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Sample records for lyase pal activity

  1. Phenylalanine ammonia-lyase (PAL) gene activity in response to ...

    African Journals Online (AJOL)

    Phenylalanine ammonia-lyase (PAL) catalyzes the biosynthesis of rosmarinic acid (RA), tyrosine and phenylalanine are the precursors of RA, while proline drives metabolite precursors toward Shikimate and phenylpropanoid pathway ending with the production of RA. The aim of this study was to investigate the PAL gene ...

  2. Suppressed phenylalanine ammonia-lyase activity after heat shock in transgenic Nicotiana plumbaginifolia containing an Arabidopsis HSP18.2-parsley PAL2 chimera gene.

    Science.gov (United States)

    Moriwaki, M; Yamakawa, T; Washino, T; Kodama, T; Igarashi, Y

    1999-01-01

    The activity of phenylalanine ammonia-lyase (PAL; EC 4.3.1.5) after heat shock (HS) in leaves and buds of transgenic Nicotiana plumbaginifolia containing an Arabidopsis HSP18.2 promoter-parsley phenylalanine ammonia-lyase 2 (HSP18.2-PAL2) chimera gene was examined. Immediately after HS treatment at 44 degrees C for 5 h, the PAL activity in both transgenic and normal (untransformed) plants was 35-38% lower than that before HS. At normal temperature (25-26 degrees C), the PAL activity recovered within 5 h of ending the HS treatment in normal plants, but not until 12-24 h in transgenic plants containing the HSP18.2-PAL2 gene. Reverse transcription-polymerase chain reaction (RT-PCR) analysis revealed the presence of parsley PAL2 mRNA in transgenic plants, which remained for 8-12 h following 5-h HS at 44 degrees C; the mRNA was not observed before HS. The content of chlorogenic acid (CGA; 3-caffeoylquinic acid) decreased drastically 8-12 h after HS in transgenic plants, but only slightly in normal plants. Thus, the decrease in PAL activity accompanied by expression of the parsley PAL2 gene after HS treatment corresponded to the decrease in CGA synthesis. These results might be attributed to post-transcriptional degradation of endogenous PAL mRNA triggered by transcription of the transgene.

  3. Phenylalanine ammonia-lyase (pal) and peroxidase activity in brown rust infected tissues of pakistani wheat cultivars

    International Nuclear Information System (INIS)

    Riaz, A.; Tahir, M.I.

    2014-01-01

    Besides other factors resistance and susceptibility is the outcome of biochemical processes such as activities of defense-related enzymes. So in this study, Phenylalanine ammonia-lyase (PAL) and Peroxidase activity of resistant (Inqilab-91) and susceptible (Kirin-95) wheat cultivars were determined through spectrophotometer to address the biochemical aspect related to the disease after 8 hours, 24 hours, 48 hours and 72 hours of leaf rust inoculation. The results have shown that these enzymes were present in both the resistant and susceptible cultivars but the activity was more pronounced in the resistant one. The effect of PAL and peroxidase activity was also investigated among inoculated and uninoculated plants within the same cultivar. The activity of both PAL and peroxidase were more significant in inoculated ones. The results have shown that the after 72 hours of inoculation Inqilab-91 had more PAL activity i.e., 5.47 IU/ml/min than in Kirin-95 i.e., 2.08 IU/ml/min at 270 nm. While peroxidase activity in Inqilab-91 was 6.41 IU/ml/min and in Kirin-95, 3.66 IU/ml/min after 72 hours of inoculation, observed under 470 nm wavelength. Increase in one's activity increases the other enzyme's activity. The activity was more prominent after 72 hours of infection as pathogen had successfully established itself in the host plant tissue. The activities of these enzymes act as plants active defense mechanism against the attack of pathogen. (author)

  4. Reduced phenylalanine ammonia-lyase and tyrosine ammonia-lyase activities and lignin synthesis in wheat grown under low pressure sodium lamps

    Science.gov (United States)

    Guerra, D.; Anderson, A. J.; Salisbury, F. B.

    1985-01-01

    Wheat (Triticum aestivum L. cv Fremont) grown in hydroponic culture under 24-hour continuous irradiation at 560 to 580 micromoles per square meter per second from either metalhalide (MH), high pressure sodium (HPS), or low pressure sodium (LPS) lamps reached maturity in 70 days. Grain yields were similar under all three lamps, although LPS-grown plants lodged at maturity. Phenylalanine ammonia-lyase (PAL) and a tyrosine ammonia lyase (TAL) with lesser activity were detected in all extracts of leaf, inflorescence, and stem. Ammonia-lyase activities increased with age of the plant, and plants grown under the LPS lamp displayed PAL and TAL activities lower than wheat cultured under MH and HPS radiation. Greenhouse solar-grown wheat had the highest PAL and TAL activities. Lignin content of LPS-grown wheat was also significantly reduced from that of plants grown under MH or HPS lamps or in the greenhouse, showing a correlation with the reduced PAL and TAL activities. Ratios of far red-absorbing phytochrome to total phytochrome were similar for all three lamps, but the data do not yet warrant a conclusion about specific wavelengths missing from the LPS lamps that might have induced PAL and TAL activities in plants under the other lamps.

  5. Regulation of a phenylalanine ammonia lyase (BbPAL) by calmodulin in response to environmental changes in the entomopathogenic fungus Beauveria bassiana.

