receiver operating characteristic (ROC) analysis was performed for each individual variable. Multiple variable logistic regression with ROC was performed and the multiple variable ROC was compared to single variable ROCs. Results ADCavg was lower and CSF CXCL-13 and IL-10 values higher in CNSL (p<0.001). Areas under the curve (AUC) ranged from 0.739-0.832 for single variable ROC. Multiple variable logistic regression yielded statistically significant individual effects for all three variables in a combined model. Multiple variable ROC (AUC 0.928) demonstrated statistically significantly superior diagnostic performance as compared to the use of single variables alone. Conclusion The combined use of ADC, CSF CXCL-13, and IL-10 results in increased diagnostic performance for the diagnosis of CNSL. This highlights the importance of CSF analysis when the diagnosis of CNSL is considered based on MRI. PMID:26381553

  8. A low-power 10-bit 250 MS/s dual-channel pipeline ADC in 0.18 μm CMOS

    NASA Astrophysics Data System (ADS)

    Fan, Q.; Chen, J.; Wen, X.; Feng, Y.; Tang, Y.; Zuo, Z.; Gong, D.; Liu, T.; Ye, J.

    2017-02-01

    This paper presents a 10-bit 250-MS/s time-interleaved pipelined analog-to-digital data converter (ADC). A distributed clocking scheme is developed to eliminate timing skew between channels without introducing load capacitance to the driving buffer of the ADC. The channel offset and gain mismatch errors are calibrated in digital domain. In addition, a switch-embedded opamp-sharing technique is developed to reduce the ADC power consumption and eliminate the memory effect. The simulated SNDR and SFDR are 61.84 dB and 78.2 dB, respectively. The ADC core consumes 28 mW under a 1.8 V supply at 250 MS/s sampling rate.

  9. Response monitoring and action limits: use of ADC numbers in understanding the operational characteristics of the Beckman ASTRA chemistry system.

    PubMed

    Kessner, A; Burnett, R W; Bowers, G N

    1981-07-01

    The Beckman ASTRA is a microprocessor-controlled multichannel chemistry analyzer. The output of each module is available to the operator as analog-to-digital conversion (ADC) numbers, which we record during each calibration (Response Monitoring). After so studying three instruments for a total of 33 months, we have established limits for Calibrator ADC numbers that indicate possible operator action (Action Limits). These Action Limits are tighter than the microprocessor's programmed limits, and alert the operator to short- and long-term drift. These tighter limits warn of (a) impending failure of the instrument to calibrate or (b) possible inaccuracies in results for patients. We have instituted changes in preventive maintenance based on our studies of each module's operational characteristics, and have replaced electrodes that failed to meet our Response Monitoring specifications. Response Monitoring and Action Limits based upon ADC numbers have significantly enhanced our understanding of the ASTRA system and thus improved its operational efficiency and analytical reliability. Estimates of precision and accuracy (true value) were satisfactory in comparison to our prior single-channel continuous-flow and flame photometry analytical measurement systems.

  10. Power and area efficient 4-bit column-level ADC in a CMOS pixel sensor for the ILD vertex detector

    NASA Astrophysics Data System (ADS)

    Zhang, L.; Morel, F.; Hu-Guo, Ch; Hu, Y.

    2013-01-01

    A 48 × 64 pixels prototype CMOS pixel sensor (CPS) integrated with 4-bit column-level, self triggered ADCs for the outer layers of the ILD vertex detector (VTX) was developed and fabricated in a 0.35 μm CMOS process with a pixel pitch of 35 μm. The pixel concept combines in-pixel amplification with a correlated double sampling (CDS) operation. The ADCs accommodating the pixel read out in a rolling shutter mode complete the conversion by performing a multi-bit/step approximation. The design was optimised for power saving at sampling frequency. The prototype sensor is currently at the stage of being started testing and evaluation. So what is described is based on post simulation results rather than test data. This 4-bit ADC dissipates, at a 3-V supply and 6.25-MS/s sampling rate, 486 μW in its inactive mode, which is by far the most frequent. This value rises to 714 μW in case of the active mode. Its footprint amounts to 35 × 545 μm2.

  11. SEMICONDUCTOR INTEGRATED CIRCUITS: A low power cyclic ADC design for a wireless monitoring system for orthopedic implants

    NASA Astrophysics Data System (ADS)

    Yi, Chen; Fule, Li; Hong, Chen; Chun, Zhang; Zhihua, Wang

    2009-08-01

    This paper presents a low power cyclic analog-to-digital convertor (ADC) design for a wireless monitoring system for orthopedic implants. A two-stage cyclic structure including a single to differential converter, two multiplying DAC functional blocks (MDACs) and some comparators is adopted, which brings moderate speed and moderate resolution with low power consumption. The MDAC is implemented with the common switched capacitor method. The 1.5-bit stage greatly simplifies the design of the comparator. The operational amplifier is carefully optimized both in schematic and layout for low power and offset. The prototype chip has been fabricated in a United Microelectronics Corporation (UMC) 0.18-μm 1P6M CMOS process. The core of the ADC occupies only 0.12 mm2. With a 304.7-Hz input and 4-kHz sampling rate, the measured peak SNDR and SFDR are 47.1 dB and 57.8 dBc respectively and its DNL and INL are 0.27 LSB and 0.3 LSB, respectively. The power consumption of the ADC is only 12.5 μW in normal working mode and less than 150 nW in sleep mode.

  12. Adjustable Nyquist-rate System for Single-Bit Sigma-Delta ADC with Alternative FIR Architecture

    NASA Astrophysics Data System (ADS)

    Frick, Vincent; Dadouche, Foudil; Berviller, Hervé

    2016-09-01

    This paper presents a new smart and compact system dedicated to control the output sampling frequency of an analogue-to-digital converters (ADC) based on single-bit sigma-delta (ΣΔ) modulator. This system dramatically improves the spectral analysis capabilities of power network analysers (power meters) by adjusting the ADC's sampling frequency to the input signal's fundamental frequency with a few parts per million accuracy. The trade-off between straightforwardness and performance that motivated the choice of the ADC's architecture are preliminary discussed. It particularly comes along with design considerations of an ultra-steep direct-form FIR that is optimised in terms of size and operating speed. Thanks to compact standard VHDL language description, the architecture of the proposed system is particularly suitable for application-specific integrated circuit (ASIC) implementation-oriented low-power and low-cost power meter applications. Field programmable gate array (FPGA) prototyping and experimental results validate the adjustable sampling frequency concept. They also show that the system can perform better in terms of implementation and power capabilities compared to dedicated IP resources.

  13. A CMOS switch-capacitor 14-bit 100 Msps pipeline ADC with over 90 dB SFDR

    NASA Astrophysics Data System (ADS)

    Cai, Hua; Li, Ping

    2013-01-01

    This article presents a design of 14-bit 100 Msamples/s pipelined analog-to-digital converter (ADC) implemented in 0.18 µm CMOS. A charge-sharing correction (CSC) is proposed to remove the input-dependent charge-injection, along with a floating-well bulk-driven technique, a fast-settling reference generator and a low-jitter clock circuit, guaranteeing the high dynamic performance of the ADC. A scheme of background calibration minimises the error due to the capacitor mismatch and opamp non-ideality, ensuring the overall linearity. The measured results show that the prototype ADC achieves spurious-free dynamic range (SFDR) of 91 dB, signal-to-noise-and-distortion ratio (SNDR) of 73.1 dB, differential nonlinearity (DNL) of +0.61/-0.57 LSB and integrated nonlinearity (INL) of +1.1/-1.0 LSB at 30 MHz input and maintains over 78 dB SFDR and 65 dB SNDR up to 425 MHz, consuming 223 mW totally.

  14. Design and simulation of a 12-bit, 40 MSPS asynchronous SAR ADC for the readout of PMT signals

    NASA Astrophysics Data System (ADS)

    Liu, Jian-Feng; Zhao, Lei; Qin, Jia-Jun; Yang, Yun-Fan; Yu, Li; Liang, Yu; Liu, Shu-Bin; An, Qi

    2016-11-01

    High precision and large dynamic range measurement are required in the readout systems for the Water Cherenkov Detector Array (WCDA) in the Large High Altitude Air Shower Observatory (LHAASO). This paper presents a prototype of a 12-bit 40 MSPS Analog-to-Digital Converter (ADC) Application Specific Integrated Circuit (ASIC) designed for the readout of the LHAASO WCDA. Combining this ADC and the front-end ASIC finished in our previous work, high precision charge measurement can be achieved based on the digital peak detection method. This ADC is implemented based on a power-efficient Successive Approximation Register (SAR) architecture, which incorporates key parts such as a Capacitive Digital-to-Analog Converter (CDAC), dynamic comparator and asynchronous SAR control logic. The simulation results indicate that the Effective Number Of Bits (ENOB) with a sampling rate of 40 MSPS is better than 10 bits in an input frequency range below 20 MHz, while its core power consumption is 6.6 mW per channel. The above results are good enough for the readout requirements of the WCDA. Supported by Knowledge Innovation Program of the Chinese Academy of Sciences (KJCX2-YW-N27), CAS Center for Excellence in Particle Physics (CCEPP)

  15. A 12-bit, 1 MS/s SAR-ADC for a CZT-based multi-channel gamma-ray imager using a new digital calibration method

    NASA Astrophysics Data System (ADS)

    Liu, W.; Wei, T.; Yang, L.; Hu, Y.

    2016-03-01

    The successive approximation register-analog to digital converter (SAR-ADC) is widely used in the CdZnTe-based gamma-ray imager because of its outstanding characteristics of low power consumption, relatively high resolution, and small die size. This study proposes a digital bit-by-bit calibration method using an input ramp signal to further improve the conversion precision and power consumption of an SAR-ADC. The proposed method is based on the sub-radix-2 redundant architecture and the perturbation technique. The proposed calibration algorithm is simpler, more stable, and faster than traditional approaches. The prototype chip of the 12-bit, 1 MS/s radiation-hardened SAR-ADC has been designed and fabricated using the TSMC 0.35 μm 2P4M CMOS process. This SAR-ADC consumes 3 mW power and occupies a core area of 856× 802μm2. The digital bit-by-bit calibration algorithm is implemented via MATLAB for testing flexibility. The effective number of bits for this digitally calibrated SAR-ADC reaches 11.77 bits. The converter exhibits high conversion precision, low power consumption, and radiation-hardened design. Therefore, this SAR-ADC is suitable for multi-channel gamma-ray imager applications.

  16. A 12-bit high-speed column-parallel two-step single-slope analog-to-digital converter (ADC) for CMOS image sensors.

    PubMed

    Lyu, Tao; Yao, Suying; Nie, Kaiming; Xu, Jiangtao

    2014-11-17

    A 12-bit high-speed column-parallel two-step single-slope (SS) analog-to-digital converter (ADC) for CMOS image sensors is proposed. The proposed ADC employs a single ramp voltage and multiple reference voltages, and the conversion is divided into coarse phase and fine phase to improve the conversion rate. An error calibration scheme is proposed to correct errors caused by offsets among the reference voltages. The digital-to-analog converter (DAC) used for the ramp generator is based on the split-capacitor array with an attenuation capacitor. Analysis of the DAC's linearity performance versus capacitor mismatch and parasitic capacitance is presented. A prototype 1024 × 32 Time Delay Integration (TDI) CMOS image sensor with the proposed ADC architecture has been fabricated in a standard 0.18 μm CMOS process. The proposed ADC has average power consumption of 128 μW and a conventional rate 6 times higher than the conventional SS ADC. A high-quality image, captured at the line rate of 15.5 k lines/s, shows that the proposed ADC is suitable for high-speed CMOS image sensors.

  17. Evaluation of the Substrate Scope of Benzoic Acid (De)carboxylases According to Chemical and Biochemical Parameters.

    PubMed

    Pesci, Lorenzo; Kara, Selin; Liese, Andreas

    2016-10-04

    The enzymatic carboxylation of phenolic compounds has been attracting increasing interest in recent years, owing to its regioselectivity and technical potential as a biocatalytic equivalent for the Kolbe-Schmitt reaction. Mechanistically the reaction was demonstrated to occur through electrophilic aromatic substitution/water elimination with bicarbonate as a cosubstrate. The effects of the substituents on the phenolic ring have not yet been elucidated in detail, but this would give detailed insight into the substrate-activity relationship and would provide predictability for the acceptance of future substrates. In this report we show how the kinetic and (apparent) thermodynamic behavior can be explained through the evaluation of linear free energy relationships based on electronic, steric, and geometric parameters and through the consideration of enzyme-ligand interactions. Moreover, the similarity between the benzoic acid decarboxylases and the amidohydrolases superfamily is investigated, and promiscuous hydrolytic activity of the decarboxylase in the context of the hydrolysis of an activated ester bond has been established.

  18. Threonine 57 is required for the post-translational activation of Escherichia coli aspartate α-decarboxylase

    PubMed Central

    Webb, Michael E.; Yorke, Briony A.; Kershaw, Tom; Lovelock, Sarah; Lobley, Carina M. C.; Kilkenny, Mairi L.; Smith, Alison G.; Blundell, Tom L.; Pearson, Arwen R.; Abell, Chris

    2014-01-01

    Aspartate α-decarboxylase is a pyruvoyl-dependent decarboxylase required for the production of β-alanine in the bacterial pantothenate (vitamin B5) biosynthesis pathway. The pyruvoyl group is formed via the intramolecular rearrangement of a serine residue to generate a backbone ester intermediate which is cleaved to generate an N-terminal pyruvoyl group. Site-directed mutagenesis of residues adjacent to the active site, including Tyr22, Thr57 and Tyr58, reveals that only mutation of Thr57 leads to changes in the degree of post-translational activation. The crystal structure of the site-directed mutant T57V is consistent with a non-rearranged backbone, supporting the hypothesis that Thr57 is required for the formation of the ester intermediate in activation. PMID:24699660

  19. Threonine 57 is required for the post-translational activation of Escherichia coli aspartate α-decarboxylase.

    PubMed

    Webb, Michael E; Yorke, Briony A; Kershaw, Tom; Lovelock, Sarah; Lobley, Carina M C; Kilkenny, Mairi L; Smith, Alison G; Blundell, Tom L; Pearson, Arwen R; Abell, Chris

    2014-04-01

    Aspartate α-decarboxylase is a pyruvoyl-dependent decarboxylase required for the production of β-alanine in the bacterial pantothenate (vitamin B5) biosynthesis pathway. The pyruvoyl group is formed via the intramolecular rearrangement of a serine residue to generate a backbone ester intermediate which is cleaved to generate an N-terminal pyruvoyl group. Site-directed mutagenesis of residues adjacent to the active site, including Tyr22, Thr57 and Tyr58, reveals that only mutation of Thr57 leads to changes in the degree of post-translational activation. The crystal structure of the site-directed mutant T57V is consistent with a non-rearranged backbone, supporting the hypothesis that Thr57 is required for the formation of the ester intermediate in activation.

  20. Computational, structural, and kinetic evidence that Vibrio vulnificus FrsA is not a cofactor-independent pyruvate decarboxylase.

    PubMed

    Kellett, Whitney F; Brunk, Elizabeth; Desai, Bijoy J; Fedorov, Alexander A; Almo, Steven C; Gerlt, John A; Rothlisberger, Ursula; Richards, Nigel G J

    2013-03-19

    The fermentation-respiration switch (FrsA) protein in Vibrio vulnificus was recently reported to catalyze the cofactor-independent decarboxylation of pyruvate. We now report quantum mechanical/molecular mechenical calculations that examine the energetics of C-C bond cleavage for a pyruvate molecule bound within the putative active site of FrsA. These calculations suggest that the barrier to C-C bond cleavage in the bound substrate is 28 kcal/mol, which is similar to that estimated for the uncatalyzed decarboxylation of pyruvate in water at 25 °C. In agreement with the theoretical predictions, no pyruvate decarboxylase activity was detected for recombinant FrsA protein that could be crystallized and structurally characterized. These results suggest that the functional annotation of FrsA as a cofactor-independent pyruvate decarboxylase is incorrect.

  1. Computational, Structural and Kinetic Evidence that Vibrio vulnificus FrsA is not a Cofactor-Independent Pyruvate Decarboxylase

    PubMed Central

    Kellett, Whitney F.; Brunk, Elizabeth; Desai, Bijoy J.; Fedorov, Alexander A.; Almo, Steven C.; Gerlt, John A.; Rothlisberger, Ursula; Richards, Nigel G. J.

    2013-01-01

    The fermentation-respiration switch (FrsA) protein in Vibrio vulnificus was recently reported to catalyze the cofactor-independent decarboxylation of pyruvate. We now report QM/MM calculations that examine the energetics of C-C bond cleavage for a pyruvate molecule bound within the putative active site of FrsA. These calculations suggest that the barrier to C-C bond cleavage in the bound substrate is 28 kcal/mol, which is similar to that estimated for the uncatalyzed decarboxylation of pyruvate in water at 25 °C. In agreement with the theoretical predictions, no pyruvate decarboxylase activity was detected for recombinant FrsA protein that could be crystallized and structurally characterized. These results suggest that the functional annotation of FrsA as a cofactor-independent pyruvate decarboxylase is incorrect. PMID:23452154

  2. Effects of down-regulating ornithine decarboxylase upon putrescine-associated metabolism and growth in Nicotiana tabacum L.

    PubMed Central

    Dalton, Heidi L.; Blomstedt, Cecilia K.; Neale, Alan D.; Gleadow, Ros; DeBoer, Kathleen D.; Hamill, John D.

    2016-01-01

    Transgenic plants of Nicotiana tabacum L. homozygous for an RNAi construct designed to silence ornithine decarboxylase (ODC) had significantly lower concentrations of nicotine and nornicotine, but significantly higher concentrations of anatabine, compared with vector-only controls. Silencing of ODC also led to significantly reduced concentrations of polyamines (putrescine, spermidine and spermine), tyramine and phenolamides (caffeoylputrescine and dicaffeoylspermidine) with concomitant increases in concentrations of amino acids ornithine, arginine, aspartate, glutamate and glutamine. Root transcript levels of S-adenosyl methionine decarboxylase, S-adenosyl methionine synthase and spermidine synthase (polyamine synthesis enzymes) were reduced compared with vector controls, whilst transcript levels of arginine decarboxylase (putrescine synthesis), putrescine methyltransferase (nicotine production) and multi-drug and toxic compound extrusion (alkaloid transport) proteins were elevated. In contrast, expression of two other key proteins required for alkaloid synthesis, quinolinic acid phosphoribosyltransferase (nicotinic acid production) and a PIP-family oxidoreductase (nicotinic acid condensation reactions), were diminished in roots of odc-RNAi plants relative to vector-only controls. Transcriptional and biochemical differences associated with polyamine and alkaloid metabolism were exacerbated in odc-RNAi plants in response to different forms of shoot damage. In general, apex removal had a greater effect than leaf wounding alone, with a combination of these injury treatments producing synergistic responses in some cases. Reduced expression of ODC appeared to have negative effects upon plant growth and vigour with some leaves of odc-RNAi lines being brittle and bleached compared with vector-only controls. Together, results of this study demonstrate that ornithine decarboxylase has important roles in facilitating both primary and secondary metabolism in Nicotiana. PMID

  3. Effects of down-regulating ornithine decarboxylase upon putrescine-associated metabolism and growth in Nicotiana tabacum L.

    PubMed

    Dalton, Heidi L; Blomstedt, Cecilia K; Neale, Alan D; Gleadow, Ros; DeBoer, Kathleen D; Hamill, John D

    2016-05-01

    Transgenic plants of Nicotiana tabacum L. homozygous for an RNAi construct designed to silence ornithine decarboxylase (ODC) had significantly lower concentrations of nicotine and nornicotine, but significantly higher concentrations of anatabine, compared with vector-only controls. Silencing of ODC also led to significantly reduced concentrations of polyamines (putrescine, spermidine and spermine), tyramine and phenolamides (caffeoylputrescine and dicaffeoylspermidine) with concomitant increases in concentrations of amino acids ornithine, arginine, aspartate, glutamate and glutamine. Root transcript levels of S-adenosyl methionine decarboxylase, S-adenosyl methionine synthase and spermidine synthase (polyamine synthesis enzymes) were reduced compared with vector controls, whilst transcript levels of arginine decarboxylase (putrescine synthesis), putrescine methyltransferase (nicotine production) and multi-drug and toxic compound extrusion (alkaloid transport) proteins were elevated. In contrast, expression of two other key proteins required for alkaloid synthesis, quinolinic acid phosphoribosyltransferase (nicotinic acid production) and a PIP-family oxidoreductase (nicotinic acid condensation reactions), were diminished in roots of odc-RNAi plants relative to vector-only controls. Transcriptional and biochemical differences associated with polyamine and alkaloid metabolism were exacerbated in odc-RNAi plants in response to different forms of shoot damage. In general, apex removal had a greater effect than leaf wounding alone, with a combination of these injury treatments producing synergistic responses in some cases. Reduced expression of ODC appeared to have negative effects upon plant growth and vigour with some leaves of odc-RNAi lines being brittle and bleached compared with vector-only controls. Together, results of this study demonstrate that ornithine decarboxylase has important roles in facilitating both primary and secondary metabolism in Nicotiana.

  4. Cloning and characterization of a locus encoding an indolepyruvate decarboxylase involved in indole-3-acetic acid synthesis in Erwinia herbicola.

    PubMed Central

    Brandl, M T; Lindow, S E

    1996-01-01

    Erwinia herbicola 299R synthesizes indole-3-acetic acid (IAA) primarily by the indole-3-pyruvic acid pathway. A gene involved in the biosynthesis of IAA was cloned from strain 299R. This gene (ipdC) conferred the synthesis of indole-3-acetaldehyde and tryptophol upon Escherichia coli DH5 alpha in cultures supplemented with L-tryptophan. The deduced amino acid sequence of the gene product has high similarity to that of the indolepyruvate decarboxylase of Enterobacter cloacae. Regions within pyruvate decarboxylases of various fungal and plant species also exhibited considerable homology to portions of this gene. This gene therefore presumably encodes an indolepyruvate decarboxylase (IpdC) which catalyzes the conversion of indole-3-pyruvic acid to indole-3-acetaldehyde. Insertions of Tn3-spice within ipdC abolished the ability of strain 299R to synthesize indole-3-acetaldehyde and tryptophol and reduced its IAA production in tryptophan-supplemented minimal medium by approximately 10-fold, thus providing genetic evidence for the role of the indolepyruvate pathway in IAA synthesis in this strain. An ipdC probe hybridized strongly with the genomic DNA of all E. herbicola strains tested in Southern hybridization studies, suggesting that the indolepyruvate pathway is common in this species. Maximum parsimony analysis revealed that the ipdC gene is highly conserved within this group and that strains of diverse geographic origin were very similar with respect to ipdC. PMID:8900003

  5. Lysine Decarboxylase Catalyzes the First Step of Quinolizidine Alkaloid Biosynthesis and Coevolved with Alkaloid Production in Leguminosae[W][OA

    PubMed Central

    Bunsupa, Somnuk; Katayama, Kae; Ikeura, Emi; Oikawa, Akira; Toyooka, Kiminori; Saito, Kazuki; Yamazaki, Mami

    2012-01-01

    Lysine decarboxylase (LDC) catalyzes the first-step in the biosynthetic pathway of quinolizidine alkaloids (QAs), which form a distinct, large family of plant alkaloids. A cDNA of lysine/ornithine decarboxylase (L/ODC) was isolated by differential transcript screening in QA-producing and nonproducing cultivars of Lupinus angustifolius. We also obtained L/ODC cDNAs from four other QA-producing plants, Sophora flavescens, Echinosophora koreensis, Thermopsis chinensis, and Baptisia australis. These L/ODCs form a phylogenetically distinct subclade in the family of plant ornithine decarboxylases. Recombinant L/ODCs from QA-producing plants preferentially or equally catalyzed the decarboxylation of l-lysine and l-ornithine. L. angustifolius L/ODC (La-L/ODC) was found to be localized in chloroplasts, as suggested by the transient expression of a fusion protein of La-L/ODC fused to the N terminus of green fluorescent protein in Arabidopsis thaliana. Transgenic tobacco (Nicotiana tabacum) suspension cells and hairy roots produced enhanced levels of cadaverine-derived alkaloids, and transgenic Arabidopsis plants expressing (La-L/ODC) produced enhanced levels of cadaverine, indicating the involvement of this enzyme in lysine decarboxylation to form cadaverine. Site-directed mutagenesis and protein modeling studies revealed a structural basis for preferential LDC activity, suggesting an evolutionary implication of L/ODC in the QA-producing plants. PMID:22415272

  6. Lysine decarboxylase catalyzes the first step of quinolizidine alkaloid biosynthesis and coevolved with alkaloid production in leguminosae.

    PubMed

    Bunsupa, Somnuk; Katayama, Kae; Ikeura, Emi; Oikawa, Akira; Toyooka, Kiminori; Saito, Kazuki; Yamazaki, Mami

    2012-03-01

    Lysine decarboxylase (LDC) catalyzes the first-step in the biosynthetic pathway of quinolizidine alkaloids (QAs), which form a distinct, large family of plant alkaloids. A cDNA of lysine/ornithine decarboxylase (L/ODC) was isolated by differential transcript screening in QA-producing and nonproducing cultivars of Lupinus angustifolius. We also obtained L/ODC cDNAs from four other QA-producing plants, Sophora flavescens, Echinosophora koreensis, Thermopsis chinensis, and Baptisia australis. These L/ODCs form a phylogenetically distinct subclade in the family of plant ornithine decarboxylases. Recombinant L/ODCs from QA-producing plants preferentially or equally catalyzed the decarboxylation of L-lysine and L-ornithine. L. angustifolius L/ODC (La-L/ODC) was found to be localized in chloroplasts, as suggested by the transient expression of a fusion protein of La-L/ODC fused to the N terminus of green fluorescent protein in Arabidopsis thaliana. Transgenic tobacco (Nicotiana tabacum) suspension cells and hairy roots produced enhanced levels of cadaverine-derived alkaloids, and transgenic Arabidopsis plants expressing (La-L/ODC) produced enhanced levels of cadaverine, indicating the involvement of this enzyme in lysine decarboxylation to form cadaverine. Site-directed mutagenesis and protein modeling studies revealed a structural basis for preferential LDC activity, suggesting an evolutionary implication of L/ODC in the QA-producing plants.

  7. Brain Extraction in Pediatric ADC Maps, toward Characterizing Neuro-Development in Multi-Platform and Multi-Institution Clinical Images

    PubMed Central

    Ou, Yangming; Gollub, Randy L.; Retzepi, Kallirroi; Reynolds, Nathaniel; Pienaar, Rudolph; Pieper, Steve; Murphy, Shawn N.; Grant, P. Ellen; Zöllei, Lilla

    2015-01-01

    Apparent Diffusion Coefficient (ADC) maps can be used to characterize myelination and to detect abnormalities in the developing brain. However, given the normal variation in regional ADC with myelination, detection of abnormalities is difficult when based singularly on visual assessment. Quantitative and automated analysis of pediatric ADC maps is thus desired but requires accurate brain extraction as the first step. Currently, most existing brain extraction methods are optimized for structural T1-weighted MR images of fully myelinated brains. Due to differences in age and image contrast, these approaches do not translate well to pediatric ADC maps. To address this problem, we present a multi-atlas brain extraction framework that has 1) specificity: designed and optimized specifically for pediatric ADC maps; 2) generality: applicable to multi-platform and multi-institution data, and to subjects at various neuro-developmental stages across the first 6 years of life; 3) accuracy: highly accurate compared to expert annotations; and 4) consistency: consistently accurate regardless of sources of data and ages of subjects. We show how we achieve these goals, via optimizing major components in a multi-atlas brain extraction framework, and via developing and evaluating new criteria for its atlas ranking component. Moreover, we demonstrate that these goals can be achieved with a fixed set of atlases and a fixed set of parameters, which opens doors for our optimized framework to be used in large-scale and multi-institution neuro-developmental and clinical studies. In a pilot study, we use this framework in a dataset containing scanner-generated ADC maps from 308 pediatric patients collected during the course of routine clinical care. Our framework leads to successful quantifications of the changes in whole-brain volumes and mean ADC values across the first 6 years of life. PMID:26260429

  8. Herbacetin is a novel allosteric inhibitor of ornithine decarboxylase with antitumor activity

    PubMed Central

    Lee, Mee-Hyun; Oi, Naomi; Lim, Do Young; Kim, Myoung Ok; Cho, Young-Yeon; Pugliese, Angelo; Shim, Jung-Hyun; Chen, Hanyong; Cho, Eun Jin; Kim, Jong-Eun; Kang, Sun Chul; Paul, Souren; Kang, Hee Eun; Jung, Ji Won; Lee, Sung-Young; Kim, Sung-Hyun; Reddy, Kanamata; Yeom, Young Il; Bode, Ann M; Dong, Zigang

    2015-01-01

    Ornithine decarboxylase (ODC) is a rate-limiting enzyme in the first step of polyamine biosynthesis that is associated with cell growth and tumor formation. Existing catalytic inhibitors of ODC have lacked efficacy in clinical testing or displayed unacceptable toxicity. In this study, we report the identification of an effective and nontoxic allosteric inhibitor of ODC. Using computer docking simulation and an in vitro ODC enzyme assay, we identified herbacetin, a natural compound found in flax and other plants, as a novel ODC inhibitor. Mechanistic investigations defined aspartate 44 in ODC as critical for binding. Herbacetin exhibited potent anticancer activity in colon cancer cell lines expressing high levels of ODC. Intraperitoneal or oral administration of herbacetin effectively suppressed HCT116 xenograft tumor growth and also reduced the number and size of polyps in a mouse model of APC-driven colon cancer (ApcMin/+). Unlike the well established ODC inhibitor DFMO, herbacetin treatment was not associated with hearing loss. Taken together, our findings defined the natural product herbacetin as an allosteric inhibitor of ODC with chemopreventive and antitumor activity in preclinical models of colon cancer, prompting its further investigation in clinical trials. PMID:26676750

  9. Structural requirements for novel coenzyme-substrate derivatives to inhibit intracellular ornithine decarboxylase and cell proliferation.

    PubMed

    Wu, Fang; Gehring, Heinz

    2009-02-01

    Creating transition-state mimics has proven to be a powerful strategy in developing inhibitors to treat malignant diseases in several cases. In the present study, structurally diverse coenzyme-substrate derivatives mimicking this type for pyridoxal 5'-phosphate-dependent human ornithine decarboxylase (hODC), a potential anticancer target, were designed, synthesized, and tested to elucidate the structural requirements for optimal inhibition of intracellular ODC as well as of tumor cell proliferation. Of 23 conjugates, phosphopyridoxyl- and pyridoxyl-L-tryptophan methyl ester (pPTME, PTME) proved significantly more potent in suppression proliferation (IC(50) up to 25 microM) of glioma cells (LN229) than alpha-DL-difluoromethylornithine (DFMO), a medically used irreversible inhibitor of ODC. In agreement with molecular modeling predictions, the inhibitory action of pPTME and PTME toward intracellular ODC of LN229 cells exceeded that of the previous designed lead compound POB. The inhibitory active compounds feature hydrophobic side chain fragments and a kind of polyamine motif (-NH-(CH(X))(4)-NH-). In addition, they induce, as polyamine analogs often do, the activity of the polyamine catabolic enzymes polyamine oxidase and spermine/spermidine N(1)-acetyltransferase up to 250 and 780%, respectively. The dual-action mode of these compounds in LN229 cells affects the intracellular polyamine metabolism and might underlie the more favorable cell proliferation inhibition in comparison with DFMO.

  10. Biochemical evaluation of a parsley tyrosine decarboxylase results in a novel 4-hydroxyphenylacetaldehyde synthase enzyme.

    PubMed

    Torrens-Spence, Michael P; Gillaspy, Glenda; Zhao, Bingyu; Harich, Kim; White, Robert H; Li, Jianyong

    2012-02-10

    Plant aromatic amino acid decarboxylases (AAADs) are effectively indistinguishable from plant aromatic acetaldehyde syntheses (AASs) through primary sequence comparison. Spectroscopic analyses of several characterized AASs and AAADs were performed to look for absorbance spectral identifiers. Although this limited survey proved inconclusive, the resulting work enabled the reevaluation of several characterized plant AAS and AAAD enzymes. Upon completion, a previously reported parsley AAAD protein was demonstrated to have AAS activity. Substrate specificity tests demonstrate that this novel AAS enzyme has a unique substrate specificity towards tyrosine (km 0.46mM) and dopa (km 1.40mM). Metabolite analysis established the abundance of tyrosine and absence of dopa in parsley extracts. Such analysis indicates that tyrosine is likely to be the sole physiological substrate. The resulting information suggests that this gene is responsible for the in vivo production of 4-hydroxyphenylacetaldehyde (4-HPAA). This is the first reported case of an AAS enzyme utilizing tyrosine as a primary substrate and the first report of a single enzyme capable of producing 4-HPAA from tyrosine.

  11. Enhanced histamine production through the induction of histidine decarboxylase expression by phorbol ester in Jurkat cells.

    PubMed

    Nagashima, Yusuke; Kako, Koichiro; Kim, Jun-Dal; Fukamizu, Akiyoshi

    2012-11-01

    Histamine (HA), a mediator of inflammation, type I allergic responses and neurotransmission, is synthesized from L-histidine, the reaction of which is catalyzed by histidine decarboxylase (HDC). HDC has been reported to be induced by various stimuli, not only in mast cells and basophils, but also in T lymphocytes and macrophages. Although its mRNA has been shown to be increased in Jurkat cells when treated with phorbol 12-myristate 13-acetate (TPA), little is known concerning the induced production of HA by HDC. The present study quantified the trace amounts of intracellular HA using ultra-high liquid chromatography in combination with the 6-aminoquinoline carbamate-derivatization technique. To test whether the cellular level of HA is elevated by the induction of HDC in Jurkat cells treated with TPA, the peak corresponding to authentic HA in the cell lysate was fractioned and its molecular weight determined by matrix-assisted laser desorption/ionization quadrupole ion trap time-of-flight mass spectrometry. The results of this study show that the HA level is increased by the induction of HDC expression by TPA in Jurkat cells. Therefore, this method is useful in elucidating the physiological significance of HA production.

  12. Multiple promoter elements govern expression of the human ornithine decarboxylase gene in colon carcinoma cells.

    PubMed Central

    Moshier, J A; Osborne, D L; Skunca, M; Dosescu, J; Gilbert, J D; Fitzgerald, M C; Polidori, G; Wagner, R L; Friezner Degen, S J; Luk, G D

    1992-01-01

    Overexpression of the ornithine decarboxylase (ODC) gene may be important to the development and maintenance of colonic neoplasms, as well as tumors in general. In this study, we examined the promoter elements governing constitutive expression of the human ODC gene in HCT 116 human colon carcinoma cells and, for comparison, K562 human erythro-leukemia cells. It was determined by functional analysis that the promoter elements responsible reside within the 378 bp immediately upstream from the transcription start site. Within this sequence, there are at least three regions that modulate the efficiency of the ODC promoter cooperatively. Both DNA bandshift and footprint assays demonstrated all three regions to be rich in sites that bind to nuclear proteins isolated from HCT 116 and K562 cells; the protein binding pattern of non-transformed, diploid fibroblasts was found to be much less complex. Several of the protein binding sequences have little or no homology to common regulatory elements. We suggest that the constitutive activity of the ODC gene in HCT 116 colon carcinoma cells, and perhaps transformed cells in general, involves a complex interaction of multiple regulatory sequences and their associated nuclear proteins. Finally, the saturation of the promoter in these transformed cell lines suggests that high levels of protein binding in the ODC promoter may contribute to elevated constitutive expression of this gene. Images PMID:1598217

  13. Characterization of the activity and expression of arginine decarboxylase in human and animal Chlamydia pathogens.

    PubMed

    Bliven, Kimberly A; Fisher, Derek J; Maurelli, Anthony T

    2012-12-01

    Chlamydia pneumoniae encodes a functional arginine decarboxylase (ArgDC), AaxB, that activates upon self-cleavage and converts l-arginine to agmatine. In contrast, most Chlamydia trachomatis serovars carry a missense or nonsense mutation in aaxB abrogating activity. The G115R missense mutation was not predicted to impact AaxB functionality, making it unclear whether AaxB variations in other Chlamydia species also result in enzyme inactivation. To address the impact of gene polymorphism on functionality, we investigated the activity and production of the Chlamydia AaxB variants. Because ArgDC plays a critical role in the Escherichia coli acid stress response, we studied the ability of these Chlamydia variants to complement an E. coli ArgDC mutant in an acid shock assay. Active AaxB was detected in four additional species: Chlamydia caviae, Chlamydia pecorum, Chlamydia psittaci, and Chlamydia muridarum. Of the C. trachomatis serovars, only E appears to encode active enzyme. To determine when functional enzyme is present during the chlamydial developmental cycle, we utilized an anti-AaxB antibody to detect both uncleaved and cleaved enzyme throughout infection. Uncleaved enzyme production peaked around 20 h postinfection, with optimal cleavage around 44 h. While the role ArgDC plays in Chlamydia survival or virulence is unclear, our data suggest a niche-specific function.

  14. Ornithine decarboxylase or gamma-glutamylcysteine synthetase overexpression protects Leishmania (Vianna) guyanensis against antimony.

    PubMed

    Fonseca, Maisa S; Comini, Marcelo A; Resende, Bethânia V; Santi, Ana Maria M; Zoboli, Antônio P; Moreira, Douglas S; Murta, Silvane M F

    2017-04-01

    Trypanosomatids present a unique mechanism for detoxification of peroxides that is dependent on trypanothione (bisglutathionylspermidine). Ornithine decarboxylase (ODC) and γ-glutamylcysteine synthetase (GSH1) produce molecules that are direct precursors of trypanothione. In this study, Leishmania guyanensis odc and gsh1 overexpressor cell lines were generated to investigate the contribution of these genes to the trivalent antimony (Sb(III))-resistance phenotype. The ODC- or GSH1-overexpressors parasites presented an increase of two and four-fold in Sb(III)-resistance index, respectively, when compared with the wild-type line. Pharmacological inhibition of ODC and GSH1 with the specific inhibitors α-difluoromethylornithine (DFMO) and buthionine sulfoximine (BSO), respectively, increased the antileishmanial effect of Sb(III) in all cell lines. However, the ODC- and GSH1-overexpressor were still more resistant to Sb(III) than the parental cell line. Together, our data shows that modulation of ODC and GSH1 levels and activity is sufficient to affect L. guyanensis susceptibility to Sb(III), and confirms a role of these genes in the Sb(III)-resistance phenotype.

  15. Effect of cocaine, ethanol or nicotine on ornithine decarboxylase activity in early chick embryo brain.

    PubMed

    Beeker, K; Smith, C; Pennington, S

    1992-09-18

    Fetal drug exposure causes multiple deficits in the developing child. For both humans and animal models, the single most common drug-related problem is fetal growth suppression. This defect is associated with significant perinatal morbidity and mortality and may also be related to significant behavioral problems appearing later in life. Studies focussed on the molecular mechanism of fetal drug effects in placental models are complicated by multiple interactions of the drug with mother, placenta and fetus. Using early (76-168 h) chick embryos as a non-placental model, and three common drugs of abuse (nicotine, ethanol and cocaine) it was found that each drug suppressed the peak in fetal brain ornithine decarboxylase (ODC) activity that normally occurs at 120 h of development. For each drug, the decrease in ODC activity at 120 h was followed by a small but significant increase in ODC. Thus, although the drug-treated embryos were smaller in size, they appeared to be undergoing compensatory growth and, in fact, became equal in weight to the vehicle-treated animals, if allowed to hatch.

  16. Substrate shuttling between active sites of uroporphyrinogen decarboxylase is not required to generate coproporphyrinogen

    PubMed Central

    Phillips, John D.; Warby, Christy A.; Whitby, Frank G.; Kushner, James P.; Hill, Christopher P.

    2009-01-01

    Summary Uroporphyrinogen Decarboxylase (URO-D; EC 4.1.1.37), the fifth enzyme of the heme biosynthetic pathway, is required for the production of heme, vitamin B12, siroheme, and chlorophyll precursors. URO-D catalyzes the sequential decarboxylation of the four acetate side chains on the pyrrole groups of uroporphyrinogen to produce coproporphyrinogen. URO-D is a stable homodimer with the active site clefts of the two subunits adjacent to each other. It has been hypothesized that the two catalytic centers interact functionally, perhaps by shuttling of reaction intermediates between subunits. We tested this hypothesis by construction of a single chain protein (scURO-D) in which the two subunits were connected by a flexible linker. The crystal structure of this protein was shown to be superimposible with wild-type activity and have comparable catalytic activity. Mutations that impaired one or the other of the two active sites of scURO-D resulted in approximately half of wild-type activity. The distribution of reaction intermediates was the same for mutant and wild-type sequences, and was unaltered in a competition experiment using the I and III isomer substrates. These observations indicate that communication between active sites is not required for enzyme function, and suggest that the dimeric structure of URO-D is required to achieve conformational stability and create a large active site cleft. PMID:19362562

  17. Coexpression of pyruvate decarboxylase and alcohol dehydrogenase genes in Lactobacillus brevis.

    PubMed

    Liu, Siqing; Dien, Bruce S; Nichols, Nancy N; Bischoff, Kenneth M; Hughes, Stephen R; Cotta, Michael A

    2007-09-01

    Lactobacillus brevis ATCC367 was engineered to express pyruvate decarboxylase (PDC) and alcohol dehydrogenase (ADH) genes in order to increase ethanol fermentation from biomass-derived residues. First, a Gram-positive Sarcina ventriculi PDC gene (Svpdc) was introduced into L. brevis ATCC 367 to obtain L. brevis bbc03. The SvPDC was detected by immunoblot using an SvPDC oligo peptide antiserum, but no increased ethanol was detected in L. brevis bbc03. Then, an ADH gene from L. brevis (Bradh) was cloned behind the Svpdc gene that generated a pdc/adh-coupled ethanol cassette pBBC04. The pBBC04 restored anaerobic growth and conferred ethanol production of Escheirichia coli NZN111 (a fermentative defective strain incapable of growing anaerobically). Approximately 58 kDa (SvPDC) and 28 kDa (BrADH) recombinant proteins were observed in L. brevis bbc04. These results indicated that the Gram-positive ethanol production genes can be expressed in L. brevis using a Gram-positive promoter and pTRKH2 shuttle vector. This work provides evidence that expressing Gram-positive ethanol genes in pentose utilizing L. brevis will further aid manipulation of this microbe toward biomass to ethanol production.

  18. Tyrosine decarboxylase activity of Lactobacillus brevis IOEB 9809 isolated from wine and L. brevis ATCC 367.

    PubMed

    Moreno-Arribas, V; Lonvaud-Funel, A

    1999-11-01

    Tyramine, a frequent amine in wines, is produced from tyrosine by the tyrosine decarboxylase (TDC) activity of bacteria. The tyramine-producing strain Lactobacillus brevis IOEB 9809 isolated from wine and the reference strain L. brevis ATCC 367 were studied. At the optimum pH, 5.0, K(m) values of IOEB 9809 and ATCC 367 crude extracts for L-tyrosine were 0.58 mM and 0.67 mM, and V(max) was higher for the wine strain (115 U) than the ATCC 367 (66 U). TDC exhibited a preference for L-tyrosine over L-DOPA as substrate. Enzyme activity was pyridoxal-5'-phosphate (PLP)-dependent and it was stabilized by the substrate and coenzyme. In contrast, glycerol and beta-mercaptoethanol strongly inhibited TDC. Tyramine competitively inhibited TDC for both strains. Citric acid, lactic acid and ethanol had an inhibitory effect on cells and crude extracts, but none could inhibit TDC at the usual concentrations in wines.

  19. Substrate Shuttling Between Active Sites of Uroporphyrinogen Decarboxylase in Not Required to Generate Coproporphyrinogen

    SciTech Connect

    Phillips, J.; Warby, C; Whitby, F; Kushner, J; Hill, C

    2009-01-01

    Uroporphyrinogen decarboxylase (URO-D; EC 4.1.1.37), the fifth enzyme of the heme biosynthetic pathway, is required for the production of heme, vitamin B12, siroheme, and chlorophyll precursors. URO-D catalyzes the sequential decarboxylation of four acetate side chains in the pyrrole groups of uroporphyrinogen to produce coproporphyrinogen. URO-D is a stable homodimer, with the active-site clefts of the two subunits adjacent to each other. It has been hypothesized that the two catalytic centers interact functionally, perhaps by shuttling of reaction intermediates between subunits. We tested this hypothesis by construction of a single-chain protein (single-chain URO-D) in which the two subunits were connected by a flexible linker. The crystal structure of this protein was shown to be superimposable with wild-type activity and to have comparable catalytic activity. Mutations that impaired one or the other of the two active sites of single-chain URO-D resulted in approximately half of wild-type activity. The distributions of reaction intermediates were the same for mutant and wild-type sequences and were unaltered in a competition experiment using I and III isomer substrates. These observations indicate that communication between active sites is not required for enzyme function and suggest that the dimeric structure of URO-D is required to achieve conformational stability and to create a large active-site cleft.

  20. Functional and conformational transitions of mevalonate diphosphate decarboxylase from Bacopa monniera.

    PubMed

    Abbassi, Shakeel; Patel, Krunal; Khan, Bashir; Bhosale, Siddharth; Gaikwad, Sushama

    2016-02-01

    Functional and conformational transitions of mevalonate diphosphate decarboxylase (MDD), a key enzyme of mevalonate pathway in isoprenoid biosynthesis, from Bacopa monniera (BmMDD), cloned and overexpressed in Escherichia coli were studied under thermal, chemical and pH-mediated denaturation conditions using fluorescence and Circular dichroism spectroscopy. Native BmMDD is a helix dominant structure with 45% helix and 11% sheets and possesses seven tryptophan residues with two residues exposed on surface, three residues partially exposed and two situated in the interior of the protein. Thermal denaturation of BmMDD causes rapid structural transitions at and above 40°C and transient exposure of hydrophobic residues at 50°C, leading to aggregation of the protein. An acid induced molten globule like structure was observed at pH 4, exhibiting altered but compact secondary structure, distorted tertiary structure and exposed hydrophobic residues. The molten globule displayed different response at higher temperature and similar response to chemical denaturation as compared to the native protein. The surface tryptophans have predominantly positively charged amino acids around them, as indicated by higher KSV for KI as compared to that for CsCl. The native enzyme displayed two different lifetimes, τ1 (1.203±0.036 ns) and τ2 (3.473±0.12 ns) indicating two populations of tryptophan.

  1. Partial purification and characterization of a novel histidine decarboxylase from Enterobacter aerogenes DL-1.

    PubMed

    Zou, Yu; Hu, Wenzhong; Jiang, Aili; Tian, Mixia

    2015-08-18

    Histidine decarboxylase (HDC) from Enterobacter aerogenes DL-1 was purified in a three-step procedure involving ammonium sulfate precipitation, Sephadex G-100, and DEAE-Sepharose column chromatography. The partially purified enzyme showed a single protein band of 52.4 kD on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The optimum pH for HDC activity was 6.5, and the enzyme was stable between pH 4 and 8. Enterobacter aerogenes HDC had optimal activity at 40°C and retained most of its activity between 4 and 50°C. HDC activity was reduced in the presence of numerous tested compounds. Particularly with SDS, it significantly (p < 0.01) inhibited enzyme activity. Conversely, Ca(2+) and Mn(2+) showed prominent activation effects (p < 0.01) with activity increasing to 117.20% and 123.42%, respectively. The Lineweaver-Burk plot showed that K m and V max values of the enzyme for L-histidine were 0.21 mM and 71.39 µmol/min, respectively. In comparison with most HDCs from other microorganisms and animals, HDC from E. aerogenes DL-1 displayed higher affinity and greater reaction velocity toward L-histidine.

  2. Induction of histidine decarboxylase in macrophages inhibited by the novel NF-{kappa}B inhibitor (-)-DHMEQ

    SciTech Connect

    Suzuki, Eriko Ninomiya, Yoko; Umezawa, Kazuo

    2009-02-06

    Histamine often causes inflammation, and this amine is produced by histidine decarboxylase (HDC). We found that (-)-DHMEQ, an NF-{kappa}B inhibitor, inhibited lipopolysaccharide (LPS)-induced histamine production and HDC induction in mouse macrophage cell line RAW264.7. However, as there is no {kappa}B site in the HDC promoter, we studied the mechanism of inhibition. Knockdown of the transcription factor C/EBP{beta} reduced the HDC expression in LPS-treated cells. (-)-DHMEQ inhibited the C/EBP{beta} transcriptional activity in a reporter assay and in an electrophoresis mobility shift assay. But it did not inhibit the in vitro binding of C/EBP{beta} to DNA. It also did not lower the nuclear amount of C/EBP{beta}. On the other hand, the addition of recombinant p65, a component of NF-{kappa}B, enhanced the activity of C/EBP{beta} acting as a cofactor in vitro. Then, we found that (-)-DHMEQ lowered the nuclear amount of p65. Thus, inhibition of the C/EBP{beta} activity by (-)-DHMEQ would be due to a reduction in the amount of nuclear p65, which has a co-activator activity for C/EBP{beta} that is essential for the HDC induction. (-)-DHMEQ may be useful as an anti-inflammatory agent by lowering the histamine production in the body.

  3. Renal ornithine decarboxylase activity, polyamines, and compensatory renal hypertrophy in the rat

    SciTech Connect

    Humphreys, M.H.; Etheredge, S.B.; Lin, Shanyan; Ribstein, J.; Marton, L.J. Univ. of California, San Francisco )

    1988-08-01

    The authors determined the role of ornithine decarboxylase (ODC) in compensatory renal hypertrophy (CRH) by relating renal ODC activity and polyamine content to kidney size, expressed as a percent of body weight, 1 wk after unilateral nephrectomy (UN). In normal rats, renal ODC activity increased after UN; 1 wk later the remaining kidney weight had increased. Renal concentration of putrescine, the product of ODC's decarboxylation of ornithine, was increased 3, 8, and 48 h after UN, but concentrations of polyamines synthesized later in the pathway, spermidine and spermine, were not appreciably affected. Pretreatment with difluoromethylornithine (DFMO), an irreversible inhibitor of ODC inhibited both base-line renal ODC activity and putrescine concentration as well as increases stimulated by UN, although concentrations of spermidine and spermine were not decreased. In hypophysectomized rats, both increased renal ODC activity and CRH occurred as well, indicating that these two consequences of UN do not require intact pituitary function. Thus stimulation of renal ODC activity and putrescine content do not appear critical to the process of CRH after UN.

  4. Aspartate beta-decarboxylase from Alcaligenes faecalis: carbon-13 kinetic isotope effect and deuterium exchange experiments

    SciTech Connect

    Rosenberg, R.M.; O'Leary, M.H.

    1985-03-26

    The authors have measured the /sup 13/C kinetic isotope effect at pH 4.0, 5.0, 6.0, and 6.5 and in D/sub 2/O at pH 5.0 and the rate of D-H exchange of the alpha and beta protons of aspartic acid in D/sub 2/O at pH 5.0 for the reaction catalyzed by the enzyme aspartate beta-decarboxylase from Alcaligenes faecalis. The /sup 13/C kinetic isotope effect, with a value of 1.0099 +/- 0.0002 at pH 5.0, is less than the intrinsic isotope effect for the decarboxylation step, indicating that the decarboxylation step is not entirely rate limiting. The authors have been able to estimate probable values of the relative free energies of the transition states of the enzymatic reaction up to and including the decarboxylation step from the /sup 13/C kinetic isotope effect and the rate of D-H exchange of alpha-H. The pH dependence of the kinetic isotope effect reflects the pKa of the pyridine nitrogen of the coenzyme pyridoxal 5'-phosphate but not that of the imine nitrogen. A mechanism is proposed for the exchange of aspartate beta-H that is consistent with the stereochemistry suggested earlier.

  5. Identification and characterization of barley mutants lacking glycine decarboxylase and carboxyl esterase activities

    SciTech Connect

    Blackwell, R.; Lewis, K.; Lea, P. )

    1990-05-01

    A barley mutant has been isolated, from a selection of fifty air-sensitive seed-lines, using a standard gel stain technique which lacks carboxyl esterase activity, but has normal levels of carbonic anhydrase. In addition, two barley mutants lacking the ability to convert glycine to serine in the mitochondria, have been characterized. Both plants accumulate glycine in air and are unable to metabolize ({sup 14}C)glycine in the short-term. When ({sup 14}C)glycine was supplied over 2h LaPr 85/55 metabolized 90%, whereas the second mutant (LaPr 87/30) metabolized 10%. Results indicate that the mutation in LaPr 85/55 is almost certainly in the glycine transporter into the mitochondrion. The mutation in LaPr 87/30 has been shown, using western blotting, to be in both the P and H proteins, two of four proteins which comprise glycine decarboxylase (P, H, T and L).

  6. Induction of an oxalate decarboxylase in the filamentous fungus Trametes versicolor by addition of inorganic acids.

    PubMed

    Zhu, Cui Xia; Hong, Feng

    2010-01-01

    In order to improve yields and to reduce the cost of oxalate decarboxylase (OxDC, EC 4.1.1.2), the induction of OxDC in the white-rot fungus Trametes versicolor was studied in this work. OxDC was induced by addition of inorganic acids including hydrochloric acid, sulfuric acid, and phosphoric acid to culture media. The results showed that all the acids could enhance OxDC expression. The activity of the acid-induced OxDC rose continuously. All of the OxDC volumetric activities induced by the inorganic acids were higher than 20.0 U/L and were two times higher than that obtained with oxalic acid. OxDC productivity was around 4.0 U*L(-1)*day(-1). The highest specific activity against total protein was 3.2 U/mg protein at day 8 after induction of sulfuric acid, and the specific activity against mycelial dry weight was 10.6 U/g at day 9 after induction of hydrochloric acid. The growth of mycelia was inhibited slightly when the pH values in culture media was around 2.5-3.0, while the growth was inhibited heavily when the pH was lower than 2.5.

  7. A high-throughput colorimetric assay to measure the activity of glutamate decarboxylase.

    PubMed

    Yu, Kai; Hu, Sheng; Huang, Jun; Mei, Le-He

    2011-08-10

    A pH-sensitive colorimetric assay has been established to quantitatively measure glutamate decarboxylase (GAD) activity in bacterial cell extracts using a microplate format. GAD catalyzes the irreversible α-decarboxylation of L-glutamate to γ-aminobutyrate. The assay is based on the color change of bromocresol green due to an increase in pH as protons are consumed during the enzyme-catalyzed reaction. Bromocresol green was chosen as the indicator because it has a similar pK(a) to the acetate buffer used. The corresponding absorbance change at 620 nm was recorded with a microplate reader as the reaction proceeded. A difference in the enzyme preparation pH and optimal pH for GAD activity of 2.5 did not prevent this method from successfully allowing the determination of reaction kinetic parameters and the detection of improvements in enzymatic activity with a low coefficient of variance. Our assay is simple, rapid, requires minimal sample concentration and can be carried out in robotic high-throughput devices used as standard in directed evolution experiments. In addition, it is also applicable to other reactions that involve a change in pH.

  8. Increasing thermal stability and catalytic activity of glutamate decarboxylase in E. coli: An in silico study.

    PubMed

    Tavakoli, Yasaman; Esmaeili, Abolghasem; Saber, Hossein

    2016-10-01

    Glutamate decarboxylase (GAD) is an enzyme that converts l-glutamate to gamma amino butyric acid (GABA) that is a widely used drug to treat mental disorders like Alzheimer's disease. In this study for the first time point mutation was performed virtually in the active site of the E. coli GAD in order to increase thermal stability and catalytic activity of the enzyme. Energy minimization and addition of water box were performed using GROMACS 5.4.6 package. PoPMuSiC 2.1 web server was used to predict potential spots for point mutation and Modeller software was used to perform point mutation on three dimensional model. Molegro virtual docker software was used for cavity detection and stimulated docking study. Results indicate that performing mutation separately at positions 164, 302, 304, 393, 396, 398 and 410 increase binding affinity to substrate. The enzyme is predicted to be more thermo- stable in all 7 mutants based on ΔΔG value.

  9. Complexes of Thermotoga maritima S-adenosylmethionine decarboxylase provide insights into substrate specificity

    SciTech Connect

    Bale, Shridhar; Baba, Kavita; McCloskey, Diane E.; Pegg, Anthony E.; Ealick, Steven E.

    2010-06-25

    The polyamines putrescine, spermidine and spermine are ubiquitous aliphatic cations and are essential for cellular growth and differentiation. S-Adenosylmethionine decarboxylase (AdoMetDC) is a critical pyruvoyl-dependent enzyme in the polyamine-biosynthetic pathway. The crystal structures of AdoMetDC from humans and plants and of the AdoMetDC proenzyme from Thermotoga maritima have been obtained previously. Here, the crystal structures of activated T. maritima AdoMetDC (TmAdoMetDC) and of its complexes with S-adenosylmethionine methyl ester and 5{prime}-deoxy-5{prime}-dimethylthioadenosine are reported. The results demonstrate for the first time that TmAdoMetDC autoprocesses without the need for additional factors and that the enzyme contains two complete active sites, both of which use residues from both chains of the homodimer. The complexes provide insights into the substrate specificity and ligand binding of AdoMetDC in prokaryotes. The conservation of the ligand-binding mode and the active-site residues between human and T. maritima AdoMetDC provides insight into the evolution of AdoMetDC.