    Science.gov (United States)

    Kim, Jiyoung; Park, Hyesung; Han, Jae-Gu; Oh, Junsang; Choi, Hyung-Kyoon; Kim, Seong Hwan; Sung, Gi-Ho

    2015-11-01

    Phenylalanine ammonia lyase (PAL, E.C. 4.3.1.5) catalyses the deamination of L -phenylalanine to trans-cinnamic acid and ammonia, facilitating a critical step in the phenylpropanoid pathway that produces a variety of secondary metabolites. In this study, we isolated BbPAL gene in the entomopathogenic fungus Beauveria bassiana. According to multiple sequence alignment, homology modelling and in vitro PAL activity, we demonstrated that BbPAL acts as a typical PAL enzyme in B. bassiana. BbPAL interacted with calmodulin (CaM) in vitro and in vivo, indicating that BbPAL is a novel CaM-binding protein. The functional role of CaM in BbPAL action was to negatively regulate the BbPAL activity in B. bassiana. High-performance liquid chromatography analysis revealed that L -phenylalanine was reduced and trans-cinnamic acid was increased in response to the CaM inhibitor W-7. Dark conditions suppressed BbPAL activity in B. bassiana, compared with light. In addition, heat and cold stresses inhibited BbPAL activity in B. bassiana. Interestingly, these negative effects of BbPAL activity by dark, heat and cold conditions were recovered by W-7 treatment, suggesting that the inhibitory mechanism is mediated through stimulation of CaM activity. Therefore, this work suggests that BbPAL plays a role in the phenylpropanoid pathway mediated by environmental stimuli via the CaM signalling pathway. © 2015 Society for Applied Microbiology and John Wiley & Sons Ltd.

  6. Isolation and Functional Characterization of a Phenylalanine Ammonia-Lyase Gene (SsPAL1 from Coleus (Solenostemon scutellarioides (L. Codd

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    Qinlong Zhu

    2015-09-01

    Full Text Available Phenylalanine ammonia-lyase (PAL is the first enzyme involved in the phenylpropanoid pathway and plays important roles in the secondary metabolisms, development and defense of plants. To study the molecular function of PAL in anthocyanin synthesis of Coleus (Solenostemon scutellarioides (L. Codd, a Coleus PAL gene designated as SsPAL1 was cloned and characterized using a degenerate oligonucleotide primer PCR and RACE method. The full-length SsPAL1 was 2450 bp in size and consisted of one intron and two exons encoding a polypeptide of 711 amino acids. The deduced SsPAL1 protein showed high identities and structural similarities with other functional plant PAL proteins. A series of putative cis-acting elements involved in transcriptional regulation, light and stress responsiveness were found in the upstream regulatory sequence of SsPAL1. Transcription pattern analysis indicated that SsPAL1 was constitutively expressed in all tissues examined and was enhanced by light and different abiotic factors. The recombinant SsPAL1 protein exhibited high PAL activity, at optimal conditions of 60 °C and pH 8.2. Although the levels of total PAL activity and total anthocyanin concentration have a similar variation trend in different Coleus cultivars, there was no significant correlation between them (r = 0.7529, p > 0.1, suggesting that PAL was not the rate-limiting enzyme for the downstream anthocyanin biosynthetic branch in Coleus. This study enables us to further understand the role of SsPAL1 in the phenylpropanoid (flavonoids, anthocyanins biosynthesis in Coleus at the molecular level.

  7. Molecular and analysis of a phenylalanine ammonia-lyase gene (LrPAL2) from Lycoris radiata.

    Science.gov (United States)

    Jiang, Yumei; Xia, Bing; Liang, Lijian; Li, Xiaodan; Xu, Sheng; Peng, Feng; Wang, Ren

    2013-03-01

    Phenylalanine ammonia-lyase (PAL), the first enzyme of phenylpropanoid biosynthesis, participates in the biosynthesis of flavonoids, lignins, stilbenes and many other compounds. In this study, we cloned a 2,326 bp full-length PAL2 gene from Lycoris radiata by using degenerate oligonucleotide primer PCR (DOP-PCR) and the rapid amplification of cDNA ends method. The cDNA contains a 2,124 bp coding region encoding 707 amino acids. The LrPAL2 shares about 77.0 % nucleic acid identity and 83 % amino acid identity with LrPAL1. Furthermore, genome sequence analysis demonstrated that LrPAL2 gene contains one intron and two exons. The 5' flanking sequence of LrPAL2 was also cloned by self-formed adaptor PCR (SEFA-PCR), and a group of putative cis-acting elements such as TATA box, CAAT box, G box, TC-rich repeats, CGTCA motif and TCA-element were identified. The LrPAL2 was detected in all tissues examined, with high abundance in bulbs at leaf sprouting stage and in petals at blooming stage. Besides, LrPAL2 drastically responded to MJ, SNP and UV, moderately responded to GA and SA, and a little increased under wounding. Comparison of LrPAL2 expression and LrPAL1 expression demonstrated that LrPAL2 can be more significantly induced than LrPAL1 under the above treatments, and LrPAL2 transcripts accumulated prominently at blooming stage, especially in petals, while LrPAL1 transcripts did not accumulated significantly at blooming stage. All these results suggested that LrPAL2 might play distinct roles in different branches of the phenylpropanoid pathway.

  8. Phenylalanine ammonia lyase from Arabidopsis thaliana (AtPAL2): A potent MIO-enzyme for the synthesis of non-canonical aromatic alpha-amino acids: Part I: Comparative characterization to the enzymes from Petroselinum crispum (PcPAL1) and Rhodosporidium toruloides (RtPAL).