  10. [Enhancing glutamate decarboxylase activity by site-directed mutagenesis: an insight from Ramachandran plot].

    PubMed

    Ke, Piyu; Huang, Jun; Hu, Sheng; Zhao, Weirui; Lü, Changjiang; Yu, Kai; Lei, Yinlin; Wang, Jinbo; Mei, Lehe

    2016-01-01

    Glutamate decarboxylase (GAD) can catalyze the decarboxylation of glutamate into γ-aminobutyrate (GABA) and is the only enzyme of GABA biosynthesis. Improving GAD activity and thermostability will be helpful for the highly efficient biosynthesis of GABA. According to the Ramachandran plot information of GAD 1407 three-dimensional structure from Lactobacillus brevis CGMCC No. 1306, we identified the unstable site K413 as the mutation target, constructed the mutant GAD by site-directed mutagenesis and measured the thermostability and activity of the wide type and mutant GAD. Mutant K413A led to a remarkably slower inactivation rate, and its half-life at 50 °C reached 105 min which was 2.1-fold higher than the wild type GAD1407. Moreover, mutant K413I exhibited 1.6-fold higher activity in comparison with the wide type GAD1407, although it had little improvement in thermostability of GAD. Ramachandran plot can be considered as a potential approach to increase GAD thermostability and activity.

  11. Quantitative expression analysis and prognostic significance of L-DOPA decarboxylase in colorectal adenocarcinoma

    PubMed Central

    Kontos, C K; Papadopoulos, I N; Fragoulis, E G; Scorilas, A

    2010-01-01

    Background: L-DOPA decarboxylase (DDC) is an enzyme that catalyses, mainly, the decarboxylation of L-DOPA to dopamine and was found to be involved in many malignancies. The aim of this study was to investigate the mRNA expression levels of the DDC gene and to evaluate its clinical utility in tissues with colorectal adenocarcinoma. Methods: Total RNA was isolated from colorectal adenocarcinoma tissues of 95 patients. After having tested RNA quality, we prepared cDNA by reverse transcription. Highly sensitive quantitative real-time PCR method for DDC mRNA quantification was developed using the SYBR Green chemistry. GAPDH served as a housekeeping gene. Relative quantification analysis was performed using the comparative CT method (2−ΔΔCT). Results: DDC mRNA expression varied remarkably among colorectal tumours examined in this study. High DDC mRNA expression levels were found in well-differentiated and Dukes' stage A and B tumours. Kaplan–Meier survival curves showed that patients with DDC-positive tumours have significantly longer disease-free survival (P=0.009) and overall survival (P=0.027). In Cox regression analysis of the entire cohort of patients, negative DDC proved to be a significant predictor of reduced disease-free (P=0.021) and overall survival (P=0.047). Conclusions: The results of the study suggest that DDC mRNA expression may be regarded as a novel potential tissue biomarker in colorectal adenocarcinoma. PMID:20424616

  12. Elevated production of melatonin in transgenic rice seeds expressing rice tryptophan decarboxylase.

    PubMed

    Byeon, Yeong; Park, Sangkyu; Lee, Hyoung Yool; Kim, Young-Soon; Back, Kyoungwhan

    2014-04-01

    A major goal of plant biotechnology is to improve the nutritional qualities of crop plants through metabolic engineering. Melatonin is a well-known bioactive molecule with an array of health-promoting properties, including potent antioxidant capability. To generate melatonin-rich rice plants, we first independently overexpressed three tryptophan decarboxylase isogenes in the rice genome. Melatonin levels were altered in the transgenic lines through overexpression of TDC1, TDC2, and TDC3; TDC3 transgenic seed (TDC3-1) had melatonin concentrations 31-fold higher than those of wild-type seeds. In TDC3 transgenic seedlings, however, only a doubling of melatonin content occurred over wild-type levels. Thus, a seed-specific accumulation of melatonin appears to occur in TDC3 transgenic lines. In addition to increased melatonin content, TDC3 transgenic lines also had enhanced levels of melatonin intermediates including 5-hydroxytryptophan, tryptamine, serotonin, and N-acetylserotonin. In contrast, expression levels of melatonin biosynthetic mRNA did not increase in TDC3 transgenic lines, indicating that increases in melatonin and its intermediates in these lines are attributable exclusively to overexpression of the TDC3 gene.

  13. S-adenosyl-L-methionine decarboxylase activity in the rat epididymis: ontogeny and androgenic control.

    PubMed

    de las Heras, M A; Calandra, R S

    1991-01-01

    The authors describe the occurrence of high levels of S-adenosyl-L-methionine decarboxylase (SAMDC) activity in the rat epididymis, and its ontogeny and androgenic control. As early as 15 days of age, SAMDC activity exists, although a peak of activity is observed at 25 days. Bilateral orchidectomy resulted in a decline of epididymal SAMDC activity. However, an androgen-independent fraction, accounting for 34% of total activity, appears to exist in the epididymis. In 45-day-old orchidectomized rats, SAMDC activity was stimulated by testosterone treatment in a dose-dependent manner. However, treatment of 45-day-old intact animals with a high dose of the androgen failed to modify SAMDC activity, indicating that, at this age, the enzyme is maximally stimulated by endogenous androgens. The observed effect of testosterone on castrated rats was completely abolished by concomitant treatment with the antiandrogen flutamide. This compound was ineffective on the androgen-insensitive fraction. To assess the contribution of circulating and luminal androgens to the maintenance of epididymal SAMDC, rats were unilaterally orchidectomized and activity was determined in both epididymides after 7 days. The SAMDC activity was identical in epididymides from both sides, suggesting circulating androgens suffice to maintain normal levels of activity. It was concluded that androgens regulate epididymal SAMDC activity, although an androgen-independent fraction appears to exist.

  14. Acute effect of prolactin on ornithine decarboxylase activity in the rat testis.

    PubMed

    de Las Heras, M A; Calandra, R S

    1992-01-01

    A study was conducted to evaluate the effect of the acute treatment with prolactin (PRL) on ornithine decarboxylase (ODC) activity in the rat testis. Injection of a single SC dose of ovine PRL to puberal rats resulted in the activation of ODC from whole testis. This effect was maximal at 4 h after injection, and statistically significant at the dose of 500 micrograms. The effect of PRL was confined to the interstitial space; no change was observed in seminiferous tubules. PRL was unable to further increase testicular ODC activity when injected together with a stimulatory dose of human chorionic gonadotropin (hCG). The effect of PRL was mimicked by injection of a single dose of the dopamine antagonist sulpiride, which provoked a ninefold increase in serum PRL levels. In contrast, PRL did not stimulate testicular ODC activity in hypophysectomized rats, either under basal conditions or during treatment with PRL-hCG, indicating the requirement of a functional hypophysis for the expression of PRL action. These results suggest that the stimulation of testicular ODC activity by PRL is a marker of the trophic response of the testis to this hormone, different from the stimulation of steroidogenesis. This activity could be useful for the study of PRL action on the testis as well as of the interaction between PRL and LH at the testicular level.

  15. The bifunctional pyruvate decarboxylase/pyruvate ferredoxin oxidoreductase from Thermococcus guaymasensis.

    PubMed

    Eram, Mohammad S; Oduaran, Erica; Ma, Kesen

    2014-01-01

    The hyperthermophilic archaeon Thermococcus guaymasensis produces ethanol as a metabolic end product, and an alcohol dehydrogenase (ADH) catalyzing the reduction of acetaldehyde to ethanol has been purified and characterized. However, the enzyme catalyzing the formation of acetaldehyde has not been identified. In this study an enzyme catalyzing the production of acetaldehyde from pyruvate was purified and characterized from T. guaymasensis under strictly anaerobic conditions. The enzyme had both pyruvate decarboxylase (PDC) and pyruvate ferredoxin oxidoreductase (POR) activities. It was oxygen sensitive, and the optimal temperatures were 85°C and >95°C for the PDC and POR activities, respectively. The purified enzyme had activities of 3.8 ± 0.22 U mg(-1) and 20.2 ± 1.8 U mg(-1), with optimal pH-values of 9.5 and 8.4 for each activity, respectively. Coenzyme A was essential for both activities, although it did not serve as a substrate for the former. Enzyme kinetic parameters were determined separately for each activity. The purified enzyme was a heterotetramer. The sequences of the genes encoding the subunits of the bifunctional PDC/POR were determined. It is predicted that all hyperthermophilic β -keto acids ferredoxin oxidoreductases are bifunctional, catalyzing the activities of nonoxidative and oxidative decarboxylation of the corresponding β -keto acids.

  16. Differential induction of pyruvate decarboxylase subunits and transcripts in anoxic rice seedlings.

    PubMed Central

    Rivoal, J; Thind, S; Pradet, A; Ricard, B

    1997-01-01

    In 2-d-old rice (Oryza sativa L.) seedlings subjected to anoxic stress, pyruvate decarboxylase (PDC) activity increased 9-fold during a 168-h period. A polyclonal PDC antiserum that recognized alpha- and beta-subunits was used to quantify PDC protein by an enzyme-linked immunosorbant assay and showed a 5.6-fold increase, suggesting that the anoxically induced enzyme has a higher specific activity than the PDC isoform present under normoxia. Immunoblot analysis showed that levels of both PDC subunits were induced by anoxia. Immunoprecipitation of proteins labeled in vivo during anoxic treatment demonstrated that the alpha-subunit was preferentially synthesized at the onset of anoxia. Two partial cDNAs, including a novel sequence, were cloned from a cDNA library made from seedlings subjected to anoxia for 6 h. Gene-specific probes used to quantify northern blots showed that two or three PDC mRNAs are differentially induced by anoxia in rice seedlings. Immunoprecipitation of in vitro translation products of mRNAs isolated a different times of anoxic treatment confirmed this findings Our results suggest that anoxic induction of rice PDC involves transcriptional and posttranscriptional regulation of gene expression as well as differences in enzyme characteristics. PMID:9232881

  17. Partial purification and characterization of arginine decarboxylase from avocado fruit, a thermostable enzyme.

    PubMed

    Winer, L; Vinkler, C; Apelbaum, A

    1984-09-01

    A partially purified preparation of arginine decarboxylase (EC 4.1.1.19), a key enzyme in polyamine metabolism in plants, was isolated from avocado (Persea americana Mill. cv Fuerte) fruit. The preparation obtained from the crude extract after ammonium sulfate precipitation, dialysis, and heat treatment, had maximal activity between pH 8.0 and 9.0 at 60 degrees C, in the presence of 1.2 millimolar MnCl(2), 2 millimolar dithiothreitol, and 0.06 millimolar pyridoxal phosphate. The K(m), of arginine for the decarboxylation reaction was determined for enzymes prepared from the seed coat of both 4-week-old avocado fruitlet and fully developed fruit, and was found to have a value of 1.85 and 2.84 millimolar, respectively. The value of V(app) (max) of these enzymes was 1613 and 68 nanomoles of CO(2) produced per milligram of protein per hour for the fruitlet and the fully developed fruit, respectively. Spermine, an end product of polyamine metabolism, caused less than 5% inhibition of the enzyme from fully developed fruit and 65% inhibition of the enzyme from the seed coat of 4-week-old fruitlets at 1 millimolar under similar conditions. The effect of different inhibitors on the enzyme and the change in the nature of the enzyme during fruit development are discussed.

  18. Biochemical and Genetic Characterization of the Enterococcus faecalis Oxaloacetate Decarboxylase Complex

    PubMed Central

    Repizo, Guillermo D.; Blancato, Víctor S.; Mortera, Pablo; Lolkema, Juke S.

    2013-01-01

    Enterococcus faecalis encodes a biotin-dependent oxaloacetate decarboxylase (OAD), which is constituted by four subunits: E. faecalis carboxyltransferase subunit OadA (termed Ef-A), membrane pump Ef-B, biotin acceptor protein Ef-D, and the novel subunit Ef-H. Our results show that in E. faecalis, subunits Ef-A, Ef-D, and Ef-H form a cytoplasmic soluble complex (termed Ef-AHD) which is also associated with the membrane. In order to characterize the role of the novel Ef-H subunit, coexpression of oad genes was performed in Escherichia coli, showing that this subunit is vital for Ef-A and Ef-D interaction. Diminished growth of the oadA and oadD single deletion mutants in citrate-supplemented medium indicated that the activity of the complex is essential for citrate utilization. Remarkably, the oadB-deficient strain was still capable of growing to wild-type levels but with a delay during the citrate-consuming phase, suggesting that the soluble Ef-AHD complex is functional in E. faecalis. These results suggest that the Ef-AHD complex is active in its soluble form, and that it is capable of interacting in a dynamic way with the membrane-bound Ef-B subunit to achieve its maximal alkalinization capacity during citrate fermentation. PMID:23435880

  19. pH shift enhancement of Candida utilis pyruvate decarboxylase production.

    PubMed

    Chen, Allen Kuan-Liang; Breuer, Michael; Hauer, Bernhard; Rogers, Peter L; Rosche, Bettina

    2005-10-20

    Pyruvate decarboxylase (PDC) catalyses the synthesis of asymmetric carbinols, e.g., chiral precursors for pharmaceuticals such as ephedrine and pseudoephedrine. The production of PDC by Candida utilis in a minimal medium was improved by manipulating the pH during fermentation in a 5 L bioreactor. At an aeration rate of 0.1 vvm with a stirrer speed of 300 rpm at constant pH 6, a specific PDC activity of 141 U/g dry cell weight (DCW) was achieved (average of two fermentations +/-13%). By allowing the yeast to acidify the growth medium from pH 6 to 2.9, the final specific PDC activity increased by a factor of 2.7 to 385 U/g DCW (average from 4 fermentations +/-16%). The effect of this pH drift on PDC production was confirmed by another experiment with a manual shift of pH from 6 to 3 by addition of 5 M sulfuric acid. The final PDC activity was 392 U/g DCW (average from two fermentations +/-5%). However, experiments with constant pH of 6, 5, 4, or 3 resulted in average specific activities of only 102 to 141 U/g DCW, suggesting that a transitional pH change rather than the absolute pH value was responsible for the increased specific PDC activity.

  20. Effect of undernutrition on the regional development of transmitter enzymes: glutamate decarboxylase and choline acetyltransferase.

    PubMed

    Patel, A J; del Vecchio, M; Atkinson, D J

    1978-01-01

    The effect of undernutrition on the activity of glutamate decarboxylase (GAD) and choline acetyltransferase (ChAc) (markers for the GABA-ergic and the cholinergic transmitter system, respectively) was studied in various parts of the rat brain at the age of 10, 15 and 21 days, and at day 54 following 33 days of rehabilitation. The brain regions investigated were the olfactory bulbs, cerebellum, pons-medulla, hypothalamus, colliculi, cerebral cortex hippocampus and the residual brain. Undernutrition resulted in a marked retardation of the developmental rise of the activities of both enzymes, expressed in terms of either total brain part or unit weight or protein. The effect diminished with age even during the period of nutritional deprivation. In most brain regions the enzyme activities were restored to normal after rehabilitation. In the cerebral cortex the total activity of both enzymes was persistently reduced, although the concentration of GAD exceeded the control levels. A negative correlation was manifested between the activities of GAD and ChAc in the different brain parts (except the cerebellum) during development. The correlation became significant by day 21 in the controls, but only after postweaning rehabilitation of the undernourished rats. The results showed therefore that undernutrition caused a reversible retardation in the development of these two transmitter enzymes, and they suggested that even the balance of the GABA-ergic and cholinergic systems throughout the brain can be restored to normal by rehabilitation.

  1. Polyamine metabolism and osmotic stress. II. Improvement of oat protoplasts by an inhibitor of arginine decarboxylase

    NASA Technical Reports Server (NTRS)

    Tiburcio, A. F.; Kaur-Sawhney, R.; Galston, A. W.

    1986-01-01

    We have attempted to improve the viability of cereal mesophyll protoplasts by pretreatment of leaves with DL-alpha-difluoromethylarginine (DFMA), a specific 'suicide' inhibitor of the enzyme (arginine decarboxylase) responsible for their osmotically induced putrescine accumulation. Leaf pretreatment with DFMA before a 6 hour osmotic shock caused a 45% decrease of putrescine and a 2-fold increase of spermine titer. After 136 hours of osmotic stress, putrescine titer in DFMA-pretreated leaves increased by only 50%, but spermidine and spermine titers increased dramatically by 3.2- and 6-fold, respectively. These increases in higher polyamines could account for the reduced chlorophyll loss and enhanced ability of pretreated leaves to incorporate tritiated thymidine, uridine, and leucine into macromolecules. Pretreatment with DFMA significantly improved the overall viability of the protoplasts isolated from these leaves. The results support the view that the osmotically induced rise in putrescine and blockage of its conversion to higher polyamines may contribute to the lack of sustained cell division in cereal mesophyll protoplasts, although other undefined factors must also play a major role.

  2. Ornithine decarboxylase and extracellular polyamines regulate microvascular sprouting and actin cytoskeleton dynamics in endothelial cells

    SciTech Connect

    Kucharzewska, Paulina; Welch, Johanna E.; Svensson, Katrin J.; Belting, Mattias

    2010-10-01

    The polyamines are essential for cancer cell proliferation during tumorigenesis. Targeted inhibition of ornithine decarboxylase (ODC), i.e. a key enzyme of polyamine biosynthesis, by {alpha}-difluoromethylornithine (DFMO) has shown anti-neoplastic activity in various experimental models. This activity has mainly been attributed to the anti-proliferative effect of DFMO in cancer cells. Here, we provide evidence that unperturbed ODC activity is a requirement for proper microvessel sprouting ex vivo as well as the migration of primary human endothelial cells. DFMO-mediated ODC inhibition was reversed by extracellular polyamine supplementation, showing that anti-angiogenic effects of DFMO were specifically related to polyamine levels. ODC inhibition was associated with an abnormal morphology of the actin cytoskeleton during cell spreading and migration. Moreover, our data suggest that de-regulated actin cytoskeleton dynamics in DFMO treated endothelial cells may be related to constitutive activation of the small GTPase CDC42, i.e. a well-known regulator of cell motility and actin cytoskeleton remodeling. These insights into the potential role of polyamines in angiogenesis should stimulate further studies testing the combined anti-tumor effect of polyamine inhibition and established anti-angiogenic therapies in vivo.

  3. Expanding the active pH range of Escherichia coli glutamate decarboxylase by breaking the cooperativeness.

    PubMed

    Thu Ho, Ngoc Anh; Hou, Chen Yuan; Kim, Woo Hyun; Kang, Taek Jin

    2013-02-01

    Bacterial glutamate decarboxylase (GAD) transforms glutamate into γ-aminobutyric acid (GABA) with the consumption of a proton. The enzyme is active under acidic environments only and sharply loses its activity as pH approaches neutrality with concomitant structural deformation. In an attempt to understand better the role of this cooperative loss of activity upon pH shifts, we prepared and studied a series of GAD site-specific mutants. In this report, we show that the cooperativeness was kept intact by at least two residues, Glu89 and His465, of which Glu89 is newly identified to be involved in the cooperativity system of GAD. Double mutation on these residues not only broke the cooperativity in the activity change but also yielded a mutant GAD that retained the activity at neutral pH. The resulting mutant GAD that was active at neutral pH inhibited the cell growth in a glycerol medium by converting intracellular Glu into GABA in an uncontrolled manner, which explains in part why the cooperativeness of GAD has to be kept by several layers of safety keepers. This unexpected result might be utilized to convert a low-valued by-product of biodiesel production, glycerol, into value-added product, GABA.

  4. Hepatoerythropoietic Porphyria Caused by a Novel Homoallelic Mutation in Uroporphyrinogen Decarboxylase Gene in Egyptian Patients.

    PubMed

    Farrag, M S; Mikula, I; Richard, E; Saudek, V; De Verneuil, H; Martásek, P

    2015-01-01

    Porphyrias are metabolic disorders resulting from mutations in haem biosynthetic pathway genes. Hepatoerythropoietic porphyria (HEP) is a rare type of porphyria caused by the deficiency of the fifth enzyme (uroporphyrinogen decarboxylase, UROD) in this pathway. The defect in the enzymatic activity is due to biallelic mutations in the UROD gene. Currently, 109 UROD mutations are known. The human disease has an early onset, manifesting in infancy or early childhood with red urine, skin photosensitivity in sun-exposed areas, and hypertrichosis. Similar defects and links to photosensitivity and hepatopathy exist in several animal models, including zebrafish and mice. In the present study, we report a new mutation in the UROD gene in Egyptian patients with HEP. We show that the homozygous c.T163A missense mutation leads to a substitution of a conserved phenylalanine (amino acid 55) for isoleucine in the enzyme active site, causing a dramatic decrease in the enzyme activity (19 % of activity of wild-type enzyme). Inspection of the UROD crystal structure shows that Phe-55 contacts the substrate and is located in the loop that connects helices 2 and 3. Phe-55 is strictly conserved in both prokaryotic and eukaryotic UROD. The F55I substitution likely interferes with the enzyme-substrate interaction.

  5. Dynamic expression of a glutamate decarboxylase gene in multiple non-neural tissues during mouse development

    PubMed Central

    Maddox, Dennis M; Condie, Brian G

    2001-01-01

    Background Glutamate decarboxylase (GAD) is the biosynthetic enzyme for the neurotransmitter γ-aminobutyric acid (GABA). Mouse embryos lacking the 67-kDa isoform of GAD (encoded by the Gad1 gene) develop a complete cleft of the secondary palate. This phenotype suggests that this gene may be involved in the normal development of tissues outside of the CNS. Although Gad1 expression in adult non-CNS tissues has been noted previously, no systematic analysis of its embryonic expression outside of the nervous system has been performed. The objective of this study was to define additional structures outside of the central nervous system that express Gad1, indicating those structures that may require its function for normal development. Results Our analysis detected the localized expression of Gad1 transcripts in several developing tissues in the mouse embryo from E9.0-E14.5. Tissues expressing Gad1 included the tail bud mesenchyme, the pharyngeal pouches and arches, the ectodermal placodes of the developing vibrissae, and the apical ectodermal ridge (AER), mesenchyme and ectoderm of the limb buds. Conclusions Some of the sites of Gad1 expression are tissues that emit signals required for patterning and differentiation (AER, vibrissal placodes). Other sites correspond to proliferating stem cell populations that give rise to multiple differentiated tissues (tail bud mesenchyme, pharyngeal endoderm and mesenchyme). The dynamic expression of Gad1 in such tissues suggests a wider role for GABA signaling in development than was previously appreciated. PMID:11178105

  6. Novel interactions of fluorinated nucleotide derivatives targeting orotidine-5′-monophosphate decarboxylase

    PubMed Central

    Lewis, Melissa; Avina, Maria Elena Meza; Wei, Lianhu; Crandall, Ian E.; Bello, Angelica Mara; Poduch, Ewa; Liu, Yan; Paige, Christopher J.; Kain, Kevin C.; Pai, Emil F.; Kotra, Lakshmi P.

    2011-01-01

    Fluorinated nucleosides and nucleotides are of considerable interest to medicinal chemists due to their antiviral, anticancer, and other biological activities. However, their direct interactions at target binding sites are not well understood. A new class of 2′-deoxy-2′-fluoro-C6-substituted uridine and UMP derivatives were synthesized and evaluated as inhibitors of orotidine-5′-monophosphate decarboxylase (ODCase). These compounds were synthesized from the key intermediate, fully-protected 2′-deoxy-2′-fluorouridine. Among the synthesized compounds, 2′-deoxy-2′-fluoro-6-iodo-UMP covalently inhibited human ODCase with a second-order rate constant of 0.62 ± 0.02 M−1sec−1. Interestingly, the 6-cyano-2′-fluoro derivative covalently interacted with ODCase defying the conventional thinking, where its ribosyl derivative undergoes transformation into BMP by ODCase. This confirms that the 2′-fluoro moiety influences the chemistry at the C6 position of the nucleotides, thus interactions in the active site of ODCase. Molecular interactions of the 2′-fluorinated nucleotides are compared to those with the 3′-fluorinated nucleotides bound to the corresponding target enzyme, and the carbohydrate moieties were shown to bind in different conformations. PMID:21417464

  7. Gene cloning, expression, and characterization of phenolic acid decarboxylase from Lactobacillus brevis RM84.

    PubMed

    Landete, José María; Rodríguez, Héctor; Curiel, José Antonio; de las Rivas, Blanca; Mancheño, José Miguel; Muñoz, Rosario

    2010-06-01

    Phenolic acid decarboxylase (PAD) catalyzes the synthesis of vinyl phenols from hydroxycinnamic acids. The gene encoding PAD from Lactobacillus brevis was cloned and expressed as a fusion protein in Escherichia coli. The recombinant PAD enzyme is a heat-labile enzyme that functions optimally at 22 degrees C and pH 6.0. The purified enzyme did not show thermostability at temperatures above 22 degrees C. L. brevis PAD is able to decarboxylate exclusively the hydroxycinnamic acids, such as p-coumaric, caffeic, and ferulic acids, with K (m) values of 0.98, 0.96, and 0.78 mM, respectively. The substrate specificity exhibited by L. brevis PAD is similar to the PAD isolated from Bacillus subtilis and B. pumilus, but different from that of L. plantarum and Pediococcus pentosaceus. As the C-terminal region may be involved in determining PAD substrate specificity and catalytic capacity, amino acid differences among these proteins could explain the differences observed. The substrate specificity shown by L. brevis PAD shows promise for the synthesis of high-added value products from plant wastes.

  8. Glycine decarboxylase deficiency causes neural tube defects and features of non-ketotic hyperglycinemia in mice

    PubMed Central

    Pai, Yun Jin; Leung, Kit-Yi; Savery, Dawn; Hutchin, Tim; Prunty, Helen; Heales, Simon; Brosnan, Margaret E.; Brosnan, John T.; Copp, Andrew J.; Greene, Nicholas D.E.

    2015-01-01

    Glycine decarboxylase (GLDC) acts in the glycine cleavage system to decarboxylate glycine and transfer a one-carbon unit into folate one-carbon metabolism. GLDC mutations cause a rare recessive disease non-ketotic hyperglycinemia (NKH). Mutations have also been identified in patients with neural tube defects (NTDs); however, the relationship between NKH and NTDs is unclear. We show that reduced expression of Gldc in mice suppresses glycine cleavage system activity and causes two distinct disease phenotypes. Mutant embryos develop partially penetrant NTDs while surviving mice exhibit post-natal features of NKH including glycine accumulation, early lethality and hydrocephalus. In addition to elevated glycine, Gldc disruption also results in abnormal tissue folate profiles, with depletion of one-carbon-carrying folates, as well as growth retardation and reduced cellular proliferation. Formate treatment normalizes the folate profile, restores embryonic growth and prevents NTDs, suggesting that Gldc deficiency causes NTDs through limiting supply of one-carbon units from mitochondrial folate metabolism. PMID:25736695

  9. Overexpression of Tyrosine hydroxylase and Dopa decarboxylase associated with pupal melanization in Spodoptera exigua

    PubMed Central

    Liu, Sisi; Wang, Mo; Li, Xianchun

    2015-01-01

    Melanism has been found in a wide range of species, but the molecular mechanisms involved remain largely elusive. In this study, we studied the molecular mechanisms of the pupal melanism in Spodoptera exigua. The full length cDNA sequences of tyrosine hydroxylase (TH) and dopa decarboxylase (DDC), two key enzymes in the biosynthesis pathway of melanin, were cloned, and their temporal expression patterns in the integument were compared during the larval-pupal metamorphosis process of the S. exigua wild type (SEW) and melanic mutant (SEM) strains. No amino acid change in the protein sequence of TH and DDC was found between the two strains. Both DDC and TH were significantly over-expressed in the integument of the SEM strain at late-prepupa and 0 h pupa, respectively, compared with those of the SEW strain. Feeding 5th instar larvae of SEM with diets incorporated with 1 mg/g of the DDC inhibitor L-α-Methyl-DOPA and 0.75 mg/g of the TH inhibitor 3-iodo-tyrosine (3-IT) resulted in 20% pupae with partially-rescued phenotype and 68.2% of pupae with partially- or fully-rescued phenotype, respectively. These results indicate that overexpressions of TH and DDC are involved in the pupal melanization of S. exigua. PMID:26084938

  10. Polyamines directly promote antizyme-mediated degradation of ornithine decarboxylase by the proteasome

    PubMed Central

    Beenukumar, R. R.; Gödderz, Daniela; Palanimurugan, R.; Dohmen, R. J.