    Science.gov (United States)

    Dreßen, Alana; Hilberath, Thomas; Mackfeld, Ursula; Billmeier, Arne; Rudat, Jens; Pohl, Martina

    2017-09-20

    Phenylalanine ammonia lyase (PAL) from Arabidopsis thaliana (AtPAL2) was comparatively characterized to the well-studied enzyme from parsley (PcPAL1) and Rhodosporidium toruloides (RtPAL) with respect to kinetic parameters for the deamination and the amination reaction, pH- and temperature optima and the substrate range of the amination reaction. Whereas both plant enzymes are specific for phenylalanine, the bifunctional enzyme from Rhodosporidium toruloides shows K M -values for L-Phe and L-Tyr in the same order of magnitude and, compared to both plant enzymes, a 10-15-fold higher activity. At 30°C all enzymes were sufficiently stable with half-lives of 3.4days (PcPAL1), 4.6days (AtPAL2) and 9.7days (RtPAL/TAL). Very good results for the amination of various trans-cinnamic acid derivatives were obtained using E. coli cells as whole cell biocatalysts in ammonium carbonate buffer. Investigation of the substrate ranges gave interesting results for the newly tested enzymes from A. thaliana and R. toruloides. Only the latter accepts besides 4-hydroxy-CA also 3-methoxy-4-hydroxy-CA as a substrate, which is an interesting intermediate for the formation of pharmaceutically relevant L-Dopa. AtPAL2 is a very good catalyst for the formation of (S)-3-F-Phe, (S)-4-F-Phe and (S)-2-Cl-Phe. Such non-canonical amino acids are valuable building blocks for the formation of various drug molecules. Copyright © 2017 Elsevier B.V. All rights reserved.

  9. The effect of cadmium on phenylalanine ammonia lyase activity and lipid peroxidation in pepper (Capsicum annuum L. seedlings

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    Esra Koç

    2015-04-01

    Full Text Available In this study, the effect on differrent concentrations (20, 40, 80µM ve 100 µM CdCl2 of cadmium (CdCl2 on the activity of phenylalanine ammonia-lyase (PAL and lipid peroxidation amount in leaf and stem of Kahramanmaraş- Hot (Capsicum annum L. pepper seedlings were researched. Activity of phenylalanine ammonia-lyase (PAL, the first enzyme in the phenylpropanoid biosynthetic pathway, was increased at 2 and 4 days in KM-Hot plants exposed to CdCl2 stress. The highest PAL activity was detected in 20 μM CdCl2 application, on the four day after the application in the leaves of KM-Hot pepper. Moreover, it was observed that treatment of pepper with Cd led to an increased the rate of lipid peroxidation (which is indicated by increasing MDA content in the leaf and stem tissues. The highest MDA content was detected in 80 μM CdCl2 application, on the four day after the application in the leaf tissues. These results suggest that the activation of PAL may be associated with increased production of MDA

  10. Expression and enzymatic activity of phenylalanine ammonia-lyase and p-coumarate 3-hydroxylase in mango (Mangifera indica 'Ataulfo') during ripening.

    Science.gov (United States)

    Palafox-Carlos, H; Contreras-Vergara, C A; Muhlia-Almazán, A; Islas-Osuna, M A; González-Aguilar, G A

    2014-05-16

    Phenylalanine ammonia lyase (PAL) and p-coumarate 3-hydroxylase (C3H) are key enzymes in the phenylpropanoid pathway. The relative expression of PAL and C3H was evaluated in mango fruit cultivar 'Ataulfo' in four ripening stages (RS1, RS2, RS3, and RS4) by quantitative polymerase chain reaction. In addition, enzyme activity of PAL and C3H was determined in mango fruits during ripening. The PAL levels were downregulated at the RS2 and RS3 stages, while C3H levels were upregulated in fruits only at RS3. The enzyme activity of PAL followed a pattern that was different from that of the PAL expression, thus suggesting regulation at several levels. For C3H, a regulation at the transcriptional level is suggested because a similar pattern was revealed by its activity and transcript level. In this study, the complexity of secondary metabolite biosynthesis regulation is emphasized because PAL and C3H enzymes are involved in the biosynthesis of several secondary metabolites that are active during all mango ripening stages.

  11. Optimization of oligomeric enzyme activity in ionic liquids using Rhodotorula glutinis yeast phenylalanine ammonia lyase.

    Science.gov (United States)

    Barron, Christiaan C; Sponagle, Brandon J D; Arivalagan, Pugazhendhi; D'Cunha, Godwin B

    2017-01-01

    Phenylalanine ammonia lyase (E.C.4.3.1.24, PAL) activity of Rhodotorula glutinis yeast has been demonstrated in four commonly used ionic liquids. PAL forward reaction was carried out in 1-butyl-3-methylimidazolium methyl sulfate ([BMIM][MeSO 4 ]), 1-butyl-3-methylimidazolium tetrafluoroborate ([BMIM][BF 4 ]), 1-butyl-3-methylimidazolium hexafluorophosphate ([BMIM][PF 6 ]) and 1-butyl-3-methylimidazolium lactate ([BMIM][lactate]). Our experiments have revealed that PAL is catalytically active in ionic liquids and the enzyme activity in ([BMIM][PF 6 ]) is comparable to that obtained in aqueous buffer medium. Different conditions were optimized for maximal PAL forward activity including time of incubation (30.0min) L -phenylalanine substrate concentration (30.0mM), nature of buffer (50.0mM Tris-HCl), pH (9.0), temperature (37°C), and speed of agitation (100 rev min -1 ). Under these optimized conditions, about 83% conversion of substrate to product was obtained for the PAL forward reaction that was determined using UV spectroscopy at 290nm. PAL reverse reaction in ([BMIM][PF 6 ]) was determined spectrophotometrically at 520nm; and about 59% substrate conversion was obtained. This data provides further knowledge in enzyme biocatalysis in non-aqueous media, and may be of importance when studying the function of other oligomeric/multimeric proteins and enzymes in ionic liquids. Copyright © 2016 Elsevier Inc. All rights reserved.

  12. Possible regulatory role of phenylalanine ammonia-lyase in the production of anthocyanins in asparagus (Asparagus officinalis L)

    NARCIS (Netherlands)

    Flores, F.B.; Oosterhaven, J.; Martinez-Madrid, M.C.; Romojaro, F.

    2005-01-01

    The regulatory role of phenylalanine ammonia-lyase (PAL) in the light-induced accumulation of anthocyanins in the epidermis of asparagus spears has been analysed. A correlation between the stimulation of PAL activity and the rise in total anthocyanin content has been observed. Light radiation

  13. [The relationship between activity and gene expression of phenylalanine ammonia-lyase and peel pitting in 'Fengjie' navel orange fruits].

    Science.gov (United States)

    Li, Zheng-Guo; Gao, Xue; Fan, Jing; Yang, Ying-Wu; Li, Dao-Gao; Kanellis, Angelos K

    2006-06-01

    Citrus fruit is prone to develop peel pitting during development and storage, which greatly decreases its fresh market value because of the deterioration of the peel. In the present study, we have examined the effect of different temperatures (15 degrees C and 4 degrees C), waxing and mechanical damage on the changes in the activity of phenylalanine ammonia-lyase (PAL) and the incidence of peel pitting in 'Fengjie' navel orange (Citrus sinensis Osbeck) fruits. The expression levels of PAL2, PAL6 genes in the peel during the development of peel pitting have been investigated through semi-quantitative PCR method. The incidence of peel pitting was greatly enhanced by waxing and mechanical damage and was decreased in lower temperature storage (4 degrees C) (Fig.1). Waxing and mechanical damage might be the important factors inducing peel pitting and suitable low temperature could decrease the incidence of this disease. The PAL activity increased during the whole storage period in accordance with the development of this pitting (Fig.2). The expression levels of PAL2 and PAL6 genes in damaged peel were higher than those in healthy peel and the expression of PAL2 is much more higher than that of PAL6 (Figs.4 and 5). The results suggested that the enzyme activity of PAL, along with the expression of PAL2 gene is highly related to this peel pitting occurred on 'Fengjie' navel orange fruits.

  14. Molecular Cloning and Sequence Analysis of a Phenylalanine Ammonia-Lyase Gene from Dendrobium

    Science.gov (United States)

    Cai, Yongping; Lin, Yi

    2013-01-01

    In this study, a phenylalanine ammonia-lyase (PAL) gene was cloned from Dendrobium candidum using homology cloning and RACE. The full-length sequence and catalytic active sites that appear in PAL proteins of Arabidopsis thaliana and Nicotiana tabacum are also found: PAL cDNA of D. candidum (designated Dc-PAL1, GenBank No. JQ765748) has 2,458 bps and contains a complete open reading frame (ORF) of 2,142 bps, which encodes 713 amino acid residues. The amino acid sequence of DcPAL1 has more than 80% sequence identity with the PAL genes of other plants, as indicated by multiple alignments. The dominant sites and catalytic active sites, which are similar to that showing in PAL proteins of Arabidopsis thaliana and Nicotiana tabacum, are also found in DcPAL1. Phylogenetic tree analysis revealed that DcPAL is more closely related to PALs from orchidaceae plants than to those of other plants. The differential expression patterns of PAL in protocorm-like body, leaf, stem, and root, suggest that the PAL gene performs multiple physiological functions in Dendrobium candidum. PMID:23638048

  15. Characterization of promoter of EgPAL1, a novel PAL gene from the oil palm Elaeis guineensis Jacq.

    Science.gov (United States)

    Yusuf, Chong Yu Lok; Abdullah, Janna Ong; Shaharuddin, Noor Azmi; Abu Seman, Idris; Abdullah, Mohd Puad

    2018-02-01

    The oil palm EgPAL1 gene promoter and its regulatory region were functional as a promoter in the heterologous system of Arabidopsis according to the cis-acting elements present in that region. The promoter was developmentally regulated, vascular tissue specific and responsive to water stress agents. Phenylalanine ammonia lyase (PAL, EC 4.3.1.24) is the key enzyme of the phenylpropanoid pathway which plays important roles in plant development and adaptation. To date, there is no report on the study of PAL from oil palm (Elaeis guineensis), an economically important oil crop. In this study, the 5' regulatory sequence of a highly divergent oil palm PAL gene (EgPAL1) was isolated and fused with GUS in Arabidopsis to create two transgenic plants carrying the minimal promoter with (2302 bp) and without its regulatory elements (139 bp). The regulatory sequence contained cis-acting elements known to be important for plant development and stress response including the AC-II element for lignin biosynthesis and several stress responsive elements. The promoter and its regulatory region were fully functional in Arabidopsis. Its activities were characterised by two common fundamental features of PAL which are responsive to plant internal developmental programme and external factors. The promoter was developmentally regulated in certain organs; highly active in young organs but less active or inactive in mature organs. The presence of the AC elements and global activity of the EgPAL1 promoter in all organs resembled the property of lignin-related genes. The existence of the MBS element and enhancement of the promoter activity by PEG reflected the behaviour of drought-responsive genes. Our findings provide a platform for evaluating oil palm gene promoters in the heterologous system of Arabidopsis and give insights into the activities of EgPAL1 promoter in oil palm.

  16. Expression and properties of the highly alkalophilic phenylalanine ammonia-lyase of thermophilic Rubrobacter xylanophilus.