    2015-01-01

    Ornithine decarboxylase (ODC), a ubiquitin-independent substrate of the proteasome, is a homodimeric protein with a rate-limiting function in polyamine biosynthesis. Polyamines regulate ODC levels by a feedback mechanism mediated by ODC antizyme (OAZ). Higher cellular polyamine levels trigger the synthesis of OAZ and also inhibit its ubiquitin-dependent proteasomal degradation. OAZ binds ODC monomers and targets them to the proteasome. Here, we report that polyamines, aside from their role in the control of OAZ synthesis and stability, directly enhance OAZ-mediated ODC degradation by the proteasome. Using a stable mutant of OAZ, we show that polyamines promote ODC degradation in Saccharomyces cerevisiae cells even when OAZ levels are not changed. Furthermore, polyamines stimulated the in vitro degradation of ODC by the proteasome in a reconstituted system using purified components. In these assays, spermine shows a greater effect than spermidine. By contrast, polyamines do not have any stimulatory effect on the degradation of ubiquitin-dependent substrates. PMID:28357293

  11. Characterization of an avian histidine decarboxylase and localization of histaminergic neurons in the chicken brain.

    PubMed

    Bessho, Yuki; Iwakoshi-Ukena, Eiko; Tachibana, Tetsuya; Maejima, Sho; Taniuchi, Shusuke; Masuda, Keiko; Shikano, Kenshiro; Kondo, Kunihiro; Furumitsu, Megumi; Ukena, Kazuyoshi

    2014-08-22

    In mammals, it is established that histamine is a neurotransmitter and/or neuromodulator in the central nervous system. It is produced by the enzyme histidine decarboxylase (HDC) in the tuberomammillary nucleus of the posterior hypothalamus. However, HDC as well as histaminergic neurons have not yet been characterized in the avian brain. We have cloned the cDNA for HDC from the chicken hypothalamus and demonstrated that the chicken HDC sequence is highly homologous to the mammalian counterpart, and that the expressed protein shows high enzymatic activity. The expression of HDC mRNA at various sites in the brain was investigated using quantitative RT-PCR. The results showed that the HDC mRNA was highly expressed in the hypothalamic infundibulum. In situ hybridization analyses revealed that the cells containing HDC mRNA were localized in the medial mammillary nucleus of the hypothalamic infundibulum. Intracerebroventricular injection of histamine in chicks resulted in inhibition of feeding behavior. This is the first report of the characterization of histaminergic neurons in the avian brain, and our findings indicate that neuronal histamine exerts anorexigenic effects in chicks.

  12. Tryptamine-induced resistance in tryptophan decarboxylase transgenic poplar and tobacco plants against their specific herbivores.

    PubMed

    Gill, Rishi I S; Ellis, Brian E; Isman, Murray B

    2003-04-01

    The presence of amines and their derivatives in plant tissues is known to influence insect feeding and reproduction. The enzyme tryptophan decarboxylase (TDC) catalyzes the decarboxylation of tryptophan to tryptamine, which is both a bioactive amine and a precursor of other indole derivatives. Transgenic poplar and tobacco plants ectopically expressing TDC1 accumulated elevated levels of tryptamine without affecting plant growth and development. This accumulation was consistently associated with adverse effects on feeding behavior and physiology of Malacosoma disstria Hub. (forest tent caterpillar, FTC) and Manduca sexta L. (tobacco hornworm, THW). Behavior studies with FTC and THW larvae showed that acceptability of the leaf tissue to larvae was inversely related to foliar tryptamine levels. Physiological studies with FTC and THW larvae showed that consumption of leaf tissue from the transgenic lines is deleterious to larvae growth, apparently due to a postingestive mechanism. Thus, ectopic expression of TDC1 can allow sufficient tryptamine to accumulate in poplar and tobacco leaf tissue to suppress significantly the growth of insect pests that normally feed on these plants.

  13. Catecholamines are required for testosterone induction of ornithine decarboxylase in the mouse kidney.

    PubMed

    Cremades, A; Tovar, A; Peñafiel, R

    1992-08-01

    This report presents a study on the influence of the adrenergic system on androgen-regulated ornithine decarboxylase activity (ODC) in the mouse kidney. We have found that the existing levels of renal ODC in male mice and the increase induced by testosterone in male, female, castrated male and hypophysectomized mice were dramatically impaired by catecholamine depletion produced by treatment with either alpha-methyl-p-tyrosine or reserpine. Blockade of beta-receptors by treatment of the animals with the antagonists propranolol or bisoprolol was unable to prevent the effect of testosterone on the renal enzyme. However, alpha i-blockade obtained by treatment with prazosin or phenoxybenzamine was partially effective to produce the decrease of basal ODC in control male or to diminish ODC induction produced by androgens. All these findings demonstrate that catecholamines and alpha 1-adrenergic receptors are implicated in androgen-regulated ODC activity in the mouse kidney and suggest a plausible role of sympathetic renal innervation in enzyme induction by steroid hormones in this organ.

  14. Differential roles of pyruvate decarboxylase in aerial and embedded mycelia of the ascomycete Gibberella zeae.

    PubMed

    Son, Hokyoung; Min, Kyunghun; Lee, Jungkwan; Choi, Gyung Ja; Kim, Jin-Cheol; Lee, Yin-Won

    2012-04-01

    The pyruvate-acetaldehyde-acetate (PAA) pathway has diverse roles in eukaryotes. Our previous study on acetyl-coenzyme A synthetase 1 (ACS1) in Gibberella zeae suggested that the PAA pathway is important for lipid production, which is required for perithecia maturation. In this study, we deleted all three pyruvate decarboxylase (PDC) genes, which encode enzymes that function upstream of ACS1 in the PAA pathway. Results suggest PDC1 is required for lipid accumulation in the aerial mycelia, and deletion of PDC1 resulted in highly wettable mycelia. However, the total amount of lipids in the PDC1 deletion mutants was similar to that of the wild-type strain, likely due to compensatory lipid production processes in the embedded mycelia. PDC1 was expressed both in the aerial and embedded mycelia, whereas ACS1 was observed only in the aerial mycelia in a PDC1-dependent manner. PDC1 is also involved in vegetative growth of embedded mycelia in G. zeae, possibly through initiating the ethanol fermentation pathway. Thus, PDC1 may function as a key metabolic enzyme crucial for lipid production in the aerial mycelia, but play a different role in the embedded mycelia, where it might be involved in energy generation by ethanol fermentation.

  15. Crystal Structure and Pyridoxal 5-Phosphate Binding Property of Lysine Decarboxylase from Selenomonas ruminantium

    PubMed Central

    Sagong, Hye-Young; Son, Hyeoncheol Francis; Kim, Sunghwan; Kim, Yong-Hwan; Kim, Il-Kwon; Kim, Kyung-Jin

    2016-01-01

    Lysine decarboxylase (LDC) is a crucial enzyme for acid stress resistance and is also utilized for the biosynthesis of cadaverine, a promising building block for bio-based polyamides. We determined the crystal structure of LDC from Selenomonas ruminantium (SrLDC). SrLDC functions as a dimer and each monomer consists of two distinct domains; a PLP-binding barrel domain and a sheet domain. We also determined the structure of SrLDC in complex with PLP and cadaverine and elucidated the binding mode of cofactor and substrate. Interestingly, compared with the apo-form of SrLDC, the SrLDC in complex with PLP and cadaverine showed a remarkable structural change at the PLP binding site. The PLP binding site of SrLDC contains the highly flexible loops with high b-factors and showed an open-closed conformational change upon the binding of PLP. In fact, SrLDC showed no LDC activity without PLP supplement, and we suggest that highly flexible PLP binding site results in low PLP affinity of SrLDC. In addition, other structurally homologous enzymes also contain the flexible PLP binding site, which indicates that high flexibility at the PLP binding site and low PLP affinity seems to be a common feature of these enzyme family. PMID:27861532

  16. Cloning and primary structure of a human islet isoform of glutamic acid decarboxylase from chromosome 10

    SciTech Connect

    Karlsen, A.E.; Hagopian, W.A.; Grubin, C.E.; Dube, S.; Disteche, C.M.; Adler, D.A.; Baermeier, H.; Lernmark, A. ); Mathewes, S.; Grant, F.J.; Foster, D. )

    1991-10-01

    Glutamic acid decarboxylase which catalyzes formation of {gamma}-aminobutyric acid from L-glutamic acid, is detectable in different isoforms with distinct electrophoretic and kinetic characteristics. GAD has also been implicated as an autoantigen in the vastly differing autoimmune disease stiff-man syndrome and insulin-dependent diabetes mellitus. Despite the differing GAD isoforms, only one type of GAD cDNA (GAD-1), localized to a syntenic region of chromosome 2, has been isolated from rat, mouse, and cat. Using sequence information from GAD-1 to screen a human pancreatic islet cDNA library, the authors describe the isolation of an additional GAD cDNA (GAD-2), which was mapped to the short arm of human chromosome 10. Genomic Southern blotting with GAD-2 demonstrated a hybridization pattern different form that detected by GAD-1. GAD-2 recognizes a 5.6-kilobase transcript in both islets and brain, in contrast to GAD-1, which detects a 3.7-kilobase transcript in brain only. The deduced 585-amino acid sequence coded for by GAD-2 shows < 65% identify to previously published, highly conserved GAD-1 brain sequences, which show > 96% deduced amino acid sequence homology among the three species.

  17. Overexpression of Actinidia deliciosa pyruvate decarboxylase 1 gene enhances waterlogging stress in transgenic Arabidopsis thaliana.

    PubMed

    Zhang, Ji-Yu; Huang, Sheng-Nan; Wang, Gang; Xuan, Ji-Ping; Guo, Zhong-Ren

    2016-09-01

    Ethanolic fermentation is classically associated with waterlogging tolerance when plant cells switch from respiration to anaerobic fermentation. Pyruvate decarboxylase (PDC), which catalyzes the first step in this pathway, is thought to be the main regulatory enzyme. Here, we cloned a full-length PDC cDNA sequence from kiwifruit, named AdPDC1. We determined the expression of the AdPDC1 gene in kiwifruit under different environmental stresses using qRT-PCR, and the results showed that the increase of AdPDC1 expression during waterlogging stress was much higher than that during salt, cold, heat and drought stresses. Overexpression of kiwifruit AdPDC1 in transgenic Arabidopsis enhanced the resistance to waterlogging stress but could not enhance resistance to cold stress at five weeks old seedlings. Overexpression of kiwifruit AdPDC1 in transgenic Arabidopsis could not enhance resistance to NaCl and mannitol stresses at the stage of seed germination and in early seedlings. These results suggested that the kiwifruit AdPDC1 gene is required during waterlogging but might not be required during other environmental stresses. Expression of the AdPDC1 gene was down-regulated by abscisic acid (ABA) in kiwifruit, and overexpression of the AdPDC1 gene in Arabidopsis inhibited seed germination and root length under ABA treatment, indicating that ABA might negatively regulate the AdPDC1 gene under waterlogging stress.

  18. Cloning and sequence analysis of ornithine decarboxylase gene fragments from the Ascomycota.

    PubMed

    Jiménez-Bremont, Juan Francisco; Rodríguez-Kessler, Margarita; Rodríguez-Guerra, Raul; Cortes-Penagos, Carlos; Torres-Guzman, Juan Carlos; Williamson, June Simpson

    2006-06-01

    Ornithine decarboxylase (ODC; EC 4.1.1.17) catalyzes the initial step in the biosynthesis of polyamines, the conversion of ornithine to putrescine. Based on the most conserved regions of fungal ODCs, we designed and synthesized oligonucleotides to amplify homologous fragments of three important plant pathogenic Pyrenomycete fungi (Ascomycota), Magnaporthe grisea, Colletotrichum lindemuthianum and Fusarium solani, and one insect pathogenic fungus Metarhizium anisopliae. Cloning and sequencing of the amplified fragments revealed homologies of between 37 to 88% with other fungal ODCs. The predicted peptide sequences were compared by Clustal analysis and conserved sequences corresponding to the substrate and cofactor binding sites were identified. Comparative analyses of the ODC fragments isolated in this study, revealed high homology between them (68.3-81.1%) and also with other Pyrenomycetes such as Neurospora crassa (order Sordariales; 68.6-72.9%) and Fusarium graminearum (order Hypocreales; 70.8-88.1%). Data obtained in this work revealed that these fungi constitute a compact group separated from other eukaryotic ODCs.

  19. Immobilization and characterization of benzoylformate decarboxylase from Pseudomonas putida on spherical silica carrier.

    PubMed

    Peper, Stephanie; Kara, Selin; Long, Wei Sing; Liese, Andreas; Niemeyer, Bernd

    2011-08-01

    If an adequate biocatalyst is identified for a specific reaction, immobilization is one possibility to further improve its properties. The immobilization allows easy recycling, improves the enzyme performance, and it often enhances the stability of the enzyme. In this work, the immobilization of the benzoylformate decarboxylase (BFD) variant, BFD A460I-F464I, from Pseudomonas putida was accomplished on spherical silica. Silicagel is characterized by its high mechanical stability, which allows its application in different reactor types without restrictions. The covalently bound enzyme was characterized in terms of its activity, stability, and kinetics for the formation of chiral 2-hydroxypropiophenone (2-HPP) from benzaldehyde and acetaldehyde. Moreover, temperature as well as pressure dependency of immobilized BFD A460I-F464I activity and enantioselectivity were analyzed. The used wide-pore silicagel shows a good accessibility of the immobilized enzyme. The activity of the immobilized BFD A460I-F464I variant was determined to be 70% related to the activity of the free enzyme. Thereby, the enantioselectivity of the enzyme was not influenced by the immobilization. In addition, a pressure-induced change in stereoselectivity was found both for the free and for the immobilized enzyme. With increasing pressure, the enantiomeric excess (ee) of (R)-2-HPP can be increased from 44% (0.1 MPa) to 76% (200 MPa) for the free enzyme and from 43% (0.1 MPa) to 66% (200 MPa) for the immobilized enzyme.

  20. Structural analysis of Bacillus pumilus phenolic acid decarboxylase, a lipocalin-fold enzyme

    SciTech Connect

    Matte, Allan; Grosse, Stephan; Bergeron, Hélène; Abokitse, Kofi; Lau, Peter C.K.

    2012-04-30

    The decarboxylation of phenolic acids, including ferulic and p-coumaric acids, to their corresponding vinyl derivatives is of importance in the flavoring and polymer industries. Here, the crystal structure of phenolic acid decarboxylase (PAD) from Bacillus pumilus strain UI-670 is reported. The enzyme is a 161-residue polypeptide that forms dimers both in the crystal and in solution. The structure of PAD as determined by X-ray crystallography revealed a -barrel structure and two -helices, with a cleft formed at one edge of the barrel. The PAD structure resembles those of the lipocalin-fold proteins, which often bind hydrophobic ligands. Superposition of structurally related proteins bound to their cognate ligands shows that they and PAD bind their ligands in a conserved location within the -barrel. Analysis of the residue-conservation pattern for PAD-related sequences mapped onto the PAD structure reveals that the conservation mainly includes residues found within the hydrophobic core of the protein, defining a common lipocalin-like fold for this enzyme family. A narrow cleft containing several conserved amino acids was observed as a structural feature and a potential ligand-binding site.

  1. A heterozygous deletion in the glutamate decarboxylase 67 gene enhances maternal and fetal stress vulnerability.

    PubMed

    Uchida, Taku; Oki, Yutaka; Yanagawa, Yuchio; Fukuda, Atsuo

    2011-04-01

    Both down-regulation of glutamate decarboxylase 67 (GAD67) and maternal exposure to severe stress during pregnancy can increase the risk of schizophrenia and related psychotic disorders in the offspring. To investigate a gene-environment interaction, we performed the restraint-and-light stress to pregnant GAD67-GFP knock-in (GAD67(+/GFP)) and wild-type (GAD67(+/+)) mice three times a day for 45 min per session during gestational day (G) 15.0-17.5. The stress hormone (corticosterone) level of pregnant GAD67(+/GFP) mice (the overall GABA content is reduced because of the destruction of one allele of the endogenous GAD67 gene) was higher than that of GAD67(+/+), even without stress. The fetal body weights (GAD67(+/+)) in the GAD67(+/GFP) mothers were lower than those in the GAD67(+/+) mothers. GAD67(+/GFP) fetuses exhibited higher corticosterone (CORT) levels than GAD67(+/+) fetuses, even in non-stressed GAD67(+/+) mothers. Fetal body weight-decreases and CORT-increases by maternal stress (GAD67(+/+) mother) were significantly more in the GAD67(+/GFP) fetuses than the GAD67(+/+) fetuses. These results indicate that a GAD67 heterozygous deletion itself enhances vulnerability by many aspects, e.g., maternal stress, maternity, and being in utero. Thus, an abnormality in GAD67 could interact with environmental risk factors of psychiatric disorders, including schizophrenia.

  2. Pathogenic Roles of Glutamic Acid Decarboxylase 65 Autoantibodies in Cerebellar Ataxias

    PubMed Central

    Hampe, Christiane S.

    2017-01-01

    Reports suggesting a pathogenic role of autoantibodies directed against glutamic acid decarboxylase 65 (GAD65Abs) in cerebellar ataxias (CAs) are reviewed, and debatable issues such as internalization of antibodies by neurons and roles of epitopes are discussed. GAD65 is one of two enzymes that catalyze the conversion of glutamate to the inhibitory neurotransmitter gamma-aminobutyric acid (GABA). A pathogenic role of GAD65Ab in CAs is suggested by in vivo and in vitro studies. (1) Intracerebellar administration of cerebrospinal fluid (CSF) immunoglobulins (IgGs) obtained from GAD65Ab-positive CA patients impairs cerebellar modulation of motor control in rats. (2) CSF IgGs act on terminals of GABAergic neurons and decrease the release of GABA in cerebellar slices from rats and mice. (3) Absorption of GAD65Ab by recombinant GAD65 diminishes the above effects, and monoclonal human GAD65Ab (b78) mimic the effects of CSF IgGs in vivo and in vitro. Studies using GAD65-KO mice confirm that the target molecule is GAD65. (4) Notably, the effects of GAD65Ab depend on the epitope specificity of the monoclonal GAD65Ab. Taken together, these results indicate that epitope-specific GAD65Ab-induced impairment of GABA release is involved in the pathogenesis of GAD65Ab-positive CA and support the early detection of GAD65Ab-associated CA to initiate immunotherapy before irreversible neuronal death in the cerebellum. PMID:28386570

  3. Characterization of the p-coumaric acid decarboxylase from Lactobacillus plantarum CECT 748(T).

    PubMed

    Rodríguez, Héctor; Landete, José María; Curiel, José Antonio; de Las Rivas, Blanca; Mancheño, José Miguel; Muñoz, Rosario

    2008-05-14

    It was previously reported that cell cultures from Lactobacillus plantarum CECT 748 (T) were able to decarboxylate phenolic acids, such as p-coumaric, m-coumaric, caffeic, ferulic, gallic, and protocatechuic acid. The p-coumaric acid decarboxylase (PDC) from this strain has been overexpressed and purified. This PDC differs at its C-terminal end when compared to the previously reported PDC from L. plantarum LPCHL2. Because the C-terminal region of PDC is involved in enzymatic activity, especially in substrate activity, it was decided to biochemically characterize the PDC from L. plantarum CECT 748 (T). Contrarily to L. plantarum LPCHL2 PDC, the recombinant PDC from L. plantarum CECT 748 (T) is a heat-labile enzyme, showing optimal activity at 22 degrees C. This PDC is able to decarboxylate exclusively the hydroxycinnamic acids p-coumaric, caffeic, and ferulic acids. Kinetic analysis showed that the enzyme has a 14-fold higher K(M) value for p-coumaric and caffeic acids than for ferulic acid. PDC catalyzes the formation of the corresponding 4-vinyl derivatives (vinylphenol and vinylguaiacol) from p-coumaric and ferulic acids, respectively, which are valuable food additives that have been approved as flavoring agents. The biochemical characteristics showed by L. plantarum PDC should be taken into account for its potential use in the food-processing industry.

  4. Improving single slope ADC and an example implemented in FPGA with 16.7 GHz equivalent counter clook frequency

    SciTech Connect

    Wu, Jinyuan; Odeghe, John; Stackley, Scott; Zha, Charles; /Rice U.

    2011-11-01

    Single slope ADC is a common building block in many ASCI or FPGA based front-end systems due to its simplicity, small silicon footprint, low noise interference and low power consumption. In single slope ADC, using a Gray code counter is a popular scheme for time digitization, in which the comparator output drives the clock (CK) port of a register to latch the bits from the Gray code counter. Unfortunately, feeding the comparator output into the CK-port causes unnecessary complexities and artificial challenges. In this case, the propagation delays of all bits from the counter to the register inputs must be matched and the counter must be a Gray code one. A simple improvement on the circuit topology, i.e., feeding the comparator output into the D-port of a register, will avoid these unnecessary challenges, eliminating the requirement of the propagation delay match of the counter bits and allowing the use of regular binary counters. This scheme not only simplifies current designs for low speeds and resolutions, but also opens possibilities for applications requiring higher speeds and resolutions. A multi-channel single slope ADC based on a low-cost FPGA device has been implemented and tested. The timing measurement bin width in this work is 60 ps, which would need a 16.7 GHz counter clock had it implemented with the conventional Gray code counter scheme. A 12-bit performance is achieved using a fully differential circuit making comparison between the input and the ramping reference, both in differential format.

  5. Could ADC values be a promising diagnostic criterion for differentiating malignant and benign hepatic lesions in Asian populations

    PubMed Central

    Peng, Jing; Li, Jing-Jin; Li, Jiang; Li, He-Wen; Xu, Guo-Ping; Jia, Rong-Rong; Zhang, Xue-Ning; Zhao, Yang

    2016-01-01

    Abstract Background: Liver cancer exhibits geographic and ethnic differences in its prevalence and biology, which implies that it is impractical to develop universal guidelines for all patients. Thus, a meta-analysis was conducted to identify the accuracy of apparent diffusion coefficients (ADCs) for discriminating malignant from benign liver lesions in Asians. Methods: Eligible studies published in PubMed, Ovid, and Embase/Medline were updated onto October 2014. STATA 12.0 and Meta-Disc 1.4 were used to perform this meta-analysis. Results: Eight studies comprising 661 benign liver lesions and 598 malignant liver lesions fulfilled all the inclusion criteria. The pooled sensitivity, specificity, positive likelihood ratio, negative likelihood ratio, and diagnostic odds ratio were 0.88 (95% confidence interval [CI] 0.75–0.95), 0.93 (95% CI 0.86–0.97), 12.42 (95% CI 6.09–25.31), 0.13 (95% CI 0.06–0.29), and 95.58 (95% CI 35.29–258.89), respectively. Overall, the area under the summary receiver-operating characteristic curve was 0.96 (95% CI 0.94–0.98). Heterogeneity was found to originate potentially from the type of benign lesion. A subgroup analysis showed that differentiating between hemangiomas, cysts, and malignant liver lesions produced a significantly higher diagnostic accuracy than that of solid liver lesions. Conclusion: Our meta-analysis indicated that ADC could be promising for characterizing liver lesions among Asians, indicating that the ADC value is a promising diagnostic criterion candidate. Meanwhile, the use of dual b values could be sufficient for liver lesion characterization. However, large-scale, high-quality trials should be conducted to identify specific standards, including cut-off values for further development of diffusion-weighted imaging as a routine clinical application among Asian populations. PMID:27902599

  6. ADCT-301, a Pyrrolobenzodiazepine (PBD) Dimer-Containing Antibody-Drug Conjugate (ADC) Targeting CD25-Expressing Hematological Malignancies.

    PubMed

    Flynn, Michael J; Zammarchi, Francesca; Tyrer, Peter C; Akarca, Ayse U; Janghra, Narinder; Britten, Charles E; Havenith, Carin E G; Levy, Jean-Noel; Tiberghien, Arnaud; Masterson, Luke A; Barry, Conor; D'Hooge, Francois; Marafioti, Teresa; Parren, Paul W H I; Williams, David G; Howard, Philip W; van Berkel, Patrick H; Hartley, John A

    2016-11-01

    Despite the many advances in the treatment of hematologic malignancies over the past decade, outcomes in refractory lymphomas remain poor. One potential strategy in this patient population is the specific targeting of IL2R-α (CD25), which is overexpressed on many lymphoma and leukemic cells, using antibody-drug conjugates (ADC). ADCT-301 is an ADC composed of human IgG1 HuMax-TAC against CD25, stochastically conjugated through a dipeptide cleavable linker to a pyrrolobenzodiazepine (PBD) dimer warhead with a drug-antibody ratio (DAR) of 2.3. ADCT-301 binds human CD25 with picomolar affinity. ADCT-301 has highly potent and selective cytotoxicity against a panel of CD25-expressing human lymphoma cell lines. Once internalized, the released warhead binds in the DNA minor groove and exerts its potent cytotoxic action via the formation of DNA interstrand cross-links. A strong correlation between loss of viability and DNA cross-link formation is demonstrated. DNA damage persists, resulting in phosphorylation of histone H2AX, cell-cycle arrest in G2-M, and apoptosis. Bystander killing of CD25-negative cells by ADCT-301 is also observed. In vivo, a single dose of ADCT-301 results in dose-dependent and targeted antitumor activity against both subcutaneous and disseminated CD25-positive lymphoma models. In xenografts of Karpas 299, which expressed both CD25 and CD30, marked superiority over brentuximab vedotin (Adcetris) is observed. Dose-dependent increases in DNA cross-linking, γ-H2AX, and PBD payload staining were observed in tumors in vivo indicating a role as relevant pharmacodynamic assays. Together, these data support the clinical testing of this novel ADC in patients with CD25-expressing tumors. Mol Cancer Ther; 15(11); 2709-21. ©2016 AACR.

  7. A reference voltage in capacitor-resister hybrid SAR ADC for front-end readout system of CZT detector

    NASA Astrophysics Data System (ADS)

    Wei, Liu; Tingcun, Wei; Bo, Li; Lifeng, Yang; Yongcai, Hu

    2016-01-01

    An on-chip reference voltage has been designed in capacitor-resister hybrid SAR ADC for CZT detector with the TSMC 0.35 μm 2P4M CMOS process. The voltage reference has a dynamic load since using variable capacitors and resistances, which need a large driving ability to deal with the current related to the time and sampling rate. Most of the previous articles about the reference for ADC present only the bandgap part for a low temperature coefficient and high PSRR. However, it is not enough and overall, it needs to consider the output driving ability. The proposed voltage reference is realized by the band-gap reference, voltage generator and output buffer. Apart from a low temperature coefficient and high PSRR, it has the features of a large driving ability and low power consumption. What is more, for CZT detectors application in space, a radiation-hardened design has been considered. The measurement results show that the output reference voltage of the buffer is 4.096 V. When the temperature varied from 0 to 80 °C, the temperature coefficient is 12.2 ppm/°C. The PSRR was -70 dB @ 100 kHz. The drive current of the reference can reach up to 10 mA. The area of the voltage reference in the SAR ADC chip is only 449 × 614 μm2. The total power consumption is only 1.092 mW. Project supported by the National Key Scientific Instrument and Equipment Development Project (No. 2011YQ040082), the National Natural Science Foundation of China (No. 61376034), and the Shaanxi Province Science and Technology Innovation Project (No. 2015KTZDGY03-03).

  8. Functional Characterization of a Novel Member of the Amidohydrolase 2 Protein Family, 2-Hydroxy-1-Naphthoic Acid Nonoxidative Decarboxylase from Burkholderia sp. Strain BC1

    PubMed Central

    Pal Chowdhury, Piyali; Basu, Soumik; Dutta, Arindam

    2016-01-01

    ABSTRACT The gene encoding a nonoxidative decarboxylase capable of catalyzing the transformation of 2-hydroxy-1-naphthoic acid (2H1NA) to 2-naphthol was identified, recombinantly expressed, and purified to homogeneity. The putative gene sequence of the decarboxylase (hndA) encodes a 316-amino-acid protein (HndA) with a predicted molecular mass of 34 kDa. HndA exhibited high identity with uncharacterized amidohydrolase 2 proteins of various Burkholderia species, whereas it showed a modest 27% identity with γ-resorcylate decarboxylase, a well-characterized nonoxidative decarboxylase belonging to the amidohydrolase superfamily. Biochemically characterized HndA demonstrated strict substrate specificity toward 2H1NA, whereas inhibition studies with HndA indicated the presence of zinc as the transition metal center, as confirmed by atomic absorption spectroscopy. A three-dimensional structural model of HndA, followed by docking analysis, identified the conserved metal-coordinating and substrate-binding residues, while their importance in catalysis was validated by site-directed mutagenesis. IMPORTANCE Microbial nonoxidative decarboxylases play a crucial role in the metabolism of a large array of carboxy aromatic chemicals released into the environment from a variety of natural and anthropogenic sources. Among these, hydroxynaphthoic acids are usually encountered as pathway intermediates in the bacterial degradation of polycyclic aromatic hydrocarbons. The present study reveals biochemical and molecular characterization of a 2-hydroxy-1-naphthoic acid nonoxidative decarboxylase involved in an alternative metabolic pathway which can be classified as a member of the small repertoire of nonoxidative decarboxylases belonging to the amidohydrolase 2 family of proteins. The strict substrate specificity and sequence uniqueness make it a novel member of the metallo-dependent hydrolase superfamily. PMID:27068590

  9. A 12 bit 40 MSPS SAR ADC with a redundancy algorithm and digital calibration for the ATLAS LAr calorimeter readout

    NASA Astrophysics Data System (ADS)

    Zeloufi, M.