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    Klaudia Kovács

    Full Text Available The sequence of a phenylalanine ammonia-lyase (PAL; EC: 4.3.1.24 of the thermophilic and radiotolerant bacterium Rubrobacter xylanophilus (RxPAL was identified by screening the genomes of bacteria for members of the phenylalanine ammonia-lyase family. A synthetic gene encoding the RxPAL protein was cloned and overexpressed in Escherichia coli TOP 10 in a soluble form with an N-terminal His6-tag and the recombinant RxPAL protein was purified by Ni-NTA affinity chromatography. The activity assay of RxPAL with l-phenylalanine at various pH values exhibited a local maximum at pH 8.5 and a global maximum at pH 11.5. Circular dichroism (CD studies showed that RxPAL is associated with an extensive α-helical character (far UV CD and two distinctive near-UV CD peaks. These structural characteristics were well preserved up to pH 11.0. The extremely high pH optimum of RxPAL can be rationalized by a three-dimensional homology model indicating possible disulfide bridges, extensive salt-bridge formation and an excess of negative electrostatic potential on the surface. Due to these properties, RxPAL may be a candidate as biocatalyst in synthetic biotransformations leading to unnatural l- or d-amino acids or as therapeutic enzyme in treatment of phenylketonuria or leukemia.

  17. Cloning, expression and characterization of phenylalanine ammonia-lyase from Rhodotorula glutinis.

    Science.gov (United States)

    Zhu, Longbao; Cui, Wenjing; Fang, Yueqin; Liu, Yi; Gao, Xinxing; Zhou, Zhemin

    2013-05-01

    The industrial-scale production of phenylalanine ammonia-lyase (PAL) mainly uses strains of Rhodotorula. However, the PAL gene from Rhodotorula has not been cloned. Here, the full-length gene of PAL from Rhodotorula glutinis was isolated. It was 2,121 bp, encoding a polypeptide with 706 amino acids and a calculated MW of 75.5 kDa. Though R. glutinis is an anamorph of Rhodosporium toruloides, the amino acid sequences of PALs them are not the same (about 74 % identity). PAL was expressed in E. coli and characterized. Its specific activity was 4.2 U mg(-1) and the k cat/K m was 1.9 × 10(4) mM(-1) s(-1), exhibiting the highest catalytic ability among the reported PALs. The genetic and biochemical information reported here should facilitate future application in industry.

  18. Molecular cloning and sequence analysis of a phenylalanine ammonia-lyase gene from dendrobium.

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    Qing Jin

    Full Text Available In this study, a phenylalanine ammonia-lyase (PAL gene was cloned from Dendrobium candidum using homology cloning and RACE. The full-length sequence and catalytic active sites that appear in PAL proteins of Arabidopsis thaliana and Nicotiana tabacum are also found: PAL cDNA of D. candidum (designated Dc-PAL1, GenBank No. JQ765748 has 2,458 bps and contains a complete open reading frame (ORF of 2,142 bps, which encodes 713 amino acid residues. The amino acid sequence of DcPAL1 has more than 80% sequence identity with the PAL genes of other plants, as indicated by multiple alignments. The dominant sites and catalytic active sites, which are similar to that showing in PAL proteins of Arabidopsis thaliana and Nicotiana tabacum, are also found in DcPAL1. Phylogenetic tree analysis revealed that DcPAL is more closely related to PALs from orchidaceae plants than to those of other plants. The differential expression patterns of PAL in protocorm-like body, leaf, stem, and root, suggest that the PAL gene performs multiple physiological functions in Dendrobium candidum.

  19. Stabilization of Phenylalanine Ammonia Lyase from Rhodotorula glutinis by Encapsulation in Polyethyleneimine-Mediated Biomimetic Silica.

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    Cui, Jiandong; Liang, Longhao; Han, Cong; Lin Liu, Rong

    2015-06-01

    Phenylalanine ammonia lyase (PAL) from Rhodotorula glutinis was encapsulated within polyethyleneimine-mediated biomimetic silica. The main factors in the preparation of biomimetic silica were optimized by response surface methodology (RSM). Compared to free PAL (about 2 U), the encapsulated PAL retained more than 43 % of their initial activity after 1 h of incubation time at 60 °C, whereas free PAL lost most of activity in the same conditions. It was clearly indicated that the thermal stability of PAL was improved by encapsulation. Moreover, the encapsulated PAL exhibited the excellent stability of the enzyme against denaturants and storage stability, and pH stability was improved by encapsulation. Operational stability of 7 reaction cycles showed that the encapsulated PAL was stable. Nevertheless, the K m value of encapsulated PAL in biomimetic silica was higher than that of the free PAL due to lower total surface area and increased mass transfer resistance.

  20. Cadmium-induced changes in pigments, total phenolics, and phenylalanine ammonia-lyase activity in fronds of Azolla imbricata.

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    Dai, Ling-Peng; Xiong, Zhi-Ting; Huang, Yu; Li, Min-Jing

    2006-10-01

    This study was designed to examine the effects of cadmium on several color-related parameters (including chlorophyll, carotenoid, and anthocyanin), total phenolics, and phenylalanine ammonia-lyase (PAL) activity in an aquatic fern species Azolla imbricate (A. imbricata). Cd accumulation and effects in the fronds were closely related with Cd concentration in the growth medium. The fronds under 0.5 mg/L Cd treatment turned red on the 3rd day, and this color change also appeared under 0.05 and 0.1 mg/L Cd treatment on the 5th day. Correlated with the color change, the contents of chlorophyll and carotenoid in the fronds significantly decreased in the presence of high Cd concentrations, while the anthocyanin content increased during the experiment. Significant increase in total phenolics content and PAL activity were also detected during Cd treatment. The results suggested that the Cd-induced change in color of fronds might be due to the decrease in chlorophyll and carotenoid and the increase in anthocyanin. Anthocyanin, total phenolics and their biosynthesis-related PAL might play a role in detoxification of Cd in A. imbricata.

  1. Developmental role of phenylalanine-ammonia-lyase (PAL) and cinnamate 4-hydroxylase (C4H) genes during adventitious rooting of Juglans regia L. microshoots.

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    Cheniany, Monireh; Ganjeali, Ali

    2016-12-01

    Phenylalanine-ammonia-lyase and cinnamate-4-hydroxylase play important role in the phenylpropanoid pathway, which produces many biologically important secondary metabolites participating in normal plant development. Flavonol quercetin is the main representant of these compounds that has been identified in numerous Juglans spp. In this survey, the developmental expression patterns of PAL and C4H genes during in vitro rooting of two walnut cultivars 'Sunland' and 'Howard' was examined by RT-PCR. To understand the potential role in rooting, the changing pattern of endogenous content of quercetin was also analyzed by HPLC. The 'Sunland' with better capacity to root had more quercetin content during the "inductive phase" of rooting than 'Howard'. In each cultivar, the level of PAL transcripts showed the same behavior with the changing patterns of quercetin during root formation of microshoots. The positive correlation between the changes of quercetin and PAL-mRNA indicated that PAL gene may have an immediate effect on flavonoid pathway metabolites including quercetin. Although the behavioral change of C4H expression was similar in both cultivars during root formation (with significantly more level for 'Howard'), it was not coincide with the changes of quercerin concentrations. Our results showed that C4H function is important for the normal development, but its transcriptional regulation does not correlate with quercetin as an efficient phenolic compound for walnut rhizogenesis.

  2. Evaluation of PAL activity, Phenolic and Flavonoid Contents in Three Pistachio (Pistacia vera L. Cultivars Grafted onto Three Different Rootstocks

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    N. Nadernejad

    2013-08-01

    Full Text Available Phenylalanine ammonia lyase (PAL is a biochemical marker of the environmental stress and plays a pivotal role in phenolic synthesis. The lower ROS level and oxidative damage was observed in grafted plants and the rootstocks have a profound influence on the biochemical composition, especially phenolic compounds. Regarding the importance of the effect rootstocks have on scion in pistachio trees, this study was carried out to assess and compare three pistachio cultivars ("Ahmadaghaii", "Ohadi" and "Kallehghuchi" on three rootstocks (Mutica, Ahli, Sarakhs. PAL activity, phenolic compounds, flavonoid and anthocyanin contents in leaves, flowers and fruits were measured toward the selection of the most suitable and compatible rootstock/scion resistant to environmental stresses. The results showed that PAL activity was different among the cultivars and organs. A positive correlation was observed between PAL activity and phenolic compounds in the leaves and flowers of Mutica- Ahmadaghaii, suggesting that it is more resistant than the others to environmental stresses. PAL activity and total phenolics in fruits of pistachio suffered a decrease when the maturation processes began. The hulls of the pistachio fruits contained high levels of phenolic compounds especially in Mutica-Ahmadaghaii suggesting its function as a protective layer and a defense chemical against ultraviolet radiation and pathogen. Our results indicated the presence of a number of bioactive compounds in kernels with the highest amount belonging to Mutica- Ahmadaghaii, and therefore it is concluded that pistachio rootstocks may affect the antioxidant compounds in kernels.

  3. Cooperative functioning between phenylalanine ammonia lyase and isochorishmate synthase activities contributes to salicylic acid biosynthesis in soybean

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    Salicylic acid (SA), an essential regulator of plant defense, is derived from chorismate via either the phenylalanine ammonia lyase (PAL), or the isochorishmate synthase (ICS) catalyzed steps. The ICS pathway is thought to be the primary contributor of defense-related SA, at least in Arabidopsis. We...

  4. A phenylalanine ammonia-lyase ortholog (PkPAL1) from Picrorhiza kurrooa Royle ex. Benth: molecular cloning, promoter analysis and response to biotic and abiotic elicitors.

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    Bhat, Wajid Waheed; Razdan, Sumeer; Rana, Satiander; Dhar, Niha; Wani, Tariq Ahmad; Qazi, Parvaiz; Vishwakarma, Ram; Lattoo, Surrinder K

    2014-09-01

    Picrorhiza kurrooa Royle ex Benth. is a highly reputed medicinal herb utilised in the preparation of a number of herbal drug formulations, principally due to the presence of novel monoterpene iridoid glycosides kenned as picrosides. Phenylalanine ammonia-lyase catalyses an important rate-limiting step in phenylpropanoid pathway and supplies precursors like cinnamic acid, vanillic acid, ferulic acid, etc., to a variety of secondary metabolites including picrosides. The imperilled status of P. kurrooa coupled with lack of information regarding biogenesis of picrosides necessitates deciphering the biosynthetic pathway for picrosides. In the present study, a PAL gene, designated PkPAL1 was isolated from P. kurrooa. The cDNA is 2312 bp in length, consisting of an ORF of 2142 bp encoding for a 713 amino acid protein having a predicted molecular weight of 77.66 kDa and an isoelectric point of pH 6.82. qRT-PCR analysis of various tissues of P. kurrooa showed that PkPAL1 transcript levels were highest in the leaves, consistent with picroside accumulation pattern. Using Genome walking, a 718 bp promoter region was also isolated resulting in identification of distinct cis-regulatory elements including TGA-element, TGACG-motif, CGTCA-motif, etc. qRT-PCR indicated up-regulation of PkPAL1 by methyl jasmonate, salicylic acid, 2,4-dicholorophenoxy acetic acid and UV-B elicitations that corroborated positively with the identified cis-elements within the promoter region. Moreover, altitude was found to have a positive effect on the PkPAL1 transcript levels, driving the expression of PkPAL1 abundantly. Based on docking analysis, we identified eight residues as potentially essential for substrate binding in PkPAL1. Copyright © 2014 Elsevier B.V. All rights reserved.