    2016-01-01

    We present a SAR ADC with a generalized redundant search algorithm offering the flexibility to relax the requirements on the DAC settling time. The redundancy also allows a digital background calibration, based on a code density analysis, to compensate for the capacitor mismatch effects. The total number of capacitors used in this architecture is limited to one half of the one in a classical SAR design. Only 211 unit capacitors were necessary to reach 12 bit resolution, and the switching algorithm is intrinsically monotonic. The design is fully differential featuring 12 bit 40 MS/s in a CMOS 130 nm 1P8M process.

  10. A 0.23 pJ 11.05-bit ENOB 125-MS/s pipelined ADC in a 0.18 μm CMOS process

    NASA Astrophysics Data System (ADS)

    Yong, Wang; Jianyun, Zhang; Rui, Yin; Yuhang, Zhao; Wei, Zhang

    2015-05-01

    This paper describes a 12-bit 125-MS/s pipelined analog-to-digital converter (ADC) that is implemented in a 0.18 μm CMOS process. A gate-bootstrapping switch is used as the bottom-sampling switch in the first stage to enhance the sampling linearity. The measured differential and integral nonlinearities of the prototype are less than 0.79 least significant bit (LSB) and 0.86 LSB, respectively, at the full sampling rate. The ADC exhibits an effective number of bits (ENOB) of more than 11.05 bits at the input frequency of 10.5 MHz. The ADC also achieves a 10.5 bits ENOB with the Nyquist input frequency at the full sample rate. In addition, the ADC consumes 62 mW from a 1.9 V power supply and occupies 1.17 mm2, which includes an on-chip reference buffer. The figure-of-merit of this ADC is 0.23 pJ/step. Project supported by the Foundation of Shanghai Municipal Commission of Economy and Informatization (No. 130311).

  11. A 12-bit compact column-parallel SAR ADC with dynamic power control technique for high-speed CMOS image sensors

    NASA Astrophysics Data System (ADS)

    Quanliang, Li; Liyuan, Liu; Ye, Han; Zhongxiang, Cao; Nanjian, Wu

    2014-10-01

    This paper presents a 12-bit column-parallel successive approximation register analog-to-digital converter (SAR ADC) for high-speed CMOS image sensors. A segmented binary-weighted switched capacitor digital-to-analog converter (CDAC) and a staggered structure MOM unit capacitor is used to reduce the ADC area and to make its layout fit double pixel pitches. An electrical field shielding layout method is proposed to eliminate the parasitic capacitance on the top plate of the unit capacitor. A dynamic power control technique is proposed to reduce the power consumption of a single channel during readout. An off-chip foreground digital calibration is adopted to compensate for the nonlinearity due to the mismatch of unit capacitors among the CDAC. The prototype SAR ADC is fabricated in a 0.18 μm 1P5M CIS process. A single SAR ADC occupies 20 × 2020 μm2. Sampling at 833 kS/s, the measured differential nonlinearity, integral nonlinearity and effective number of bits of SAR ADC with calibration are 0.9/-1 LSB, 1/-1.1 LSB and 11.24 bits, respectively; the power consumption is only 0.26 mW under a 1.8-V supply and decreases linearly as the frame rate decreases.

  12. Treatment of idiopathic parkinsonism with L-dopa in the absence and presence of decarboxylase inhibitors: effects on plasma levels of L-dopa, dopa decarboxylase, catecholamines and 3-O-methyl-dopa.

    PubMed

    Boomsma, F; Meerwaldt, J D; Man in't Veld, A J; Hovestadt, A; Schalekamp, M A

    1989-05-01

    The effect of levodopa (L-dopa), alone or in combination with a peripheral decarboxylase inhibitor (PDI), on plasma levels of aromatic-L-amino acid decarboxylase (ALAAD, = dopa decarboxylase), L-dopa, 3-O-methyl-dopa (3-OMD), dopamine (DA), noradrenaline, adrenaline and dopamine beta-hydroxylase has been studied. In healthy subjects and in patients with parkinsonism plasma ALAAD level fell after administration of L-dopa + benserazide, but returned to previous levels within 90 min. In a cross-sectional study blood was obtained, 2 h after dosing, from 104 patients with idiopathic parkinsonism, divided into four groups: no L-dopa treatment (group 1), L-dopa alone (group 2), L-dopa + benserazide (Madopar) (group 3) and L-dopa + carbidopa (Sinemet) (group 4). Plasma ALAAD, which was normal in groups 1 and 2, was increased 3-fold in groups 3 and 4, indicating that there was induction of ALAAD by the co-administration of PDI. Despite this induction of ALAAD, in groups 3 and 4, with half the daily L-dopa dose compared with group 2, plasma L-dopa and 3-OMD levels were 5 times higher, while plasma DA levels were not different. The DA/L-dopa ratio was decreased 5-fold in group 2 and 16-fold in groups 3 and 4 as compared with group 1. Neither 3-OMD levels nor 3-OMD/L-dopa ratios correlated with the occurrence of on-off fluctuations. In a longitudinal study of three patients started on Madopar treatment the induction of plasma ALAAD was found to occur gradually over 3-4 weeks. Further detailed pharmacokinetic studies in plasma and cerebrospinal fluid are required in order to elucidate whether the ALAAD induction by PDI may be related to the loss of clinical efficacy of combination therapy in some patients and how it is related to end-of-dose deterioration and on-off phenomena.

  13. SEMICONDUCTOR INTEGRATED CIRCUITS: An 8-b 300MS/S folding and interpolating ADC for embedded applications

    NASA Astrophysics Data System (ADS)

    Yan, Lu; Li, Lin; Jiefeng, Xia; Fan, Ye; Junyan, Ren

    2010-06-01

    A 1.4-V 8-bit 300-MS/s folding and interpolating analog-to-digital converter (ADC) is proposed. Fabricated in the 0.13-μm CMOS process and occupying only 0.6-mm2 active area, the ADC is especially suitable for embedded applications. The system is optimized for a low-power purpose. Pipelining sampling switches help to cut down the extra power needed for complete settling. An averaging resistor array is placed between two folding stages for power-saving considerations. The converter achieves 43.4-dB signal-to-noise and distortion ratio and 53.3-dB spurious-free dynamic range at 1-MHz input and 42.1-dB and 49.5-dB for Nyquist input. Measured results show a power dissipation of 34 mW and a figure of merit of 1.14 pJ/convstep at 250-MHz sampling rate at 1.4-V supply.

  14. A 0.9-V switched-opamp-based delta—sigma ADC with dual cycle shift DWA

    NASA Astrophysics Data System (ADS)

    Jinchen, Zhao; Menglian, Zhao; Xiaobo, Wu; Hanqing, Wang

    2013-06-01

    This paper presents a low-power high-precision switched-opamp(SO)-based delta—sigma (ΔΣ) analog-to-digital converter (ADC). The proposed SO design allows circuit operation at sub-1 V supply voltage, only needs to work in half of a clock cycle, and thus is suitable for low power applications. In addition, an opamp-sharing technique is applied to save on hardware overheads. Due to the use of a dual cycle shift data weighted averaging (DCS-DWA) technique, mismatch errors caused in the feedback DAC have been eliminated without introducing signal-dependent tones. The proposed ADC has been implemented in a standard 0.18 μm process and measured to have a 92.2 dB peak SNDR and 94.1 dB dynamic range with 25 kHz signal bandwidth. The power consumption is 58 μW for the modulator at 0.9 V supply voltage and 96 μW for the decimation filter, which translate to the figure-of-merit (FOM) of 35.4 fJ/step for the solo modulator, and 94 fJ/step for the whole system.

  15. A 10-bit 80-MS/s opamp-sharing pipelined ADC with a switch-embedded dual-input MDAC

    NASA Astrophysics Data System (ADS)

    Rui, Yin; Youchun, Liao; Wei, Zhang; Zhangwen, Tang

    2011-02-01

    A 10-bit 80-MS/s opamp-sharing pipelined ADC is implemented in a 0.18-μm CMOS. An opamp-sharing MDAC with a switch-embedded dual-input opamp is proposed to eliminate the non-resetting and successive-stage crosstalk problems observed in the conventional opamp-sharing technique. The ADC achieves a peak SNDR of 60.1 dB (ENOB = 9.69 bits) and a peak SFDR of 76 dB, while maintaining more than 9.6 ENOB for the full Nyquist input bandwidth. The core area of the ADC is 1.1 mm2 and the chip consumes 28 mW with a 1.8 V power supply.

  16. A 12-bit 60-MS/s 36-mW SHA-less opamp-sharing pipeline ADC in 130 nm CMOS

    NASA Astrophysics Data System (ADS)

    Wen, X.; Chen, J.; You, Y.; Feng, Y.; Tang, Y.; Zuo, Z.; Vosooghi, B.; Fan, Q.; Xiao, L.; Gong, D.; Liu, T.; Ye, J.

    2016-01-01

    This paper presents a 12-bit 60-MS/s SHA-less opamp-sharing pipeline analog-to-digital converter (ADC) implemented in a 0.13-μ m CMOS technology. A switch-embedded dual-input current-reused operational transconductance amplifier (OTA) with an overlapping two-phase clocking scheme is proposed to achieve low power consumption and eliminate the non-resetting and memory effects observed in conventional opamp-sharing techniques. To further reduce the power consumption, the ADC also incorporates a SHA-less multi-bit structure. The ADC achieves a signal-to-noise and distortion ratio of 64.9 dB and a spurious-free dynamic range of 77.1 dB at 60 MS/s. It occupies 2.3 mm 2 of area and consumes 36 mW of power under a 1.2-V supply.

  17. Mesomere-derived glutamate decarboxylase-expressing blastocoelar mesenchyme cells of sea urchin larvae

    PubMed Central

    Katow, Hideki; Katow, Tomoko; Abe, Kouki; Ooka, Shioh; Kiyomoto, Masato; Hamanaka, Gen

    2014-01-01

    Summary The ontogenetic origin of blastocoelar glutamate decarboxylase (GAD)-expressing cells (GADCs) in larvae of the sea urchin Hemicentrotus pulcherrimus was elucidated. Whole-mount in situ hybridisation (WISH) detected transcription of the gene that encodes GAD in H. pulcherrimus (Hp-gad) in unfertilised eggs and all blastomeres in morulae. However, at and after the swimming blastula stage, the transcript accumulation was particularly prominent in clumps of ectodermal cells throughout the embryonic surface. During the gastrula stage, the transcripts also accumulated in the endomesoderm and certain blastocoelar cells. Consistent with the increasing number of Hp-gad transcribing cells, immunoblot analysis indicated that the relative abundance of Hp-Gad increased considerably from the early gastrula stage until the prism stage. The expression pattern of GADCs determined by immunohistochemistry was identical to the pattern of Hp-gad transcript accumulation determined using WISH. In early gastrulae, GADCs formed blastocoelar cell aggregates around the blastopore with primary mesenchyme cells. The increase in the number of blastocoelar GADCs was inversely proportional to the number of ectodermal GADCs ranging from a few percent of total GADCs in early gastrulae to 80% in late prism larvae; this depended on ingression of ectodermal GADCs into the blastocoel. Some of the blastocoelar GADCs were fluorescein-positive in the larvae that developed from the 16-cell stage chimeric embryos; these comprised fluorescein-labeled mesomeres and unlabelled macromeres and micromeres. Our finding indicates that some of the blastocoelar GADCs are derived from the mesomeres and thus they are the new group of mesenchyme cells, the tertiary mesenchyme cells. PMID:24357228

  18. Sequestered end products and enzyme regulation: the case of ornithine decarboxylase.

    PubMed Central

    Davis, R H; Morris, D R; Coffino, P

    1992-01-01

    The polyamines (putrescine, spermidine, and spermine) are synthesized by almost all organisms and are universally required for normal growth. Ornithine decarboxylase (ODC), an initial enzyme of polyamine synthesis, is one of the most highly regulated enzymes of eucaryotic organisms. Unusual mechanisms have evolved to control ODC, including rapid, polyamine-mediated turnover of the enzyme and control of the synthetic rate of the protein without change of its mRNA level. The high amplitude of regulation and the rapid variation in the level of the protein led biochemists to infer that polyamines had special cellular roles and that cells maintained polyamine concentrations within narrow limits. This view was sustained in part because of our continuing uncertainty about the actual biochemical roles of polyamines. In this article, we challenge the view that ODC regulation is related to precise adjustment of polyamine levels. In no organism does ODC display allosteric feedback inhibition, and in three types of organism, bacteria, fungi, and mammals, the size of polyamine pools may vary radically without having a profound effect on growth. We suggest that the apparent stability of polyamine pools in unstressed cells is due to their being largely bound to cellular polyanions. We further speculate that allosteric feedback inhibition, if it existed, would be inappropriately responsive to changes in the small, freely diffusible polyamine pool. Instead, mechanisms that control the amount of the ODC protein have appeared in most organisms, and even these are triggered inappropriately by variation of the binding of polyamines to ionic binding sites. In fact, feedback inhibition of ODC might be maladaptive during hypoosmotic stress or at the onset of growth, when organisms appear to require rapid increases in the size of their cellular polyamine pools. PMID:1620066

  19. Role of modulation on the effect of microwaves on ornithine decarboxylase activity in L929 cells

    SciTech Connect

    Penafiel, L.M.; Litovitz, T.; Krause, D.; Desta, A.; Mullins, J.M.

    1997-05-01

    The effect of 835 MHz microwaves on the activity of ornithine decarboxylase (ODC) in L929 murine cells was investigated at an SAR of {approximately}2.5 W/kg. The results depended upon the type of modulation employed. AM frequencies of 16 Hz and 60 Hz produced a transient increase in ODC activity that reached a peak at 8 h of exposure and returned to control levels after 24 h of exposure. In this case, ODC was increased by a maximum of 90% relative to control levels. A 40% increase in ODC activity was also observed after 8 h of exposure with a typical signal from a TDMA digital cellular telephone operating in the middle of its transmission frequency range. This signal was burst modulated at 50 Hz, with approximately 30% duty cycle. By contrast, 8 h exposure with 835 MHz microwaves amplitude modulated with speech produced no significant change in ODC activity. Further investigations, with 8 h of exposure to AM microwaves, as a function of modulation frequency, revealed that the response is frequency dependent, decreasing sharply at 6 Hz and 600 Hz. Exposure with 835 MHz microwaves, frequency modulated with a 60 Hz sinusoid, yielded no significant enhancement in ODC activity for exposure times ranging between 2 and 24 h. Similarly, exposure with a typical signal from an AMPS analog cellular telephone, which uses a form of frequency modulation, produced no significant enhancement in ODC activity. Exposure with 835 MHz continuous wave microwaves produced no effects for exposure times between 2 and 24 h, except for a small but statistically significant enhancement in ODC activity after 6 h of exposure.

  20. Substrate Distortion and the Catalytic Reaction Mechanism of 5-Carboxyvanillate Decarboxylase

    PubMed Central

    2015-01-01

    5-Carboxyvanillate decarboxylase (LigW) catalyzes the conversion of 5-carboxyvanillate to vanillate in the biochemical pathway for the degradation of lignin. This enzyme was shown to require Mn2+ for catalytic activity and the kinetic constants for the decarboxylation of 5-carboxyvanillate by the enzymes from Sphingomonas paucimobilis SYK-6 (kcat = 2.2 s–1 and kcat/Km = 4.0 × 104 M–1 s–1) and Novosphingobium aromaticivorans (kcat = 27 s–1 and kcat/Km = 1.1 × 105 M–1 s–1) were determined. The three-dimensional structures of both enzymes were determined in the presence and absence of ligands bound in the active site. The structure of LigW from N. aromaticivorans, bound with the substrate analogue, 5-nitrovanillate (Kd = 5.0 nM), was determined to a resolution of 1.07 Å. The structure of this complex shows a remarkable enzyme-induced distortion of the nitro-substituent out of the plane of the phenyl ring by approximately 23°. A chemical reaction mechanism for the decarboxylation of 5-carboxyvanillate by LigW was proposed on the basis of the high resolution X-ray structures determined in the presence ligands bound in the active site, mutation of active site residues, and the magnitude of the product isotope effect determined in a mixture of H2O and D2O. In the proposed reaction mechanism the enzyme facilitates the transfer of a proton to C5 of the substrate prior to the decarboxylation step. PMID:26714575

  1. Effects of immunization with natural and recombinant lysine decarboxylase on canine gingivitis development.

    PubMed

    Peters, Jennifer L; DeMars, Paul L; Collins, Lindsay M; Stoner, Julie A; Matsumoto, Hiroyuki; Komori, Naoka; Singh, Anil; Feasley, Christa L; Haddock, James A; Levine, Martin

    2012-10-19

    Periodontal disease, gingival inflammation (gingivitis) and periodontal attachment loss (periodontitis), causes tooth loss and susceptibility to chronic inflammation. Professionally scaling and cleaning the teeth regularly controls the disease, but is expensive in companion animals. Eikenella corrodens is common in canine oral cavities where it is a source of lysine decarboxylase (LDC). In human dental biofilms (plaques), LDC converts lysine to cadaverine and impairs the gingival epithelial barrier to bacteria. LDC vaccination may therefore retard gingivitis development. Year-old beagle dogs provided blood samples, and had weight and clinical measurements (biofilm and gingivitis) recorded. After scaling and cleaning, two dogs were immunized subcutaneously with 0.2mg native LDC from E. corrodens and 2 sets of four dogs with 0.2mg recombinant LDC purified from Escherichia coli. A third set of 4 dogs was immunized intranasally. Rehydragel(®), Emulsigen(®), Polygen™ or Carbigen™ were used as adjuvant. Four additional pairs of dogs were sham-immunized with each adjuvant alone (controls). Immunizations were repeated twice, 3 weeks apart, and clinical measurements were obtained after another 2 weeks, when the teeth were scaled and cleaned again. Tooth brushing was then stopped and the diet was changed from hard to soft chow. Clinical measurements were repeated after 1, 2, 3, 4, 6 and 8 weeks. Compared with sham-immunized dogs, gingivitis was reduced over all 8 weeks of soft diet after subcutaneous immunization with native LDC, or after intranasal immunization with recombinant LDC in Carbigen™, but for only 6 of the 8 weeks after subcutaneous immunization with recombinant LDC in Emulsigen(®) (repeated measures ANOVA). Subcutaneous vaccination induced a strong serum IgG antibody response that decreased during the soft diet period, whereas intranasal immunization induced a weak serum IgA antibody response that did not decrease. Immunization with recombinant LDC may

  2. Glutamate Decarboxylase 1 Overexpression as a Poor Prognostic Factor in Patients with Nasopharyngeal Carcinoma

    PubMed Central

    Lee, Yi-Ying; Chao, Tung-Bo; Sheu, Ming-Jen; Tian, Yu-Feng; Chen, Tzu-Ju; Lee, Sung-Wei; He, Hong-Lin; Chang, I-Wei; Hsing, Chung-Hsi; Lin, Ching-Yih; Li, Chien-Feng

    2016-01-01

    Background: Glutamate decarboxylase 1 (GAD1) which serves as a rate-limiting enzyme involving in the production of γ-aminobutyric acid (GABA), exists in the GABAergic neurons in the central nervous system (CNS). Little is known about the relevance of GAD1 to nasopharyngeal carcinoma (NPC). Through data mining on a data set derived from a published transcriptome database, this study first identified GAD1 as a differentially upregulated gene in NPC. We aimed to evaluate GAD1 expression and its prognostic effect on patients with early and locoregionally advanced NPC. Methods: We evaluated GAD1 immunohistochemistry and performed an H-score analysis on biopsy specimens from 124 patients with nonmetastasized NPC receiving treatment. GAD1 overexpression was defined as an H score higher than the median value. The findings of such an analysis are correlated with clinicopathological behaviors and survival rates, namely disease-specific survival (DSS), distant-metastasis-free survival (DMeFS), and local recurrence-free survival (LRFS) rates. Results: GAD1 overexpression was significantly associated with an increase in the primary tumor status (p < 0.001) and American Joint Committee on Cancer (AJCC) stages III-IV (p = 0.002) and was a univariate predictor of adverse outcomes of DSS (p = 0.002), DMeFS (p < 0.0001), and LRFS (p = 0.001). In the multivariate comparison, in addition to advanced AJCC stages III-IV, GAD1 overexpression remained an independent prognosticator of short DSS (p = 0.004, hazard ratio = 2.234), DMeFS (p < 0.001, hazard ratio = 4.218), and LRFS (p = 0.013, hazard ratio = 2.441) rates. Conclusions: Our data reveal that GAD1 overexpression was correlated with advanced disease status and may thus be a critical prognostic indicator of poor outcomes in NPC and a potential therapeutic target to facilitate the development of effective treatment modalities. PMID:27698909

  3. Structural Basis for Nucleotide Binding and Reaction Catalysis in Mevalonate Diphosphate Decarboxylase

    SciTech Connect

    Barta, Michael L.; McWhorter, William J.; Miziorko, Henry M.; Geisbrecht, Brian V.

    2012-09-17

    Mevalonate diphosphate decarboxylase (MDD) catalyzes the final step of the mevalonate pathway, the Mg{sup 2+}-ATP dependent decarboxylation of mevalonate 5-diphosphate (MVAPP), producing isopentenyl diphosphate (IPP). Synthesis of IPP, an isoprenoid precursor molecule that is a critical intermediate in peptidoglycan and polyisoprenoid biosynthesis, is essential in Gram-positive bacteria (e.g., Staphylococcus, Streptococcus, and Enterococcus spp.), and thus the enzymes of the mevalonate pathway are ideal antimicrobial targets. MDD belongs to the GHMP superfamily of metabolite kinases that have been extensively studied for the past 50 years, yet the crystallization of GHMP kinase ternary complexes has proven to be difficult. To further our understanding of the catalytic mechanism of GHMP kinases with the purpose of developing broad spectrum antimicrobial agents that target the substrate and nucleotide binding sites, we report the crystal structures of wild-type and mutant (S192A and D283A) ternary complexes of Staphylococcus epidermidis MDD. Comparison of apo, MVAPP-bound, and ternary complex wild-type MDD provides structural information about the mode of substrate binding and the catalytic mechanism. Structural characterization of ternary complexes of catalytically deficient MDD S192A and D283A (k{sub cat} decreased 10{sup 3}- and 10{sup 5}-fold, respectively) provides insight into MDD function. The carboxylate side chain of invariant Asp{sup 283} functions as a catalytic base and is essential for the proper orientation of the MVAPP C3-hydroxyl group within the active site funnel. Several MDD amino acids within the conserved phosphate binding loop ('P-loop') provide key interactions, stabilizing the nucleotide triphosphoryl moiety. The crystal structures presented here provide a useful foundation for structure-based drug design.

  4. Functional Roles of the Dimer-Interface Residues in Human Ornithine Decarboxylase

    PubMed Central

    Lee, Chien-Yun; Liu, Yi-Liang; Lin, Chih-Li; Liu, Guang-Yaw; Hung, Hui-Chih

    2014-01-01

    Ornithine decarboxylase (ODC) catalyzes the decarboxylation of ornithine to putrescine and is the rate-limiting enzyme in the polyamine biosynthesis pathway. ODC is a dimeric enzyme, and the active sites of this enzyme reside at the dimer interface. Once the enzyme dissociates, the enzyme activity is lost. In this paper, we investigated the roles of amino acid residues at the dimer interface regarding the dimerization, protein stability and/or enzyme activity of ODC. A multiple sequence alignment of ODC and its homologous protein antizyme inhibitor revealed that 5 of 9 residues (residues 165, 277, 331, 332 and 389) are divergent, whereas 4 (134, 169, 294 and 322) are conserved. Analytical ultracentrifugation analysis suggested that some dimer-interface amino acid residues contribute to formation of the dimer of ODC and that this dimerization results from the cooperativity of these interface residues. The quaternary structure of the sextuple mutant Y331S/Y389D/R277S/D332E/V322D/D134A was changed to a monomer rather than a dimer, and the Kd value of the mutant was 52.8 µM, which is over 500-fold greater than that of the wild-type ODC (ODC_WT). In addition, most interface mutants showed low but detectable or negligible enzyme activity. Therefore, the protein stability of these interface mutants was measured by differential scanning calorimetry. These results indicate that these dimer-interface residues are important for dimer formation and, as a consequence, are critical for enzyme catalysis. PMID:25140796

  5. Ornithine decarboxylase activity as a marker of androgen and antiandrogen action in the rat epididymis.

    PubMed

    de las Heras, M A; Suescun, M O; Calandra, R S

    1988-05-01

    After castration, there was a marked decrease in serum androgen concentration at 6 h, and a dramatic inhibition of ornithine decarboxylase (ODC) at 12 h. Administration of testosterone propionate to castrated rats at a dose of 0.05 mg/animal restored ODC activity to the normal value. However, no change was observed when intact rats were treated with testosterone even at a 40-fold higher dose, indicating that endogenous androgens present in intact rats are far in excess for maintenance of maximal levels of activity. Administration of the antiandrogen flutamide to intact rats caused a moderate decrease in epididymal weight, whereas this effect was more pronounced in castrated, androgen-treated rats. In the latter, the effect of flutamide was significant at the lowest dose used (0.5 mg/day). ODC activity was significantly decreased by flutamide treatment of intact rats, but even at the highest dose used (10 mg/day) only a 39% inhibition was observed. In flutamide-treated rats, LH concentrations were markedly increased, as were serum and epididymal androgens. In androgen-treated castrated rats, flutamide caused epididymal ODC to fall to undetectable values. These results show that: (1) androgens are essential for the maintenance of ODC activity in the epididymis; (2) epididymal ODC activity is maximally stimulated by endogenous androgens, at least in the pubertal rat; (3) the apparent potency of flutamide is substantially lowered by an increase in epididymal androgens. We suggest that ODC is a sensitive marker of the action of androgens and antiandrogens in the epididymis.

  6. Snapshot of a Reaction Intermediate: Analysis of Benzoylformate Decarboxylase in Complex with a Benzoylphosphonate Inhibitor

    SciTech Connect

    Brandt, Gabriel S.; Kneen, Malea M.; Chakraborty, Sumit; Baykal, Ahmet T.; Nemeria, Natalia; Yep, Alejandra; Ruby, David I.; Petsko, Gregory A.; Kenyon, George L.; McLeish, Michael J.; Jordan, Frank; Ringe, Dagmar

    2009-04-22

    Benzoylformate decarboxylase (BFDC) is a thiamin diphosphate- (ThDP-) dependent enzyme acting on aromatic substrates. In addition to its metabolic role in the mandelate pathway, BFDC shows broad substrate specificity coupled with tight stereo control in the carbon-carbon bond-forming reverse reaction, making it a useful biocatalyst for the production of chiral-hydroxy ketones. The reaction of methyl benzoylphosphonate (MBP), an analogue of the natural substrate benzoylformate, with BFDC results in the formation of a stable analogue (C2{alpha}-phosphonomandelyl-ThDP) of the covalent ThDP-substrate adduct C2{alpha}-mandelyl-ThDP. Formation of the stable adduct is confirmed both by formation of a circular dichroism band characteristic of the 1',4'-iminopyrimidine tautomeric form of ThDP (commonly observed when ThDP forms tetrahedral complexes with its substrates) and by high-resolution mass spectrometry of the reaction mixture. In addition, the structure of BFDC with the MBP inhibitor was solved by X-ray crystallography to a spatial resolution of 1.37 {angstrom} (PDB ID 3FSJ). The electron density clearly shows formation of a tetrahedral adduct between the C2 atom of ThDP and the carbonyl carbon atom of the MBP. This adduct resembles the intermediate from the penultimate step of the carboligation reaction between benzaldehyde and acetaldehyde. The combination of real-time kinetic information via stopped-flow circular dichroism with steady-state data from equilibrium circular dichroism measurements and X-ray crystallography reveals details of the first step of the reaction catalyzed by BFDC. The MBP-ThDP adduct on BFDC is compared to the recently solved structure of the same adduct on benzaldehyde lyase, another ThDP-dependent enzyme capable of catalyzing aldehyde condensation with high stereospecificity.