  5. Control of phenylalanine ammonia-lyase gene promoters from pea by UV radiation

    International Nuclear Information System (INIS)

    Pluskota, W.E.; Michalczyk, D.J.; Gorecki, R.J.

    2005-01-01

    The gene fusion system was used to study UV light-control of PS PAL1 and PS PAL2 genes encoding phenylalanine ammonia-lyase of pea. The induction of pea PAL promoters was analysed in transgenic tobacco plants. Binary plasmids (derivatives of pBI101.2 vector) containing 5' regulatory fragments of PS PAL1 and PS PAL2 linked to reporter genes (GUS, LUC) were constructed. The analyses were performed with the use of single constructs (containing one variant of PS PAL promoter and one reporter gene) and dual constructs (containing both PS PAL1 and PS PAL2 promoters connected with different reporter genes). The use of dual constructs enabled the evaluation of both PS PAL promoters activity in the same plant. The analyses of in vitro grown plants have shown that both PAL promoters are strongly induced in leaves subjected to UV radiation. In some cases, the UV-stimulated expression exceeded the exposed areas. This phenomenon was observed more often in the leaves of plants containing the PS PAL1::GUS than PS PAL2::GUS construct. Removal of boxes 2, 4, 5 from PS PAL1 promoter and deletion of its 5' end region (-339 to -1394) decreases the level of gene expression but does not eliminate its responsiveness to UV

  6. Phenylalanine ammonia-lyase through evolution: A bioinformatic approach

    Directory of Open Access Journals (Sweden)

    Shiva Hemmati

    2015-03-01

    Full Text Available Phenylalanine ammonia-lyase (PAL is the first entry enzyme of the phenylpropanoid pathway that converts phenylalanine to cinnamic acid which is the precursor of various secondary metabolites. PAL is recently formulated for phenylketonuric patients in pegylated forms; therefore, screening a PAL with the highest affinity to the substrate is of a great importance. PAL exists in all higher plants and some fungi and few bacteria. Ancestors of land plants have been adopted by evolving metabolic pathways. A multi-gene family encodes PAL by gene duplication events in most plants. In this study, the taxonomic distribution and phylogeny of pal gene found in land plants, fungi and bacteria have been analyzed. It seems that the ancestor of plants acquired a pal gene via horizontal gene transfer in symbioses with bacteria and fungi. Gymnosperms have kept a diverse set of pal genes that arose from gene duplication events. In angiosperms, after the divergence of dicotyledons from monocots, pal genes were duplicated many times. The close paralogues of pal genes in some species indicate expansion of gene families after the divergence in plant pal gene evolution. Interestingly, some of the plant pals clustered by species in a way that pals within one species are more closely related to each other than to homologs in the other species which indicates this duplication event occurred more recently.

  7. Synthesis of specifically labelled L-phenylalanines using phenylalanine ammonia lyase activity

    International Nuclear Information System (INIS)

    Haedener, A.; Tamm, Ch.

    1987-01-01

    Specifically labelled L-phenylalanines have been prepared using a variety of classical synthetic methods in combination with phenylalanine ammonia lyase (PAL) enzyme activity of the yeast Rhodosporidium toruloides ATCC 10788 or Rhodotorula glutinis IFO 0559, respectively. Thus, L-[2- 2 H]phenyl-[2- 2 H]alanine was formed from (E) -[2,2'- 2 H 2 ]cinnamic acid and ammonia in 46% yield, whereas L-phenyl-[2- 13 C, 15 N]alanine was obtained from (E)-[2- 13 C]cinnamic acid in 45% overall yield. Generally, labelled cinnamic acids were recovered in pure form from the reaction mixture, with a loss of 6-8%. Likewise, unchanged 15 NH 3 was reisolated as 15 NH 4 Cl after steam distillation with overall losses of less than 4%. Labelled cinnamic acids were prepared by Knoevenagel condensations between appropriately labelled benzaldehydes and malonic acids. [2- 2 H]Benzaldehyde was obtained from 2-bromotoluene by decomposition of the corresponding Grignard reagent with 2 H 2 O and subsequent oxidation. Since simple molecules, most of them commercially available in labelled form or otherwise easily accessible, may serve as starting material, and due to its defined stereochemistry, the reaction catalysed by PAL opens a short and attractive route to specifically labelled L-phenylalanines. (author)

  8. Structural And Biochemical Characterization of the Therapeutic A. Variabilis Phenylalanine Ammonia Lyase

    Energy Technology Data Exchange (ETDEWEB)

    Wang, L.; Gamez, A.; Archer, H.; Abola, E.E.; Sarkissian, C.N.; Fitzpatrick, P.; Wendt, D.; Zhang, Y.; Vellard, M.; Bliesath, J.; Bell, S.; Lemont, J.; Scriver, C.R.; Stevens, R.C.