  7. Role of modulation on the effect of microwaves on ornithine decarboxylase activity in L929 cells.

    PubMed

    Penafiel, L M; Litovitz, T; Krause, D; Desta, A; Mullins, J M

    1997-01-01

    The effect of 835 MHz microwaves on the activity of ornithine decarboxylase (ODC) in L929 murine cell was investigated at an SAR of approximately 2.5 W/kg. The results depended upon the type of modulation employed. AM frequencies of 16 Hz and 60 Hz produced a transient increase in ODC activity that reached a peak at 8 h of exposure and returned to control levels after 24 h of exposure. In this case, ODC was increased by a maximum of 90% relative to control levels. A 40% increase in ODC activity was also observed after 8 h of exposure with a typical signal from a TDMA digital cellular telephone operating in the middle of its transmission frequency range (approximately 840 MHz). This signal was burst modulated at 50 Hz, with approximately 30% duty cycle. By contrast, 8 h exposure with 835 MHz microwaves amplitude modulated with speech produced no significant change in ODC activity. Further investigations, with 8 h of exposure to AM microwaves, as a function of modulation frequency, revealed that the response is frequency dependent, decreasing sharply at 6 Hz an 600 Hz. Exposure with 835 MHz microwaves, frequency modulated with a 60 Hz sinusoid, yielded no significant enhancement in ODC activity for exposure times ranging between 2 and 24 h. Similarly, exposure with a typical signal from an AMPS analog cellular telephone, which uses a form of frequency modulation, produced no significant enhancement in ODC activity. Exposure with 835 MHz continuous wave microwaves produced no effects for exposure times between 2 and 24 h, except for a small but statistically significant enhancement in ODC activity after 6 h of exposure. Comparison of these results suggests that effects are much more robust when the modulation causes low-frequency periodic changes in the amplitude of the microwave carrier.

  8. Uroporphyrinogen decarboxylase: Complete human gene sequence and molecular study of three families with hepatoerythropoietic porphyria

    SciTech Connect

    Moran-Jimenez, M.J.; Ged, C.; Verneuil, H. de

    1996-04-01

    A deficiency in uroporphyrinogen decarboxylase (UROD) enzyme activity, the fifth enzyme of the heme biosynthetic pathway, is found in patients with sporadic porphyria cutanea tarda (s-PCT), familial porphyria cutanea tarda (f-PCT), and hepatoerythropoietic porphyria (HEP). Subnormal UROD activity is due to mutations of the UROD gene in both f-PCT and HEP, but no mutations have been found in s-PCT. Genetic analysis has determined that f-PCT is transmitted as an autosomal dominant trait. In contrast, HEP, a severe form of cutaneous porphyria, is transmitted as an autosomal recessive trait. HEP is characterized by a profound deficiency of UROD activity, and the disease is usually manifest in childhood. In this study, a strategy was designed to identify alleles responsible for the HEP phenotype in three unrelated families. Mutations of UROD were identified by direct sequencing of four amplified fragments that contained the entire coding sequence of the UROD gene. Two new missense mutations were observed at the homoallelic state: P62L (proline-to-leucine substitution at codon 62) in a Portuguese family and Y311C (tyrosine-to-cysteine substitution at codon 311) in an Italian family. A third mutation, G281E, was observed in a Spanish family. This mutation has been previously described in three families from Spain and one from Tunisia. In the Spanish family described in this report, a paternal uncle of the proband developed clinically overt PCT as an adult and proved to be heterozygous for the G281E mutation. Mutant cDNAs corresponding to the P62L and Y311C changes detected in these families were created by site-directed mutagenesis. Recombinant proteins proved to have subnormal enzyme activity, and the Y311C mutant was thermolabile. 24 refs., 7 figs., 4 tabs.

  9. Epilepsy and hippocampal neurodegeneration induced by glutamate decarboxylase inhibitors in awake rats.

    PubMed

    Salazar, Patricia; Tapia, Ricardo

    2015-10-01

    Glutamic acid decarboxylase (GAD), the enzyme responsible for GABA synthesis, requires pyridoxal phosphate (PLP) as a cofactor. Thiosemicarbazide (TSC) and γ-glutamyl-hydrazone (PLPGH) inhibit the free PLP-dependent isoform (GAD65) activity after systemic administration, leading to epilepsy in mice and in young, but not in adult rats. However, the competitive GAD inhibitor 3-mercaptopropionic acid (MPA) induces convulsions in both immature and adult rats. In the present study we tested comparatively the epileptogenic and neurotoxic effects of PLPGH, TSC and MPA, administered by microdialysis in the hippocampus of adult awake rats. Cortical EEG and motor behavior were analyzed during the next 2h, and aspartate, glutamate and GABA were measured by HPLC in the microdialysis-collected fractions. Twenty-four hours after drug administration rats were fixed for histological analysis of the hippocampus. PLPGH or TSC did not affect the motor behavior, EEG or cellular morphology, although the extracellular concentration of GABA was decreased. In contrast, MPA produced intense wet-dog shakes, EEG epileptiform discharges, a >75% reduction of extracellular GABA levels and remarkable neurodegeneration of the CA1 region, with >80% neuronal loss. The systemic administration of the NMDA glutamate receptor antagonist MK-801 30 min before MPA did not prevent the MPA-induced epilepsy but significantly protected against its neurotoxic effect, reducing neuronal loss to <30%. We conclude that in adult awake rats, drugs acting on PLP availability have only a weak effect on GABA neurotransmission, whereas direct GAD inhibition produced by MPA induces hyperexcitation leading to epilepsy and hippocampal neurodegeneration. Because this degeneration was prevented by the blockade of NMDA receptors, we conclude that it is due to glutamate-mediated excitotoxicity consequent to disinhibition of the hippocampal excitatory circuits.

  10. Developmental PCB Exposure Increases Audiogenic Seizures and Decreases Glutamic Acid Decarboxylase in the Inferior Colliculus

    PubMed Central

    Bandara, Suren B.; Eubig, Paul A.; Sadowski, Renee N.; Schantz, Susan L.

    2016-01-01

    Previously, we observed that developmental polychlorinated biphenyl (PCB) exposure resulted in an increase in audiogenic seizures (AGSs) in rats. However, the rats were exposed to loud noise in adulthood, and were not tested for AGS until after 1 year of age, either of which could have interacted with early PCB exposure to increase AGS susceptibility. This study assessed susceptibility to AGS in young adult rats following developmental PCB exposure alone (without loud noise exposure) and investigated whether there was a decrease in GABA inhibitory neurotransmission in the inferior colliculus (IC) that could potentially explain this effect. Female Long-Evans rats were dosed orally with 0 or 6 mg/kg/day of an environmentally relevant PCB mixture from 28 days prior to breeding until the pups were weaned at postnatal day 21. One male-female pair from each litter was retained for the AGS study whilst another was retained for Western blot analysis of glutamic acid decarboxylase (GAD) and GABAAα1 receptor in the IC, the site in the auditory midbrain where AGS are initiated. There was a significant increase in the number and severity of AGSs in the PCB groups, with females somewhat more affected than males. GAD65 was decreased but there was no change in GAD67 or GABAAα1 in the IC indicating decreased inhibitory regulation in the PCB group. These results confirm that developmental PCB exposure alone is sufficient to increase susceptibility to AGS, and provide the first evidence for a possible mechanism of action at the level of the IC. PMID:26543103

  11. Hippocampal interneurons expressing glutamic acid decarboxylase and calcium-binding proteins decrease with aging in Fischer 344 rats.

    PubMed

    Shetty, A K; Turner, D A

    1998-05-04

    Aging leads to alterations in the function and plasticity of hippocampal circuitry in addition to behavioral changes. To identify critical alterations in the substrate for inhibitory circuitry as a function of aging, we evaluated the numbers of hippocampal interneurons that were positive for glutamic acid decarboxylase and those that expressed calcium-binding proteins (parvalbumin, calbindin, and calretinin) in young adult (4-5 months old) and aged (23-25 months old) male Fischer 344 rats. Both the overall interneuron population and specific subpopulations of interneurons demonstrated a commensurate decline in numbers throughout the hippocampus with aging. Interneurons positive for glutamic acid decarboxylase were significantly depleted in the stratum radiatum of CA1, the strata oriens, radiatum and pyramidale of CA3, the dentate molecular layer, and the dentate hilus. Parvalbumin interneurons showed significant reductions in the strata oriens and pyramidale of CA1, the stratum pyramidale of CA3, and the dentate hilus. The reductions in calbindin interneurons were more pronounced than other calcium-binding protein-positive interneurons and were highly significant in the strata oriens and radiatum of both CA1 and CA3 subfields and in the dentate hilus. Calretinin interneurons were decreased significantly in the strata oriens and radiatum of CA3, in the dentate granule cell and molecular layers, and in the dentate hilus. However, the relative ratio of parvalbumin-, calbindin-, and calretinin-positive interneurons compared with glutamic acid decarboxylase-positive interneurons remained constant with aging, suggesting actual loss of interneurons expressing calcium-binding proteins with age. This loss contrasts with the reported preservation of pyramidal neurons with aging in the hippocampus. Functional decreases in inhibitory drive throughout the hippocampus may occur due to this loss, particularly alterations in the processing of feed-forward information through the

  12. Human Monoclonal Islet Cell Antibodies From a Patient with Insulin- Dependent Diabetes Mellitus Reveal Glutamate Decarboxylase as the Target Antigen

    NASA Astrophysics Data System (ADS)

    Richter, Wiltrud; Endl, Josef; Eiermann, Thomas H.; Brandt, Michael; Kientsch-Engel, Rosemarie; Thivolet, Charles; Jungfer, Herbert; Scherbaum, Werner A.

    1992-09-01

    The autoimmune phenomena associated with destruction of the β cell in pancreatic islets and development of type 1 (insulin-dependent) diabetes mellitus (IDDM) include circulating islet cell antibodies. We have immortalized peripheral blood lymphocytes from prediabetic individuals and patients with newly diagnosed IDDM by Epstein-Barr virus transformation. IgG-positive cells were selected by anti-human IgG-coupled magnetic beads and expanded in cell culture. Supernatants were screened for cytoplasmic islet cell antibodies using the conventional indirect immunofluorescence test on cryostat sections of human pancreas. Six islet cell-specific B-cell lines, originating from a patient with newly diagnosed IDDM, could be stabilized on a monoclonal level. All six monoclonal islet cell antibodies (MICA 1-6) were of the IgG class. None of the MICA reacted with human thyroid, adrenal gland, anterior pituitary, liver, lung, stomach, and intestine tissues but all six reacted with pancreatic islets of different mammalian species and, in addition, with neurons of rat cerebellar cortex. MICA 1-6 were shown to recognize four distinct antigenic epitopes in islets. Islet cell antibody-positive diabetic sera but not normal human sera blocked the binding of the monoclonal antibodies to their target epitopes. Immunoprecipitation of 35S-labeled human islet cell extracts revealed that a protein of identical size to the enzyme glutamate decarboxylase (EC 4.1.1.15) was a target of all MICA. Furthermore, antigen immunotrapped by the MICA from brain homogenates showed glutamate decarboxylase enzyme activity. MICA 1-6 therefore reveal glutamate decarboxylase as the predominant target antigen of cytoplasmic islet cell autoantibodies in a patient with newly diagnosed IDDM.

  13. A 120nW 8b Sub-ranging SAR ADC with Signal-Dependent Charge Recycling for Biomedical Applications.

    PubMed

    Jeong, Seokhyeon; Jung, Wanyeong; Jeon, Dongsuk; Berenfeld, Omer; Oral, Hakan; Kruger, Grant; Blaauw, David; Sylvester, Dennis

    2015-06-01

    We present an 8-bit sub-ranging SAR ADC designed for bursty signals having long time periods with small code spread. A modified capacitive-DAC (CDAC) saves previous sample's MSB voltage and reuses it throughout subsequent conversions. This prevents unnecessary switching of large MSB capacitors as well as conversion cycles, reducing energy consumed in the comparator and digital logic and yielding total energy savings of 2.6×. In 0.18μm CMOS, the ADC consumes 120nW at 0.6V and 100kS/s with 46.9dB SNDR.

  14. Anti glutamate-decarboxylase antibodies: a liaison between localisation related epilepsy, stiff-person syndrome and type-1 diabetes mellitus.

    PubMed

    Szűcs, Anna; Barcs, Gábor; Winkler, Gábor; Soós, Zsuzsanna; Folyovich, András; Kelemen, Anna; Várallyay, Péter; Kamondi, Anita

    2014-07-30

    We present two patients with partial epilepsy, type-1 diabetes and stiff person syndrome associated with high serum auto-antibody levels to glutamate-decarboxylase (anti-GAD). Both patients were or have suffered from additional autoimmune conditions. The presence of stiff person syndrome and elevated anti-GAD levels have to make clinicians look for additional autoimmune conditions including type-1 diabetes. On the other hand, the co-morbidity of partial epilepsy with autoimmune conditions in patients with elevated serum anti-GAD suggests an autoimmune mechanism of partial epilepsy in these cases.

  15. Oxalate decarboxylase and oxalate oxidase activities can be interchanged with a specificity switch of up to 282,000 by mutating an active site lid.

    PubMed

    Burrell, Matthew R; Just, Victoria J; Bowater, Laura; Fairhurst, Shirley A; Requena, Laura; Lawson, David M; Bornemann, Stephen

    2007-10-30

    Oxalate decarboxylases and oxalate oxidases are members of the cupin superfamily of proteins that have many common features: a manganese ion with a common ligand set, the substrate oxalate, and dioxygen (as either a unique cofactor or a substrate). We have hypothesized that these enzymes share common catalytic steps that diverge when a carboxylate radical intermediate becomes protonated. The Bacillus subtilis decarboxylase has two manganese binding sites, and we proposed that Glu162 on a flexible lid is the site 1 general acid. We now demonstrate that a decarboxylase can be converted into an oxidase by mutating amino acids of the lid that include Glu162 with specificity switches of 282,000 (SEN161-3DAS), 275,000 (SENS161-4DSSN), and 225,000 (SENS161-4DASN). The structure of the SENS161-4DSSN mutant showed that site 2 was not affected. The requirement for substitutions other than of Glu162 was, at least in part, due to the need to decrease the Km for dioxygen for the oxidase reaction. Reversion of decarboxylase activity could be achieved by reintroducing Glu162 to the SENS161-4DASN mutant to give a relative specificity switch of 25,600. This provides compelling evidence for the crucial role of Glu162 in the decarboxylase reaction consistent with it being the general acid, for the role of the lid in controlling the Km for dioxygen, and for site 1 being the sole catalytically active site. We also report the trapping of carboxylate radicals produced during turnover of the mutant with the highest oxidase activity. Such radicals were also observed with the wild-type decarboxylase.

  16. SU-F-303-13: Initial Evaluation of Four Dimensional Diffusion- Weighted MRI (4D-DWI) and Its Effect On Apparent Diffusion Coefficient (ADC) Measurement

    SciTech Connect

    Liu, Y; Yin, F; Czito, B; Bashir, M; Palta, M; Cai, J; Zhong, X; Dale, B

    2015-06-15

    Purpose: Diffusion-weighted imaging(DWI) has been shown to have superior tumor-to-tissue contrast for cancer detection.This study aims at developing and evaluating a four dimensional DWI(4D-DWI) technique using retrospective sorting method for imaging respiratory motion for radiotherapy planning,and evaluate its effect on Apparent Diffusion Coefficient(ADC) measurement. Materials/Methods: Image acquisition was performed by repeatedly imaging a volume of interest using a multi-slice single-shot 2D-DWI sequence in the axial planes and cine MRI(served as reference) using FIESTA sequence.Each 2D-DWI image were acquired in xyz-diffusion-directions with a high b-value(b=500s/mm2).The respiratory motion was simultaneously recorded using bellows.Retrospective sorting was applied in each direction to reconstruct 4D-DWI.The technique was evaluated using a computer simulated 4D-digital human phantom(XCAT),a motion phantom and a healthy volunteer under an IRB-approved study.Motion trajectories of regions-of-interests(ROI) were extracted from 4D-DWI and compared with reference.The mean motion trajectory amplitude differences(D) between the two was calculated.To quantitatively analyze the motion artifacts,XCAT were controlled to simulate regular motion and the motions of 10 liver cancer patients.4D-DWI,free-breathing DWI(FB- DWI) were reconstructed.Tumor volume difference(VD) of each phase of 4D-DWI and FB-DWI from the input static tumor were calculated.Furthermore, ADC was measured for each phase of 4D-DWI and FB-DWI data,and mean tumor ADC values(M-ADC) were calculated.Mean M-ADC over all 4D-DWI phases was compared with M-ADC calculated from FB-DWI. Results: 4D-DWI of XCAT,the motion phantom and the healthy volunteer demonstrated the respiratory motion clearly.ROI D values were 1.9mm,1.7mm and 2.0mm,respectively.For motion artifacts analysis,XCAT 4D-DWI images show much less motion artifacts compare to FB-DWI.Mean VD for 4D-WDI and FB-DWI were 8.5±1.4% and 108±15

  17. Design of a 12-bit 2 MS/s 12 mW pipelined SAR ADC in CMOS 0.18 μm technology for CZT-based imaging system

    NASA Astrophysics Data System (ADS)

    Xue, F.; Gao, W.; Wei, X.; Liu, W.; Hu, Y.

    2016-12-01

    This paper presents a 12-bit 2 MS/s pipelined successive approximation register (SAR) ADC for CZT-based imaging system. The proposed ADC is divided into a first-stage 6-bit SAR-based Multiplying Digital Analog Converter (MDAC) and a second-stage 8-bit SAR ADC. The first-stage MDAC has a gain of 16 instead of the usual gain of 64, which considerably minimizes the power dissipation of residue amplifier. The second-stage 8-bit SAR ADC employs unit bridge capacitor split-capacitor architecture aiming to reduce the load capacitance of residue amplifier so as to minimize the power dissipation of the proposed ADC. Moreover, a code-randomized calibration algorithm is proposed to improve the linearity of the second-stage 8-bit split-capacitor SAR ADC. In addition, several radiation-hardened-by-design techniques are adopted in the layout design against space radiation effects. The prototype chip was fabricated in 0.18 μ m mixed-signal 1.8 V/3.3 V process and occupied a core area of 0.71 mm2. The proposed pipelined SAR ADC achieves a peak signal-to-noise-and-distortion ratio (SNDR) of 63.2 dB at 2 MS/s sampling rate and consumes 12 mW power in total. The figure of merit (FoM) of the proposed ADC is 5.06 pJ/conversion-step.

  18. Modifying the product pattern of Clostridium acetobutylicum: physiological effects of disrupting the acetate and acetone formation pathways.

    PubMed

    Lehmann, Dörte; Hönicke, Daniel; Ehrenreich, Armin; Schmidt, Michael; Weuster-Botz, Dirk; Bahl, Hubert; Lütke-Eversloh, Tina

    2012-05-01

    Clostridial acetone-butanol-ethanol (ABE) fermentation is a natural source for microbial n-butanol production and regained much interest in academia and industry in the past years. Due to the difficult genetic accessibility of Clostridium acetobutylicum and other solventogenic clostridia, successful metabolic engineering approaches are still rare. In this study, a set of five knock-out mutants with defects in the central fermentative metabolism were generated using the ClosTron technology, including the construction of targeted double knock-out mutants of C. acetobtuylicum ATCC 824. While disruption of the acetate biosynthetic pathway had no significant impact on the metabolite distribution, mutants with defects in the acetone pathway, including both acetoacetate decarboxylase (Adc)-negative and acetoacetyl-CoA:acyl-CoA transferase (CtfAB)-negative mutants, exhibited high amounts of acetate in the fermentation broth. Distinct butyrate increase and decrease patterns during the course of fermentations provided experimental evidence that butyrate, but not acetate, is re-assimilated via an Adc/CtfAB-independent pathway in C. acetobutylicum. Interestingly, combining the adc and ctfA mutations with a knock-out of the phosphotransacetylase (Pta)-encoding gene, acetate production was drastically reduced, resulting in an increased flux towards butyrate. Except for the Pta-negative single mutant, all mutants exhibited a significantly reduced solvent production.

  19. Inhibition of ornithine decarboxylase potentiates nitric oxide production in LPS-activated J774 cells

    PubMed Central

    Baydoun, Anwar R; Morgan, David M L

    1998-01-01

    We have examined whether modulation of the polyamine biosynthetic pathway, through inhibition by α-difluoromethylornithine (DFMO) of the rate limiting enzyme, ornithine decarboxylase (ODC), modulates NO synthesis in J774 macrophages.DFMO potentiated LPS-stimulated nitrite production in both a concentration- and time-dependent manner, increasing nitrite levels by 48±5% at 10 mM. This effect was observed in cells pre-treated with DFMO for 24 h prior to stimulation with LPS. Addition of DFMO 12 h after LPS failed to potentiate LPS-induced nitrite production.Supplementation of the culture medium with horse serum (10%) in place of foetal calf serum (10%) caused no significant change in either LPS-induced nitrite production or in the ability of DFMO (10  mM) to potentiate LPS-induced NO synthesis.Metabolism of L-[3H]arginine to L-[3H]citrulline by partially purified inducible nitric oxide synthase (iNOS) was not significantly altered by either DFMO (1–10 mM) or by putrescine (0.001–1 mM), spermidine (0.001–1 mM) or spermine (0.001–1 mM). iNOS activity was also unaffected by 1 mM EGTA but was markedly attenuated (70±0.07%) by L-NMMA (100 μM).Pre-incubation of cells with DFMO (10 mM; 24 h) prior to activation with LPS resulted in enhanced (∼2 fold) iNOS protein expression.These results show that DFMO potentiates LPS-induced nitrite production in the murine macrophage cell line J774. Since the only known mechanism of action of DFMO is inhibition of ODC, and thus polyamine biosynthesis, we conclude that expression of iNOS can be critically regulated by endogenous polyamines. PMID:9884080

  20. Rate and Equilibrium Constants for an Enzyme Conformational Change during Catalysis by Orotidine 5'-Monophosphate Decarboxylase.

    PubMed

    Goryanova, Bogdana; Goldman, Lawrence M; Ming, Shonoi; Amyes, Tina L; Gerlt, John A; Richard, John P

    2015-07-28

    The caged complex between orotidine 5'-monophosphate decarboxylase (ScOMPDC) and 5-fluoroorotidine 5'-monophosphate (FOMP) undergoes decarboxylation ∼300 times faster than the caged complex between ScOMPDC and the physiological substrate, orotidine 5'-monophosphate (OMP). Consequently, the enzyme conformational changes required to lock FOMP at a protein cage and release product 5-fluorouridine 5'-monophosphate (FUMP) are kinetically significant steps. The caged form of ScOMPDC is stabilized by interactions between the side chains from Gln215, Tyr217, and Arg235 and the substrate phosphodianion. The control of these interactions over the barrier to the binding of FOMP and the release of FUMP was probed by determining the effect of all combinations of single, double, and triple Q215A, Y217F, and R235A mutations on kcat/Km and kcat for turnover of FOMP by wild-type ScOMPDC; its values are limited by the rates of substrate binding and product release, respectively. The Q215A and Y217F mutations each result in an increase in kcat and a decrease in kcat/Km, due to a weakening of the protein-phosphodianion interactions that favor fast product release and slow substrate binding. The Q215A/R235A mutation causes a large decrease in the kinetic parameters for ScOMPDC-catalyzed decarboxylation of OMP, which are limited by the rate of the decarboxylation step, but much smaller decreases in the kinetic parameters for ScOMPDC-catalyzed decarboxylation of FOMP, which are limited by the rate of enzyme conformational changes. By contrast, the Y217A mutation results in large decreases in kcat/Km for ScOMPDC-catalyzed decarboxylation of both OMP and FOMP, because of the comparable effects of this mutation on rate-determining decarboxylation of enzyme-bound OMP and on the rate-determining enzyme conformational change for decarboxylation of FOMP. We propose that kcat = 8.2 s(-1) for decarboxylation of FOMP by the Y217A mutant is equal to the rate constant for cage formation from the

  1. Determinants of the Differential Antizyme-Binding Affinity of Ornithine Decarboxylase

    PubMed Central

    Liu, Yen-Chin; Hsu, Den-Hua; Huang, Chi-Liang; Liu, Yi-Liang; Liu, Guang-Yaw; Hung, Hui-Chih

    2011-01-01

    Ornithine decarboxylase (ODC) is a ubiquitous enzyme that is conserved in all species from bacteria to humans. Mammalian ODC is degraded by the proteasome in a ubiquitin-independent manner by direct binding to the antizyme (AZ). In contrast, Trypanosoma brucei ODC has a low binding affinity toward AZ. In this study, we identified key amino acid residues that govern the differential AZ binding affinity of human and Trypanosoma brucei ODC. Multiple sequence alignments of the ODC putative AZ-binding site highlights several key amino acid residues that are different between the human and Trypanosoma brucei ODC protein sequences, including residue 119, 124,125, 129, 136, 137 and 140 (the numbers is for human ODC). We generated a septuple human ODC mutant protein where these seven bases were mutated to match the Trypanosoma brucei ODC protein sequence. The septuple mutant protein was much less sensitive to AZ inhibition compared to the WT protein, suggesting that these amino acid residues play a role in human ODC-AZ binding. Additional experiments with sextuple mutants suggest that residue 137 plays a direct role in AZ binding, and residues 119 and 140 play secondary roles in AZ binding. The dissociation constants were also calculated to quantify the affinity of the ODC-AZ binding interaction. The Kd value for the wild type ODC protein-AZ heterodimer ([ODC_WT]-AZ) is approximately 0.22 μM, while the Kd value for the septuple mutant-AZ heterodimer ([ODC_7M]-AZ) is approximately 12.4 μM. The greater than 50-fold increase in [ODC_7M]-AZ binding affinity shows that the ODC-7M enzyme has a much lower binding affinity toward AZ. For the mutant proteins ODC_7M(-Q119H) and ODC_7M(-V137D), the Kd was 1.4 and 1.2 μM, respectively. These affinities are 6-fold higher than the WT_ODC Kd, which suggests that residues 119 and 137 play a role in AZ binding. PMID:22073206

  2. Homology-based molecular modelling of PLP-dependent histidine decarboxylase from Mmorganella morganii.

    PubMed

    Tahanejad, F S; Naderi-Manesh, H; Habibinejad, B; Mahmoudian, M

    2000-06-01

    The 3-D structural information is a prerequisite for a rational ligand design. In the absence of experimental data, model building on the basis of a known 3-D structure of a homologous protein is at present the only reliable method to obtain structural information. A homology model building study of the pyridoxal 5'-phosphate (PLP)-dependent histidine decarboxylase from Morganella morganii (HDC-MM) has been carried out based on the crystal structure of the aspartate aminotransferase from Escherichia coli (AAT-EC). The primary sequences of AAT-EC and HDC-MM were aligned by automated alignment procedure. A 3-D model of HDC-MM was constructed by copying the coordinates of the residues from the crystal structure of AAT-EC into the corresponding residues in HDC-MM. After energy-minimization of the resulting 3-D model of HDC-MM, possible active site residues were identified by fitting the substrate (l-histidine) into the proposed active-site. In our model, several residues, which have an important role in the AAT-EC active-site, are located in positions spatially identical to those in AAT-EC structure. The back-bone of the modelled active site pocket is constructed by residues; Gly-92, Gly-93, Thr-93, Ser-115, Asp-200, Ala-202, Ser-229 and Lys-232 together with residues Asn-8, His-119, Thr-171, His-198, Leu-203, His-231, Ser-236 and Ile-238. In the ligand binding site, it appears that the HDC-MM model will position l-histidine (substrate) in the area consisting of the residues; Glu-29, Ser-30, Leu-38, His-231 and Lys-232. The nitrogen atom of the imidazole ring (N2) of the substrate is predicted to interact with the carboxylate group of Ser-30. The alpha-carboxylate of histidine points toward the Lys-232 to have electrostatic interaction with its side chain nitrogen atom (N(Z)). In conclusion, this combination of sequence and 3-D structural homology between AAT-EC and HDC-MM model could provide insight in assigning the probable active site residues.

  3. Substrate Binding Mode and Molecular Basis of a Specificity Switch in Oxalate Decarboxylase

    PubMed Central

    2016-01-01

    Oxalate decarboxylase (OxDC) catalyzes the conversion of oxalate into formate and carbon dioxide in a remarkable reaction that requires manganese and dioxygen. Previous studies have shown that replacing an active-site loop segment Ser161-Glu162-Asn163-Ser164 in the N-terminal domain of OxDC with the cognate residues Asp161-Ala162-Ser-163-Asn164 of an evolutionarily related, Mn-dependent oxalate oxidase gives a chimeric variant (DASN) that exhibits significantly increased oxidase activity. The mechanistic basis for this change in activity has now been investigated using membrane inlet mass spectrometry (MIMS) and isotope effect (IE) measurements. Quantitative analysis of the reaction stoichiometry as a function of oxalate concentration, as determined by MIMS, suggests that the increased oxidase activity of the DASN OxDC variant is associated with only a small fraction of the enzyme molecules in solution. In addition, IE measurements show that C–C bond cleavage in the DASN OxDC variant proceeds via the same mechanism as in the wild-type enzyme, even though the Glu162 side chain is absent. Thus, replacement of the loop residues does not modulate the chemistry of the enzyme-bound Mn(II) ion. Taken together, these results raise the possibility that the observed oxidase activity of the DASN OxDC variant arises from an increased level of access of the solvent to the active site during catalysis, implying that the functional role of Glu162 is to control loop conformation. A 2.6 Å resolution X-ray crystal structure of a complex between oxalate and the Co(II)-substituted ΔE162 OxDC variant, in which Glu162 has been deleted from the active site loop, reveals the likely mode by which the substrate coordinates the catalytically active Mn ion prior to C–C bond cleavage. The “end-on” conformation of oxalate observed in the structure is consistent with the previously published V/K IE data and provides an empty coordination site for the dioxygen ligand that is thought to

  4. Mechanism of the Orotidine 5′-Monophosphate Decarboxylase-Catalyzed Reaction: Evidence for Substrate Destabilization

    SciTech Connect

    Chan, K.; Wood, M; Fedorov, A; Fedorov, E; Imker, H; Amyes, T; Richard, J; Almo, S; Gerlt, J

    2009-01-01

    The reaction catalyzed by orotidine 5'-monophosphate decarboxylase (OMPDC) involves a stabilized anionic intermediate, although the structural basis for the rate acceleration (kcat/knon, 7.1 x 1016) and proficiency (kcat/KM)/knon, 4.8 x 1022 M-1 is uncertain. That the OMPDCs from Methanothermobacter thermautotrophicus (MtOMPDC) and Saccharomyces cerevisiae (ScOMPDC) catalyze the exchange of H6 of the UMP product with solvent deuterium allows an estimate of a lower limit on the rate acceleration associated with stabilization of the intermediate and its flanking transition states (=1010). The origin of the 'missing' contribution, =107 (1017 total - =1010), is of interest. Based on structures of liganded complexes, unfavorable electrostatic interactions between the substrate carboxylate group and a proximal Asp (Asp 70 in MtOMPDC and Asp 91 in ScOMPDC) have been proposed to contribute to the catalytic efficiency. We investigated that hypothesis by structural and functional characterization of the D70N and D70G mutants of MtOMPDC and the D91N mutant of ScOMPDC. The substitutions for Asp 70 in MtOMPDC significantly decrease the value of kcat for decarboxylation of FOMP (a more reactive substrate analogue) but have little effect on the value of kex for exchange of H6 of FUMP with solvent deuterium; the structures of wild-type MtOMPDC and its mutants are superimposable when complexed with 6-azaUMP. In contrast, the D91N mutant of ScOMPDC does not catalyze exchange of H6 of FUMP; the structures of wild-type ScOMPDC and its D91N mutant are not superimposable when complexed with 6-azaUMP, with differences in both the conformation of the active site loop and the orientation of the ligand vis vis the active site residues. We propose that the differential effects of substitutions for Asp 70 of MtOMPDC on decarboxylation and exchange provide additional evidence for a carbanionic intermediate as well as the involvement of Asp 70 in substrate destabilization.

  5. Cholera Toxin B Subunit Linked to Glutamic Acid Decarboxylase Suppresses Dendritic Cell Maturation and Function

    PubMed Central

    Odumosu, Oludare; Nicholas, Dequina; Payne, Kimberly; Langridge, William

    2012-01-01

    Dendritic cells are the largest population of antigen presenting cells in the body. One of their main functions is to regulate the delicate balance between immunity and tolerance responsible for maintenance of immunological homeostasis. Disruption of this delicate balance often results in chronic inflammation responsible for initiation of organ specific autoimmune diseases such as rheumatoid arthritis, multiple sclerosis and type I diabetes. The cholera toxin B subunit (CTB) is a weak mucosal adjuvant known for its ability to stimulate immunity to antigenic proteins. However, conjugation of CTB to many autoantigens can induce immunological tolerance resulting in suppression of autoimmunity. In this study, we examined whether linkage of CTB to a 5 kDa C-terminal protein fragment of the major diabetes autoantigen glutamic acid decarboxylase (GAD35), can block dendritic cell (DC) functions such as biosynthesis of co-stimulatory factor proteins CD86, CD83, CD80 and CD40 and secretion of inflammatory cytokines. The results of human umbilical cord blood monocyte-derived DC - GAD35 autoantigen incubation experiments showed that inoculation of immature DCs (iDCs), with CTB-GAD35 protein dramatically suppressed levels of CD86, CD83, CD80 and CD40 co-stimulatory factor protein biosynthesis in comparison with GAD35 alone inoculated iDCs. Surprisingly, incubation of iDCs in the presence of the CTB-autoantigen and the strong immunostimulatory molecules PMA and Ionomycin revealed that CTB-GAD35 was capable of arresting PMA + Ionomycin induced DC maturation. Consistant with this finding, CTB-GAD35 mediated suppression of DC maturation was accompanied by a dramatic decrease in the secretion of the pro-inflammatory cytokines IL-12/23p40 and IL-6 and a significant increase in secretion of the immunosuppressive cytokine IL-10. Taken together, our experimental data suggest that linkage of the weak adjuvant CTB to the dominant type 1 diabetes autoantigen GAD strongly inhibits DC

  6. Biogenic Amine Degradation by Bacillus Species Isolated from Traditional Fermented Soybean Food and Detection of Decarboxylase-Related Genes.

    PubMed

    Eom, Jeong Seon; Seo, Bo Young; Choi, Hye Sun

    2015-09-01

    Biogenic amines in some food products present considerable toxicological risks as potential human carcinogens when consumed in excess concentrations. In this study, we investigated the degradation of the biogenic amines histamine and tyramine and the presence of genes encoding histidine and tyrosine decarboxylases and amine oxidase in Bacillus species isolated from fermented soybean food. No expression of histidine and tyrosine decarboxylase genes (hdc and tydc) were detected in the Bacillus species isolated (B. subtilis HJ0-6, B. subtilis D'J53-4, and B. idriensis RD13-10), although substantial levels of amine oxidase gene (yobN) expression were observed. We also found that the three selected strains, as non-biogenic amineproducing bacteria, were significantly able to degrade the biogenic amines histamine and tyramine. These results indicated that the selected Bacillus species could be used as a starter culture for the control of biogenic amine accumulation and degradation in food. Our study findings also provided the basis for the development of potential biological control agents against these biogenic amines for use in the food preservation and food safety sectors.

  7. Characterization of a pyridoxal-5'-phosphate-dependent l-lysine decarboxylase/oxidase from Burkholderia sp. AIU 395.

    PubMed

    Sugawara, Asami; Matsui, Daisuke; Takahashi, Narumi; Yamada, Miwa; Asano, Yasuhisa; Isobe, Kimiyasu

    2014-11-01

    A novel enzyme, which catalyzed decarboxylation of l-lysine into cadaverine with release of carbon dioxide and oxidative deamination of l-lysine into l-2-aminoadipic 5-semialdehyde with release of ammonia and hydrogen peroxide, was found from a newly isolated Burkholderia sp. AIU 395. The enzyme was specific to l-lysine and did not exhibit enzyme activities for other l-amino acids, l-lysine derivatives, d-amino acids, and amines. The apparent Km values for l-lysine in the oxidation and decarboxylation reactions were estimated to be 0.44 mM and 0.84 mM, respectively. The molecular mass was estimated to be 150 kDa, which was composed of two identical subunits with molecular mass of 76.5 kDa. The enzyme contained one mol of pyridoxal 5'-phosphate per subunit as a prosthetic group. The enzyme exhibiting decarboxylase and oxidase activities for l-lysine was first reported here, while the deduced amino acid sequence was homologous to that of putative lysine decarboxylases from the genus Burkholderia.

  8. Crystallization and preliminary crystallographic analysis of orotidine 5′-monophosphate decarboxylase from the human malaria parasite Plasmodium falciparum

    SciTech Connect

    Krungkrai, Sudaratana R.; Tokuoka, Keiji; Kusakari, Yukiko; Inoue, Tsuyoshi; Adachi, Hiroaki; Matsumura, Hiroyoshi; Takano, Kazufumi; Murakami, Satoshi; Mori, Yusuke; Kai, Yasushi; Krungkrai, Jerapan; Horii, Toshihiro

    2006-06-01

    Orotidine 5′-monophosphate decarboxylase of human malaria parasite P. falciparum was crystallized by the seeding method in a hanging drop using PEG 3000 as a precipitant. A complete set of diffraction data from a native crystal was collected to 2.7 Å resolution at 100 K using synchrotron radiation. Orotidine 5′-monophosphate (OMP) decarboxylase (OMPDC; EC 4.1.1.23) catalyzes the final step in the de novo synthesis of uridine 5′-monophosphate (UMP) and defects in the enzyme are lethal in the malaria parasite Plasmodium falciparum. Active recombinant P. falciparum OMPDC (PfOMPDC) was crystallized by the seeding method in a hanging drop using PEG 3000 as a precipitant. A complete set of diffraction data from a native crystal was collected to 2.7 Å resolution at 100 K using synchrotron radiation at the Swiss Light Source. The crystal exhibits trigonal symmetry (space group R3), with hexagonal unit-cell parameters a = b = 201.81, c = 44.03 Å. With a dimer in the asymmetric unit, the solvent content is 46% (V{sub M} = 2.3 Å{sup 3} Da{sup −1})

  9. Structural insights into the Escherichia coli lysine decarboxylases and molecular determinants of interaction with the AAA+ ATPase RavA

    PubMed Central

    Kandiah, Eaazhisai; Carriel, Diego; Perard, Julien; Malet, Hélène; Bacia, Maria; Liu, Kaiyin; Chan, Sze W. S.; Houry, Walid A.; Ollagnier de Choudens, Sandrine; Elsen, Sylvie; Gutsche, Irina

    2016-01-01

    The inducible lysine decarboxylase LdcI is an important enterobacterial acid stress response enzyme whereas LdcC is its close paralogue thought to play mainly a metabolic role. A unique macromolecular cage formed by two decamers of the Escherichia coli LdcI and five hexamers of the AAA+ ATPase RavA was shown to counteract acid stress under starvation. Previously, we proposed a pseudoatomic model of the LdcI-RavA cage based on its cryo-electron microscopy map and crystal structures of an inactive LdcI decamer and a RavA monomer. We now present cryo-electron microscopy 3D reconstructions of the E. coli LdcI and LdcC, and an improved map of the LdcI bound to the LARA domain of RavA, at pH optimal for their enzymatic activity. Comparison with each other and with available structures uncovers differences between LdcI and LdcC explaining why only the acid stress response enzyme is capable of binding RavA. We identify interdomain movements associated with the pH-dependent enzyme activation and with the RavA binding. Multiple sequence alignment coupled to a phylogenetic analysis reveals that certain enterobacteria exert evolutionary pressure on the lysine decarboxylase towards the cage-like assembly with RavA, implying that this complex may have an important function under particular stress conditions. PMID:27080013

  10. In vitro translation of the upstream open reading frame in the mammalian mRNA encoding S-adenosylmethionine decarboxylase.

    PubMed

    Raney, A; Baron, A C; Mize, G J; Law, G L; Morris, D R

    2000-08-11

    The upstream open reading frame (uORF) in the mRNA encoding S-adenosylmethionine decarboxylase is a polyamine-responsive element that suppresses translation of the associated downstream cistron in vivo. In this paper, we provide the first direct evidence of peptide synthesis from the S-adenosylmethionine decarboxylase uORF using an in vitro translation system. We examine both the influence of cation concentration on peptide synthesis and the effect of altering the uORF sequence on peptide synthesis. Synthesis of wild type and altered peptides was similar at all concentrations of magnesium tested. In contrast, synthesis of the wild type peptide was more sensitive than that of altered peptides to elevated concentrations of the naturally occurring polyamines, spermidine and spermine, as well as several polyamine analogs. The sensitivity of in vitro synthesis to spermidine was influenced by both the amino acid sequence and the length of the peptide product of the uORF. Findings from the present study correlate with the effects of the uORF and polyamines on translation of a downstream cistron in vivo and support the hypothesis that polyamines and the structure of the nascent peptide create a rate-limiting step in uORF translation, perhaps through a ribosome stalling mechanism.

  11. Two UDP-glucuronic acid decarboxylases involved in the biosynthesis of a bacterial exopolysaccharide in Paenibacillus elgii.

    PubMed

    Li, Ou; Qian, Chao-Dong; Zheng, Dao-Qiong; Wang, Pin-Mei; Liu, Yu; Jiang, Xin-Hang; Wu, Xue-Chang

    2015-04-01

    Xylose is described as a component of bacterial exopolysaccharides in only a limited number of bacterial strains. A bacterial strain, Paenibacillus elgii, B69 was shown to be efficient in producing a xylose-containing exopolysaccharide. Sequence analysis was performed to identify the genes encoding the uridine diphosphate (UDP)-glucuronic acid decarboxylase required for the synthesis of UDP-xylose, the precursor of the exopolysaccharide. Two sequences, designated as Peuxs1 and Peuxs2, were found as the candidate genes for such enzymes. The activities of the UDP-glucuronic acid decarboxylases were proven by heterologous expression and real-time nuclear magnetic resonance analysis. The intracellular activity and effect of these genes on the synthesis of exopolysaccharide were further investigated by developing a thymidylate synthase based knockout system. This system was used to substitute the conventional antibiotic resistance gene system in P. elgii, a natural multi-antibiotic resistant strain. Results of intracellular nucleotide sugar analysis showed that the intracellular UDP-xylose and UDP-glucuronic acid levels were affected in Peuxs1 or Peuxs2 knockout strains. The knockout of either Peuxs1 or Peuxs2 reduced the polysaccharide production and changed the monosaccharide ratio. No polysaccharide was found in the Peuxs1/Peuxs2 double knockout strain. Our results show that P. elgii can be efficient in forming UDP-xylose, which is then used for the synthesis of xylose-containing exopolysaccharide.

  12. The role of aromatic L-amino acid decarboxylase in bacillamide C biosynthesis by Bacillus atrophaeus C89.

    PubMed

    Yuwen, Lei; Zhang, Feng-Li; Chen, Qi-Hua; Lin, Shuang-Jun; Zhao, Yi-Lei; Li, Zhi-Yong

    2013-01-01

    For biosynthesis of bacillamide C by Bacillus atrophaeus C89 associated with South China sea sponge Dysidea avara, it is hypothesized that decarboxylation from L-tryptophan to tryptamine could be performed before amidation by the downstream aromatic L-amino acid decarboxylase (AADC) to the non-ribosomal peptide synthetases (NRPS) gene cluster for biosynthesizing bacillamide C. The structural analysis of decarboxylases' known substrates in KEGG database and alignment analysis of amino acid sequence of AADC have suggested that L-tryptophan and L-phenylalanine are the potential substrates of AADC. The enzymatic kinetic experiment of the recombinant AADC proved that L-tryptophan is a more reactive substrate of AADC than L-phenylalanine. Meanwhile, the AADC-catalyzed conversion of L-tryptophan into tryptamine was confirmed by means of HPLC and LC/MS. Thus during bacillamide C biosynthesis, the decarboxylation of L-tryptophan to tryptamine is likely conducted first under AADC catalysis, followed by the amidation of tryptamine with the carboxylic product of NRPS gene cluster.

  13. Proceedings of the Seventh Applied Diamond Conference/Third Frontier Carbon Technology Joint Conference (ADC/FCT 2003)

    NASA Technical Reports Server (NTRS)

    Murakawa, M. (Editor); Miyoshi, K. (Editor); Koga, Y. (Editor); Schaefer, L. (Editor); Tzeng, Y. (Editor)

    2003-01-01

    These are the Proceedings of the Seventh Applied Diamond Conference/Third Frontier Carbon Technology Joint Conference held at Epochal Tsukuba International Conference Center from August 18 to 21, 2003. The diamond CVD process was first reported by Dr. Spitsyn in 1981 and Prof. S. Iijima reported his discovery of carbon nanotubes in 1991. In the past years, both diamond-related materials and novel carbon materials have attracted considerable interest by the scientific, technological, and industrial community. Many practical and commercial products of diamond materials are reported in these proceedings. A broad variety of applications of carbon nanotubes and novel carbons have also been explored and demonstrated. Having more than 175 invited and contributing papers by authors from over 18 countries for presentations at ADC/FCT 2003 clearly demonstrates that these materials, due to the combination of their superior properties, are both scientifically amazing and economically significant.

  14. Bayes-Optimal Joint Channel-and-Data Estimation for Massive MIMO With Low-Precision ADCs

    NASA Astrophysics Data System (ADS)

    Wen, Chao-Kai; Wang, Chang-Jen; Jin, Shi; Wong, Kai-Kit; Ting, Pangan

    2016-05-01

    This paper considers a multiple-input multiple-output (MIMO) receiver with very low-precision analog-to-digital convertors (ADCs) with the goal of developing massive MIMO antenna systems that require minimal cost and power. Previous studies demonstrated that the training duration should be {\\em relatively long} to obtain acceptable channel state information. To address this requirement, we adopt a joint channel-and-data (JCD) estimation method based on Bayes-optimal inference. This method yields minimal mean square errors with respect to the channels and payload data. We develop a Bayes-optimal JCD estimator using a recent technique based on approximate message passing. We then present an analytical framework to study the theoretical performance of the estimator in the large-system limit. Simulation results confirm our analytical results, which allow the efficient evaluation of the performance of quantized massive MIMO systems and provide insights into effective system design.

  15. Proceedings of the Sixth Applied Diamond Conference/Second Frontier Carbon Technology Joint Conference (ADC/FCT 2001)

    NASA Technical Reports Server (NTRS)

    Tzeng, Y. (Editor); Miyoshi, K. (Editor); Yoshikawa, M. (Editor); Murakawa, M. (Editor); Koga, Y. (Editor); Kobashi, K. (Editor); Amaratunga, G. A. J. (Editor)

    2001-01-01

    These are the Proceedings of the Sixth Applied Diamond Conference/Second Frontier Carbon Technology Joint Conference hosted by Auburn University from August 6 to 10, 2001. The diamond CVD process was first reported by Dr. Spitsyn in 1981 and Prof. S. Iijima reported his discovery of carbon nanotubes in 1991. In the past years, both diamond-related materials and novel carbon materials have attracted considerable interest by the scientific, technological, and industrial community. Many practical and commercial products of diamond materials are reported in these proceedings. A broad variety of applications of carbon nanotubes and novel carbons have also been explored and demonstrated. Having more than 200 invited and contributing papers by authors from over 20 countries for presentations at ADC/FCT 2001 clearly demonstrates that these materials, due to the combination of their superior properties, are both scientifically amazing and economically significant.

  16. Parametric Response Mapping of Apparent Diffusion Coefficient (ADC) as an Imaging Biomarker to Distinguish Pseudoprogression from True Tumor Progression In Peptide-Based Vaccine Therapy for Pediatric Diffuse Instrinsic Pontine Glioma

    PubMed Central

    Ceschin, Rafael; Kurland, Brenda F.; Abberbock, Shira R.; Ellingson, Benjamin M.; Okada, Hideho; Jakacki, Regina I.; Pollack, Ian F.; Panigrahy, Ashok

    2015-01-01

    Background and Purpose Immune response to cancer therapy may result in pseudoprogression, which can only be identified retrospectively and which may disrupt an effective therapy. This study assesses whether serial parametric response mapping (PRM, a voxel-by-voxel method of image analysis also known as functional diffusion mapping) analysis of ADC measurements following peptide-based vaccination may help prospectively distinguish progression from pseudoprogression in pediatric patients with diffuse intrinsic pontine gliomas. Materials and Methods From 2009–2012, 21 children age 4–18 with diffuse intrinsic pontine gliomas were enrolled in a serial peptide-based vaccination protocol following radiotherapy. DWI was acquired before immunotherapy and at six week intervals during vaccine treatment. Pseudoprogression was identified retrospectively based on clinical and radiographic findings, excluding DWI. Parametric response mapping was used to analyze 96 scans, comparing ADC measures at multiple time points (from first vaccine to up to 12 weeks after the vaccine was halted) to pre-vaccine baseline values. Log-transformed fractional increased ADC (fiADC), fractional decreased ADC (fdADC), and parametric response mapping ratio (fiADC/fdADC) were compared between patients with and without pseudoprogression, using generalized estimating equations with inverse weighting by cluster size. Results Median survival was 13.1 months from diagnosis (range 6.4–24.9 months). Four of 21 children (19%) were assessed as experiencing pseudoprogression. Patients with pseudoprogression had higher fitted average log-transformed parametric response mapping ratios (p=0.01) and fiADCs (p=0.0004), compared to patients without pseudoprogression. Conclusion Serial parametric response mapping of ADC, performed at multiple time points of therapy, may distinguish pseudoprogression from true progression in patients with diffuse intrinsic pontine gliomas treated with peptide-based vaccination

  17. A low power front-end architecture for SiPM readout with integrated ADC and multiplexed readout

    NASA Astrophysics Data System (ADS)

    Sacco, I.; Fischer, P.; Ritzert, M.; Peric, I.

    2013-01-01

    Silicon Photo-Multiplier (SiPM) detectors are becoming widely used for optical photon and, in conjunction with suited scintillators, for gamma detection in both medical imaging and particle physics experiments. The spatial resolution can be improved by using smaller SiPMs with a corresponding increase in front-end channels density. The timing resolution of the whole system is a function of the detector parameters and of the characteristics of the front-end electronics. We present a low power front-end readout architecture which allows reading out several SiPMs though a single line in order to maximize the number of SiPMs. The design offers good timing performance and includes a simple charge digitizer in every channel. Four different single-ended channel designs have been designed, submitted for fabrication and characterized electronically and with SiPMs. The timing performance is obtained by using a low input impedance, precise threshold setting of a leading edge discriminator and a programmable input dc potential to set the SiPM HV bias on a channel per channel basis. Programmable low- and high-pass filters should allow reducing baseline fluctuations and noise. A simple ADC is implemented by first integrating the signal current and then discharging it at a constant rate until the baseline is reached again. The current consumption of the single channel is typically less than 10 mA. The time and energy information are sent out on a single wire. In order to keep as low as possible the output cabling the signals from different channels can be multiplexed on the same cable. The processing of these signals (extraction of time, ADC amplitude determination and channel number decoding) is performed by an external FPGA. The overall architecture, the front-end designs, and measurements with SiPMs are presented.

  18. A readout integrated circuit based on DBI-CTIA and cyclic ADC for MEMS-array-based focal plane

    NASA Astrophysics Data System (ADS)

    Miao, Liu; Dong, Wu; Zheyao, Wang

    2016-11-01

    A readout integrated circuit (ROIC) for a MEMS (microelectromechanical system)-array-based focal plane (MAFP) intended for imaging applications is presented. The ROIC incorporates current sources for diode detectors, scanners, timing sequence controllers, differential buffered injection-capacitive trans-impedance amplifier (DBI-CTIA) and 10-bit cyclic ADCs, and is integrated with MAFP using 3-D integration technology. A small-signal equivalent model is built to include thermal detectors into circuit simulations. The biasing current is optimized in terms of signal-to-noise ratio and power consumption. Layout design is tailored to fulfill the requirements of 3-D integration and to adapt to the size of MAFP elements, with not all but only the 2 bottom metal layers to complete nearly all the interconnections in DBI-CTIA and ADC in a 40 μm wide column. Experimental chips are designed and fabricated in a 0.35 μm CMOS mixed signal process, and verified in a code density test of which the results indicate a (0.29/-0.31) LSB differential nonlinearity (DNL) and a (0.61/-0.45) LSB integral nonlinearity (INL). Spectrum analysis shows that the effective number of bits (ENOB) is 9.09. The ROIC consumes 248 mW of power at most if not to cut off quiescent current paths when not needed. Project supported by by National Natural Science Foundation of China (No. 61271130), the Beijing Municipal Science and Tech Project (No. D13110100290000), the Tsinghua University Initiative Scientific Research Program (No. 20131089225), and the Shenzhen Science and Technology Development Fund (No. CXZZ20130322170740736).

  19. A modified pathway for the production of acetone in Escherichia coli.

    PubMed

    May, Antje; Fischer, Ralf-Jörg; Maria Thum, Simone; Schaffer, Steffen; Verseck, Stefan; Dürre, Peter; Bahl, Hubert

    2013-01-01

    A modified synthetic acetone operon was constructed. It consists of two genes from Clostridium acetobutylicum (thlA coding for thiolase and adc coding for acetoacetate decarboxylase) and one from Bacillus subtilis or Haemophilus influenzae (teII(srf) or ybgC, respectively, for thioesterase). Expression of this operon in Escherichia coli resulted in the production of acetone starting from the common metabolite acetyl-CoA via acetoacetyl-CoA and acetoacetate. The thioesterases do not need a CoA acceptor for acetoacetyl-CoA hydrolysis. Thus, in contrast to the classic acetone pathway of Clostridium acetobutylicum and related microorganisms which employ a CoA transferase, the new pathway is acetate independent. The genetic background of the host strains was crucial. Only E. coli strains HB101 and WL3 were able to produce acetone via the modified plasmid based pathway, up to 64mM and 42mM in 5-ml cultures, respectively. Using glucose fed-batch cultures the concentration could be increased up to 122mM acetone with HB101 carrying the recombinant plasmid pUC19ayt (thioesterase from H. influenzae). The formation of acetone led to a decreased acetate production by E. coli.

  20. Gd-EOB-DTPA-Enhanced MR Imaging of the Liver: The Effect on T2 Relaxation Times and Apparent Diffusion Coefficient (ADC)

    PubMed Central

    Cieszanowski, Andrzej; Podgórska, Joanna; Rosiak, Grzegorz; Maj, Edyta; Grudziński, Ireneusz P.; Kaczyński, Bartosz; Szeszkowski, Wojciech; Milczarek, Krzysztof; Rowiński, Olgierd

    2016-01-01

    Summary Background To investigate the effect of gadoxetic acid disodium (Gd-EOB-DTPA) on T2 relaxation times and apparent diffusion coefficient (ADC) values of the liver and focal liver lesions on a 1.5-T system. Material/Methods Magnetic resonance (MR) studies of 50 patients with 35 liver lesions were retrospectively analyzed. All examinations were performed at 1.5T and included T2-weighted turbo spin-echo (TSE) and diffusion-weighted (DW) images acquired before and after intravenous administration of Gd-EOB-DTPA. To assess the effect of this hepatobiliary contrast agent on T2-weighted TSE images and DW images T2 relaxation times and ADC values of the liver and FLLs were calculated and compared pre- and post-injection. Results The mean T2 relaxation times of the liver and focal hepatic lesions were lower on enhanced than on unenhanced T2-weighted TSE images (decrease of 2.7% and 3.6% respectively), although these differences were not statistically significant. The mean ADC values of the liver showed statistically significant decrease (of 4.6%) on contrast-enhanced DW images, compared to unenhanced images (P>0.05). The mean ADC value of liver lesions was lower on enhanced than on unenhanced DW images, but this difference (of 2.9%) did not reach statistical significance. Conclusions The mean T2 relaxation times of the liver and focal liver lesions as well as the mean ADC values of liver lesions were not significantly different before and after administration of Gd-EOB-DTPA. Therefore, acquisition of T2-weighted and DW images between the dynamic contrast-enhanced examination and hepatobiliary phase is feasible and time-saving. PMID:27026795

  1. Imipenem-resistant Acinetobacter baumannii carrying the ISAba1-bla OXA-23,51 and ISAba1-bla ADC-7 genes in Monteria, Colombia.

    PubMed

    Martínez, Pedro; Mattar, Salim

    2012-10-01

    The purpose of this study was to identify the genes coding for resistance to ceftazidime and imipenem and describe the molecular epidemiology of A. baumannii strains isolated from a clinical center in Colombia. Twenty isolates of imipenem-resistant A. baumannii from an equal number of patients with nosocomial infections were obtained. Primers were used to amplify genes bla IMP, bla VIM, bla OXA-23, bla OXA-24, bla OXA-58, bla OXA-51 and bla ADC-7. To detect insertion sequences ISAba1/bla OXA-23, ISAba1/bla OXA-51 and ISAba1/bla ADC-7, mapping by PCR using combinations of reverse primers ISAba1 and reverse primers of bla OXA-23, bla OXA-51 and bla ADC-7 were used. The amplification products were purified and cloned into PCR 2.1-TOPO vector and transformed into chemically competent Escherichia coli TOP10. These amplicons were then sequenced. PFGE was performed on DNA of A. baumannii isolates digested with ApaI. Results. The DNA profiles obtained included 9 clusters with, four 2-7 isolates per profile, and 5 single-isolate profiles. Of the 20 isolates resistant to imipenem, 15 carried bla OXA-23 gene, 4 contained ISAba1 upstream of bla OXA-51 gene, and 6 contained ISAba1 upstream of bla OXA-23 gene. Eighteen of these isolates carried the bla ADC-7 gene, with 9 of the isolates having ISAba1 located upstream of this gene. This is the first report of the ISAba1/ADC-7 associated with OXAs genes in A. baumannii isolates from Colombia.

  2. Detection and Localization of Prostate Cancer With the Targeted Biopsy Strategy Based on ADC Map: A Prospective Large-Scale Cohort Study

    PubMed Central

    Watanabe, Yuji; Terai, Akito; Araki, Tohru; Nagayama, Masako; Okumura, Akira; Amoh, Yoshiki; Ishimori, Takayoshi; Ishibashi, Mana; Nakashita, Satoru; Dodo, Yoshihiro

    2012-01-01

    Purpose To investigate the usefulness of targeted biopsy strategy based on apparent diffusion coefficient (ADC) maps in the detection and localization of prostate cancer. Materials and Methods Institutional review board approval and informed consent from all participants were obtained. This study included 1448 consecutive patients suspected of having prostate cancer based on PSA level, who were divided into two groups: Group A included 890 patients with low-ADC lesions who underwent targeted and systematic biopsies; Group B included 558 patients with no low-ADC lesions who underwent only systematic biopsies. The cancer detection rates (CDR) of each group, positive predictive value (PPV), and negative predictive value (NPV) of ADC maps were calculated. Results The CDR was 70.1% for Group A, higher than those for overall patients (48.1%) and for Group B (13.1%) with significant difference (P < 0.001). In the serum, PSA range from 4 to 20 ng/mL, the CDR was higher for the Group A than for the Group B and overall patients with significant differences. PPV and NPV of MR findings were 70.1% and 86.9%, respectively. Especially, the PPV of the MR findings for the anterior portion was as high as 90.1%. Among the false negatives of MR findings, Gleason score proved 6 or smaller in 79.5%, and positive core number was merely one or two in 80.8%. Conclusion The targeted biopsy strategy based on ADC maps can be useful in the detection and localization of prostate cancer with high PPV. J. Magn. Reson. Imaging 2012;35:1414–1421. © 2012 Wiley Periodicals, Inc. PMID:22246980

  3. Correlation of (18)F-FDG PET and MR Apparent Diffusion Coefficient (ADC) Histogram Metrics with Survival in Diffuse Intrinsic Pontine Glioma: A Report from the Pediatric Brain Tumor Consortium.

    PubMed

    Zukotynski, Katherine; Vajapeyam, Sridhar; Fahey, Frederic H; Kocak, Mehmet; Brown, Douglas; Ricci, Kelsey; Onar-Thomas, Arzu; Fouladi, Maryam; Poussaint, Tina Young

    2017-03-30

    Rationale: To describe baseline (18)F-labeled 2-fluoro-2-deoxy-D-glucose positron emission tomography (FDG-PET) voxel characteristics in pediatric diffuse intrinsic pontine glioma (DIPG) and to correlate these metrics with baseline magnetic resonance (MR) apparent diffusion coefficient (ADC) histogram metrics, progression-free survival (PFS) and overall survival (OS). Methods: Baseline brain FDG-PET and MR scans were obtained in 33 children from Pediatric Brain Tumor Consortium (PBTC) clinical DIPG trials. FDG-PET, post-gadolinium (PG) and ADC images were registered to baseline fluid attenuation inversion recovery (FLAIR) images. Three-dimensional regions of interest on FLAIR and PG images and FDG-PET and ADC histograms were generated. Metrics evaluated included peak number, skewness and kurtosis. Correlation between PET and ADC histogram metrics was evaluated. PET pixel values within the ROI for each tumor were plotted against ADC values. Association of these imaging markers with survival was described. Results: PET histograms were almost always unimodal (94% vs. 6% bimodal). None of the PET histogram parameters (skewness or kurtosis) had a significant association with PFS, although a higher PET PG skewness tended towards less favorable PFS (Hazard Ratio (95% CI)=3.48 (0.75, 16.28); P = 0.11). There was a significant association of higher ADC PG skewness with shorter PFS (Hazard Ratio (95% CI)=2.56 (1.11, 5.91); P = 0.028) and the suggestion that this also led to shorter OS (Hazard Ratio (95% CI)=2.18 (0.95, 5.04); P = 0.067). Higher ADC PG kurtosis tended towards shorter PFS (Hazard Ratio (95% CI)=1.30 (0.98, 1.74); P = 0.073). In a number of cases, PET and ADC pixel values were negatively correlated using the Pearson correlation coefficient. Further, the level of PET and ADC correlation was significantly positively associated with PFS; tumors with higher values of ADC-PET correlation had more favorable PFS (Hazard Ratio (95% CI)=0.17 (0.03, 0.89), P = 0

  4. Identification of a tyrosine decarboxylase gene (tdcA) in Streptococcus thermophilus 1TT45 and analysis of its expression and tyramine production in milk.

    PubMed

    La Gioia, Federica; Rizzotti, Lucia; Rossi, Franca; Gardini, Fausto; Tabanelli, Giulia; Torriani, Sandra

    2011-02-01

    In this study, a tyrosine decarboxylase gene (tdcA) was identified in 1 among 83 Streptococcus thermophilus strains tested. Its sequence, nearly identical to that of a tdcA of Lactobacillus curvatus, indicated a horizontal gene transfer event. Transcription in milk and the formation of critical levels of tyramine were observed in the presence of tyrosine.

  5. Stimulation of L-asparate beta-decarboxylase formation by L-glutamate in Pseudomonas dacunhae and Improved production of L-alanine.

    PubMed

    Shibatani, T; Kakimoto, T; Chibata, I

    1979-09-01

    The formation of L-asparate beta-decarboxylase by Pseudomonas dacunhae was compared on media containing a variety of organic acids and amino acids as a carbon source. Although the enzyme was formed constitutively when the organism was grown on basal medium or on that containing tricarboxylic acid cycle intermediates, it was induced twofold by L-glutamate and repressed one-tenth by L-serine. L-Glutamine, L-proline, L-leucine, glycine, and L-threonine also showed induction effects lower than that of L-glutamate. L-Glutamate derepressed the serine effect. This glutamate effect was observed effect was observed with other microoganisms, e.g., Achromobacter pestifer and Achromobacter liquidum. Since the intermediates from L-glutamate metabolism had no effect, this induction effect was specific to L-glutamate. The formation of some glutamate-related enzymes was measured and is discussed in relation to the formation of L-asparate beta-decarboxylase. L-Asparate beta-decarboxylase was purified to an electrophoretically homogenous state from L-glutamate-grown cells of P. dacunhae, and some properties were compared with those of the enzyme from fumarate-grown cells. The two enzymes were identical in disc electrophoresis, molecular weight, and some enzymatic properties. The industrial production of L-alanine from L-aspartic acid acid was improved by using the culture broth with highly induced L-asparate beta-decarboxylase (9.4 U/ml of broth).

  6. Pyruvate Decarboxylase Catalyzes Decarboxylation of Branched-Chain 2-Oxo Acids but Is Not Essential for Fusel Alcohol Production by Saccharomyces cerevisiae

    PubMed Central

    ter Schure, Eelko G.; Flikweert, Marcel T.; van Dijken, Johannes P.; Pronk, Jack T.; Verrips, C. Theo

    1998-01-01

    The fusel alcohols 3-methyl-1-butanol, 2-methyl-1-butanol, and 2-methyl-propanol are important flavor compounds in yeast-derived food products and beverages. The formation of these compounds from branched-chain amino acids is generally assumed to occur via the Ehrlich pathway, which involves the concerted action of a branched-chain transaminase, a decarboxylase, and an alcohol dehydrogenase. Partially purified preparations of pyruvate decarboxylase (EC 4.1.1.1) have been reported to catalyze the decarboxylation of the branched-chain 2-oxo acids formed upon transamination of leucine, isoleucine, and valine. Indeed, in a coupled enzymatic assay with horse liver alcohol dehydrogenase, cell extracts of a wild-type Saccharomyces cerevisiae strain exhibited significant decarboxylation rates with these branched-chain 2-oxo acids. Decarboxylation of branched-chain 2-oxo acids was not detectable in cell extracts of an isogenic strain in which all three PDC genes had been disrupted. Experiments with cell extracts from S. cerevisiae mutants expressing a single PDC gene demonstrated that both PDC1- and PDC5-encoded isoenzymes can decarboxylate branched-chain 2-oxo acids. To investigate whether pyruvate decarboxylase is essential for fusel alcohol production by whole cells, wild-type S. cerevisiae and an isogenic pyruvate decarboxylase-negative strain were grown on ethanol with a mixture of leucine, isoleucine, and valine as the nitrogen source. Surprisingly, the three corresponding fusel alcohols were produced in both strains. This result proves that decarboxylation of branched-chain 2-oxo acids via pyruvate decarboxylase is not an essential step in fusel alcohol production. PMID:9546164

  7. The ferredoxin-like domain of the activating enzyme is required for generating a lasting glycyl radical in 4-hydroxyphenylacetate decarboxylase.

    PubMed

    Selvaraj, Brinda; Pierik, Antonio J; Bill, Eckhard; Martins, Berta M

    2014-12-01

    4-Hydroxyphenylacetate decarboxylase-activating enzyme (4Hpad-AE) uses S-adenosylmethionine (SAM or AdoMet) and a [4Fe-4S] ²⁺/⁺cluster (RS cluster) to generate a stable glycyl radical on the decarboxylase. 4Hpad-AE might bind up to two auxiliary [4Fe-4S] clusters coordinated by a ferredoxin-like insert C-terminal to the RS cluster-binding motif. Except for the AEs of pyruvate formate-lyase and anaerobic ribonucleotide reductase, all glycyl radical-activating enzymes possess a similar ferredoxin-like domain, whose functional role is still poorly understood. To assess the role of the putative ferredoxin clusters from 4Hpad-AE, we combined biochemical and spectroscopic methods to characterize a truncated version of the protein (Δ66-AE) devoid of the ferredoxin-like domain. We found that Δ66-AE is stable, harbors a fully active RS cluster and can activate the decarboxylase. From the similar cleavage rates for S-adenosylmethionine of Δ66-AE and wild-type AE, we infer the reactivity of the RS cluster is unperturbed by the absence of the ferredoxin-like domain. Thus, the auxiliary clusters are not required as electron conduit to the RS cluster for effective reductive cleavage of SAM. The activation of the decarboxylase by Δ66-AE is almost as fast as with wild-type AE, but the generated glycyl radical is short living. We postulate that the ferredoxin-like domain is not required for SAM-dependent glycyl radical generation in the decarboxylase, but is necessary for producing a lasting glycyl radical.

  8. Crystallization and preliminary crystallographic analysis of orotidine 5′-monophosphate decarboxylase from the human malaria parasite Plasmodium falciparum

    PubMed Central

    Krungkrai, Sudaratana R.; Tokuoka, Keiji; Kusakari, Yukiko; Inoue, Tsuyoshi; Adachi, Hiroaki; Matsumura, Hiroyoshi; Takano, Kazufumi; Murakami, Satoshi; Mori, Yusuke; Kai, Yasushi; Krungkrai, Jerapan; Horii, Toshihiro

    2006-01-01

    Orotidine 5′-monophosphate (OMP) decarboxylase (OMPDC; EC 4.1.1.23) catalyzes the final step in the de novo synthesis of uridine 5′-monophosphate (UMP) and defects in the enzyme are lethal in the malaria parasite Plasmodium falciparum. Active recombinant P. falciparum OMPDC (PfOMPDC) was crystallized by the seeding method in a hanging drop using PEG 3000 as a precipitant. A complete set of diffraction data from a native crystal was collected to 2.7 Å resolution at 100 K using synchrotron radiation at the Swiss Light Source. The crystal exhibits trigonal symmetry (space group R3), with hexagonal unit-cell parameters a = b = 201.81, c = 44.03 Å. With a dimer in the asymmetric unit, the solvent content is 46% (V M = 2.3 Å3 Da−1). PMID:16754976

  9. Atomic Resolution Structure of the Orotidine 5′-Monophosphate Decarboxylase Product Complex Combined with Surface Plasmon Resonance Analysis

    PubMed Central

    Fujihashi, Masahiro; Mito, Kazuya; Pai, Emil F.; Miki, Kunio

    2013-01-01

    Orotidine 5′-monophosphate decarboxylase (ODCase) accelerates the decarboxylation of its substrate by 17 orders of magnitude. One argument brought forward against steric/electrostatic repulsion causing substrate distortion at the carboxylate substituent as part of the catalysis has been the weak binding affinity of the decarboxylated product (UMP). The crystal structure of the UMP complex of ODCase at atomic resolution (1.03 Å) shows steric competition between the product UMP and the side chain of a catalytic lysine residue. Surface plasmon resonance analysis indicates that UMP binds 5 orders of magnitude more tightly to a mutant in which the interfering side chain has been removed than to wild-type ODCase. These results explain the low affinity of UMP and counter a seemingly very strong argument against a contribution of substrate distortion to the catalytic reaction mechanism of ODCase. PMID:23395822

  10. Structure of lpg0406, a carboxymuconolactone decarboxylase family protein possibly involved in antioxidative response from Legionella pneumophila.

    PubMed

    Chen, Xiaofang; Hu, Yanjin; Yang, Bo; Gong, Xiaojian; Zhang, Nannan; Niu, Liwen; Wu, Yun; Ge, Honghua

    2015-12-01

    Lpg0406, a hypothetical protein from Legionella pneumophila, belongs to carboxymuconolactone decarboxylase (CMD) family. We determined the crystal structure of lpg0406 both in its apo and reduced form. The structures reveal that lpg0406 forms a hexamer and have disulfide exchange properties. The protein has an all-helical fold with a conserved thioredoxin-like active site CXXC motif and a proton relay system similar to that of alkylhydroperoxidase from Mycobacterium tuberculosis (MtAhpD), suggesting that lpg0406 might function as an enzyme with peroxidase activity and involved in antioxidant defense. A comparison of the size and the surface topology of the putative substrate-binding region between lpg0406 and MtAhpD indicates that the two enzymes accommodate the different substrate preferences. The structural findings will enhance understanding of the CMD family protein structure and its various functions.

  11. Amino acids allosterically regulate the thiamine diphosphate-dependent alpha-keto acid decarboxylase from Mycobacterium tuberculosis.

    PubMed

    Werther, Tobias; Spinka, Michael; Tittmann, Kai; Schütz, Anja; Golbik, Ralph; Mrestani-Klaus, Carmen; Hübner, Gerhard; König, Stephan

    2008-02-29

    The gene rv0853c from Mycobacterium tuberculosis strain H37Rv codes for a thiamine diphosphate-dependent alpha-keto acid decarboxylase (MtKDC), an enzyme involved in the amino acid degradation via the Ehrlich pathway. Steady state kinetic experiments were performed to determine the substrate specificity of MtKDC. The mycobacterial enzyme was found to convert a broad spectrum of branched-chain and aromatic alpha-keto acids. Stopped-flow kinetics showed that MtKDC is allosterically activated by alpha-keto acids. Even more, we demonstrate that also amino acids are potent activators of this thiamine diphosphate-dependent enzyme. Thus, metabolic flow through the Ehrlich pathway can be directly regulated at the decarboxylation step. The influence of amino acids on MtKDC catalysis was investigated, and implications for other thiamine diphosphate-dependent enzymes are discussed.

  12. Improvement of ethanol production by recombinant expression of pyruvate decarboxylase in the white-rot fungus Phanerochaete sordida YK-624.

    PubMed

    Wang, Jianqiao; Hirabayashi, Sho; Mori, Toshio; Kawagishi, Hirokazu; Hirai, Hirofumi

    2016-07-01

    To improve ethanol production by Phanerochaete sordida YK-624, the pyruvate decarboxylase (PDC) gene was cloned from and reintroduced into this hyper lignin-degrading fungus; the gene encodes a key enzyme in alcoholic fermentation. We screened 16 transformant P. sordida YK-624 strains that each expressed a second, recombinant PDC gene (pdc) and then identified the transformant strain (designated GP7) with the highest ethanol production. Direct ethanol production from hardwood was 1.41 higher with GP7 than with wild-type P. sordida YK-624. RT-PCR analysis indicated that the increased PDC activity was caused by elevated recombinant pdc expression. Taken together, these results suggested that ethanol production by P. sordida YK-624 can be improved by the stable expression of an additional, recombinant pdc.

  13. Inhibition of placental ornithine decarboxylase by DL-alpha-difluoro-methyl ornithine causes fetal growth restriction in rat.

    PubMed

    Ishida, Makoto; Hiramatsu, Yuji; Masuyama, Hisashi; Mizutani, Yasushi; Kudo, Takafumi

    2002-02-08

    The roles of polyamines in intrauterine growth restriction (IUGR) is studied. The DL-alpha-difluoromethyl ornithine (DFMO), an irreversible inhibitor of ornithine decarboxylase (ODC) which is a rate limiting enzyme of polyamine synthesis was administrated to pregnant rats so that we obtained rat fetuses with IUGR. The changes of maternal nutrition, damage of the placenta, and the direct effect of DFMO on the fetus were examined in this IUGR model. Administration of DFMO did not induced changes of maternal nutrition except for triglyceride and the fetal metabolic state. But the placental weight, ODC activity, and DNA in the placenta were decreased significantly. The ODC activity in the total placenta decreased to less than 10% of that of the control. Depression of ODC activity in the placenta may be the major cause of IUGR induced by DFMO administration, and polyamines play important roles to carry pregnancy.

  14. Ornithine and arginine decarboxylase activities and effect of some polyamine biosynthesis inhibitors on Gigaspora rosea germinating spores.

    PubMed

    Sannazzaro, Analía I; Alvarez, Cora L; Menéndez, Ana B; Pieckenstain, Fernando L; Albertó, Edgardo O; Ruiz, Oscar A

    2004-01-15

    The pathways for putrescine biosynthesis and the effects of polyamine biosynthesis inhibitors on the germination and hyphal development of Gigaspora rosea spores were investigated. Incubation of spores with different radioactive substrates demonstrated that both arginine and ornithine decarboxylase pathways participate in putrescine biosynthesis in G. rosea. Spermidine and spermine were the most abundant polyamines in this fungus. The putrescine biosynthesis inhibitors alpha-difluoromethylarginine and alpha-difluoromethylornithine, as well as the spermidine synthase inhibitor cyclohexylamine, slightly decreased polyamine levels. However, only the latter interfered with spore germination. The consequences of the use of putrescine biosynthesis inhibitors for the control of plant pathogenic fungi on the viability of G. rosea spores in soil are discussed.

  15. Inhibition of Morganella morganii Histidine Decarboxylase Activity and Histamine Accumulation in Mackerel Muscle Derived from Filipendula ulumaria Extracts.

    PubMed

    Nitta, Yoko; Yasukata, Fumiko; Kitamoto, Noritoshi; Ito, Mikiko; Sakaue, Motoyoshi; Kikuzaki, Hiroe; Ueno, Hiroshi

    2016-03-01

    Filipendula ulmaria, also known as meadowsweet, is an herb; its extract was examined for the prevention of histamine production, primarily that caused by contaminated fish. The efficacy of meadowsweet was assessed using two parameters: inhibition of Morganella morganii histidine decarboxylase (HDC) and inhibition of histamine accumulation in mackerel. Ellagitannins from F. ulmaria (rugosin D, rugosin A methyl ester, tellimagrandin II, and rugosin A) were previously shown to be potent inhibitors of human HDC; and in the present work, these compounds inhibited M. morganii HDC, with half maximal inhibitory concentration values of 1.5, 4.4, 6.1, and 6.8 μM, respectively. Application of the extracts (at 2 wt%) to mackerel meat yielded significantly decreased histamine accumulation compared with treatment with phosphate-buffered saline as a control. Hence, F. ulmaria exhibits inhibitory activity against bacterial HDC and might be effective for preventing food poisoning caused by histamine.

  16. Free-running ADC- and FPGA-based signal processing method for brain PET using GAPD arrays

    NASA Astrophysics Data System (ADS)

    Hu, Wei; Choi, Yong; Hong, Key Jo; Kang, Jihoon; Jung, Jin Ho; Huh, Youn Suk; Lim, Hyun Keong; Kim, Sang Su; Kim, Byung-Tae; Chung, Yonghyun

    2012-02-01

    Currently, for most photomultiplier tube (PMT)-based PET systems, constant fraction discriminators (CFD) and time to digital converters (TDC) have been employed to detect gamma ray signal arrival time, whereas anger logic circuits and peak detection analog-to-digital converters (ADCs) have been implemented to acquire position and energy information of detected events. As compared to PMT the Geiger-mode avalanche photodiodes (GAPDs) have a variety of advantages, such as compactness, low bias voltage requirement and MRI compatibility. Furthermore, the individual read-out method using a GAPD array coupled 1:1 with an array scintillator can provide better image uniformity than can be achieved using PMT and anger logic circuits. Recently, a brain PET using 72 GAPD arrays (4×4 array, pixel size: 3 mm×3 mm) coupled 1:1 with LYSO scintillators (4×4 array, pixel size: 3 mm×3 mm×20 mm) has been developed for simultaneous PET/MRI imaging in our laboratory. Eighteen 64:1 position decoder circuits (PDCs) were used to reduce GAPD channel number and three off-the-shelf free-running ADC and field programmable gate array (FPGA) combined data acquisition (DAQ) cards were used for data acquisition and processing. In this study, a free-running ADC- and FPGA-based signal processing method was developed for the detection of gamma ray signal arrival time, energy and position information all together for each GAPD channel. For the method developed herein, three DAQ cards continuously acquired 18 channels of pre-amplified analog gamma ray signals and 108-bit digital addresses from 18 PDCs. In the FPGA, the digitized gamma ray pulses and digital addresses were processed to generate data packages containing pulse arrival time, baseline value, energy value and GAPD channel ID. Finally, these data packages were saved to a 128 Mbyte on-board synchronous dynamic random access memory (SDRAM) and then transferred to a host computer for coincidence sorting and image reconstruction. In order to

  17. Trypanosoma cruzi has not lost its S-adenosylmethionine decarboxylase: characterization of the gene and the encoded enzyme.

    PubMed Central

    Persson, K; Aslund, L; Grahn, B; Hanke, J; Heby, O

    1998-01-01

    All attempts to identify ornithine decarboxylase in the human pathogen Trypanosoma cruzi have failed. The parasites have instead been assumed to depend on putrescine uptake and S-adenosylmethionine decarboxylase (AdoMetDC) for their synthesis of the polyamines spermidine and spermine. We have now identified the gene encoding AdoMetDC in T. cruzi by PCR cloning, with degenerate primers corresponding to conserved amino acid sequences in AdoMetDC proteins of other trypanosomatids. The amplified DNA fragment was used as a probe to isolate the complete AdoMetDC gene from a T. cruzi genomic library. The AdoMetDC gene was located on chromosomes with a size of approx. 1.4 Mbp, and contained a coding region of 1110 bp, specifying a sequence of 370 amino acid residues. The protein showed a sequence identity of only 25% with human AdoMetDC, the major differences being additional amino acids present in the terminal regions of the T. cruzi enzyme. As expected, a higher sequence identity (68-72%) was found in comparison with trypanosomatid AdoMetDCs. When the coding region was expressed in Escherichia coli, the recombinant protein underwent autocatalytic cleavage, generating a 33-34 kDa alpha subunit and a 9 kDa beta subunit. The encoded protein catalysed the decarboxylation of AdoMet (Km 0.21 mM) and was stimulated by putrescine but inhibited by the polyamines, weakly by spermidine and strongly by spermine. Methylglyoxal-bis(guanylhydrazone) (MGBG), a potent inhibitor of human AdoMetDC, was a poor inhibitor of the T. cruzi enzyme. This differential sensitivity to MGBG suggests that the two enzymes are sufficiently different to warrant the search for compounds that might interfere with the progression of Chagas' disease by selectively inhibiting T. cruzi AdoMetDC. PMID:9677309

  18. Aspartate 203 of the oxaloacetate decarboxylase beta-subunit catalyses both the chemical and vectorial reaction of the Na+ pump.

    PubMed Central

    Di Berardino, M; Dimroth, P

    1996-01-01

    We report here a new mode of coupling between the chemical and vectorial reaction explored for the oxaloacetate decarboxylase Na+ pump from Klebsiella pneumoniae. The membrane-bound beta-subunit is responsible for the decarboxylation of carboxybiotin and the coupled translocation of Na+ ions across the membrane. The biotin prosthetic group which is attached to the alpha-subunit becomes carboxylated by carboxyltransfer from oxaloacetate. The two conserved aspartic acid residues within putative membrane-spanning domains of the beta-subunit (Asp149 and Asp203) were exchanged by site-directed mutagenesis. Mutants D149Q and D149E retained oxaloacetate decarboxylase and Na+ transport activities. Mutants D203N and D203E, however, had lost these two activities, but retained the ability to form the carboxybiotin enzyme. Direct participation of Asp203 in the catalysis of the decarboxylation reaction is therefore indicated. In addition, all previous and present data on the enzyme support a model in which the same aspartic acid residue provides a binding site for the metal ion catalysing its movement across the membrane. The model predicts that asp203 in its dissociated form binds Na+ and promotes its translocation, while the protonated residue transfers the proton to the acid-labile carboxybiotin which initiates its decarboxylation. Strong support for the model comes from the observation that Na+ transport by oxaloacetate decarboxylation is accompanied by H+ transport in the opposite direction. The inhibition of oxaloacetate decarboxylation by high Na+ concentrations in a pH-dependent manner is also in agreement with the model. Images PMID:8617230

  19. A Flash-ADC data acquisition system developed for a drift chamber array and a digital filter algorithm for signal processing

    NASA Astrophysics Data System (ADS)

    Yi, Han; Lü, Li-Ming; Zhang, Zhao; Cheng, Wen-Jing; Ji, Wei; Huang, Yan; Zhang, Yan; Li, Hong-Jie; Cui, Yin-Ping; Lin, Ming; Wang, Yi-Jie; Duan, Li-Min; Hu, Rong-Jiang; Xiao, Zhi-Gang

    2016-11-01

    A Flash-ADC data acquisition (DAQ) system has been developed for the drift chamber array designed for the External-Target-Experiment at the Cooling Storage Ring at the Heavy Ion Research Facility, Lanzhou. The simplified readout electronics system has been developed using the Flash-ADC modules and the whole waveform in the sampling window is obtained, with which the time and energy information can be deduced with an offline processing. A digital filter algorithm has been developed to discriminate the noise and the useful signal. With the digital filtering process, the signal to noise ratio (SNR) is increased and a better time and energy resolution can be obtained. Supported by National Basic Research Program of China (973) (2015CB856903 and 2014CB845405), partly by National Science Foundation of China (U1332207 and 11375094), and by Tsinghua University Initiative Scientific Research Program

  20. Experimental demonstration of a real-time high-throughput digital DC blocker for compensating ADC imperfections in optical fast-OFDM receivers.

    PubMed

    Zhang, Lu; Ouyang, Xing; Shao, Xiaopeng; Zhao, Jian

    2016-06-27

    Performance degradation induced by the DC components at the output of real-time analogue-to-digital converter (ADC) is experimentally investigated for optical fast-OFDM receiver. To compensate this degradation, register transfer level (RTL) circuits for real-time digital DC blocker with 20GS/s throughput are proposed and implemented in field programmable gate array (FPGA). The performance of the proposed real-time digital DC blocker is experimentally investigated in a 15Gb/s optical fast-OFDM system with intensity modulation and direct detection over 40 km standard single-mode fibre. The results show that the fixed-point DC blocker has negligible performance penalty compared to the offline floating point one, and can overcome the error floor of the fast OFDM receiver caused by the DC components from the real-time ADC output.