    2009-05-26

    We have recently observed promising success in a mouse model for treating the metabolic disorder phenylketonuria with phenylalanine ammonia lyase (PAL) from Rhodosporidium toruloides and Anabaena variabilis. Both molecules, however, required further optimization in order to overcome problems with protease susceptibility, thermal stability, and aggregation. Previously, we optimized PAL from R. toruloides, and in this case we reduced aggregation of the A. variabilis PAL by mutating two surface cysteine residues (C503 and C565) to serines. Additionally, we report the structural and biochemical characterization of the A. variabilis PAL C503S/C565S double mutant and carefully compare this molecule with the R. toruloides engineered PAL molecule. Unlike previously published PAL structures, significant electron density is observed for the two active-site loops in the A. variabilis C503S/C565S double mutant, yielding a complete view of the active site. Docking studies and N-hydroxysuccinimide-biotin binding studies support a proposed mechanism in which the amino group of the phenylalanine substrate is attacked directly by the 4-methylidene-imidazole-5-one prosthetic group. We propose a helix-to-loop conformational switch in the helices flanking the inner active-site loop that regulates accessibility of the active site. Differences in loop stability among PAL homologs may explain the observed variation in enzyme efficiency, despite the highly conserved structure of the active site. A. variabilis C503S/C565S PAL is shown to be both more thermally stable and more resistant to proteolytic cleavage than R. toruloides PAL. Additional increases in thermal stability and protease resistance upon ligand binding may be due to enhanced interactions among the residues of the active site, possibly locking the active-site structure in place and stabilizing the tetramer. Examination of the A. variabilis C503S/C565S PAL structure, combined with analysis of its physical properties, provides

  9. Synthesis of specifically labelled L-phenylalanines using phenylalanine ammonia lyase activity

    Energy Technology Data Exchange (ETDEWEB)

    Haedener, A.; Tamm, Ch.

    1987-11-01

    Specifically labelled L-phenylalanines have been prepared using a variety of classical synthetic methods in combination with phenylalanine ammonia lyase (PAL) enzyme activity of the yeast Rhodosporidium toruloides ATCC 10788 or Rhodotorula glutinis IFO 0559, respectively. Thus, L-(2-/sup 2/H)phenyl-(2-/sup 2/H)alanine was formed from (E) -(2,2'-/sup 2/H/sub 2/)cinnamic acid and ammonia in 46% yield, whereas L-phenyl-(2-/sup 13/C, /sup 15/N)alanine was obtained from (E)-(2-/sup 13/C)cinnamic acid in 45% overall yield. Generally, labelled cinnamic acids were recovered in pure form from the reaction mixture, with a loss of 6-8%. Likewise, unchanged /sup 15/NH/sub 3/ was reisolated as /sup 15/NH/sub 4/Cl after steam distillation with overall losses of less than 4%. Labelled cinnamic acids were prepared by Knoevenagel condensations between appropriately labelled benzaldehydes and malonic acids. (2-/sup 2/H)Benzaldehyde was obtained from 2-bromotoluene by decomposition of the corresponding Grignard reagent with /sup 2/H/sub 2/O and subsequent oxidation. Since simple molecules, most of them commercially available in labelled form or otherwise easily accessible, may serve as starting material, and due to its defined stereochemistry, the reaction catalysed by PAL opens a short and attractive route to specifically labelled L-phenylalanines.

  10. Formulation and PEGylation optimization of the therapeutic PEGylated phenylalanine ammonia lyase for the treatment of phenylketonuria.

    Science.gov (United States)

    Bell, Sean M; Wendt, Dan J; Zhang, Yanhong; Taylor, Timothy W; Long, Shinong; Tsuruda, Laurie; Zhao, Bin; Laipis, Phillip; Fitzpatrick, Paul A

    2017-01-01

    Phenylketonuria (PKU) is a genetic metabolic disease in which the decrease or loss of phenylalanine hydroxylase (PAH) activity results in elevated, neurotoxic levels of phenylalanine (Phe). Due to many obstacles, PAH enzyme replacement therapy is not currently an option. Treatment of PKU with an alternative enzyme, phenylalanine ammonia lyase (PAL), was first proposed in the 1970s. However, issues regarding immunogenicity, enzyme production and mode of delivery needed to be overcome. Through the evaluation of PAL enzymes from multiple species, three potential PAL enzymes from yeast and cyanobacteria were chosen for evaluation of their therapeutic potential. The addition of polyethylene glycol (PEG, MW = 20,000), at a particular ratio to modify the protein surface, attenuated immunogenicity in an animal model of PKU. All three PEGylated PAL candidates showed efficacy in a mouse model of PKU (BTBR Pahenu2) upon subcutaneous injection. However, only PEGylated Anabaena variabilis (Av) PAL-treated mice demonstrated sustained low Phe levels with weekly injection and was the only PAL evaluated that maintained full enzymatic activity upon PEGylation. A PEGylated recombinant double mutant version of AvPAL (Cys503Ser/Cys565Ser), rAvPAL-PEG, was selected for drug development based on its positive pharmacodynamic profile and favorable expression titers. PEGylation was shown to be critical for rAvPAL-PEG efficacy as under PEGylated rAvPAL had a lower pharmacodynamic effect. rAvPAL and rAvPAL-PEG had poor stability at 4°C. L-Phe and trans-cinnamate were identified as activity stabilizing excipients. rAvPAL-PEG is currently in Phase 3 clinical trials to assess efficacy in PKU patients.

  11. Formulation and PEGylation optimization of the therapeutic PEGylated phenylalanine ammonia lyase for the treatment of phenylketonuria.

    Directory of Open Access Journals (Sweden)

    Sean M Bell

    Full Text Available Phenylketonuria (PKU is a genetic metabolic disease in which the decrease or loss of phenylalanine hydroxylase (PAH activity results in elevated, neurotoxic levels of phenylalanine (Phe. Due to many obstacles, PAH enzyme replacement therapy is not currently an option. Treatment of PKU with an alternative enzyme, phenylalanine ammonia lyase (PAL, was first proposed in the 1970s. However, issues regarding immunogenicity, enzyme production and mode of delivery needed to be overcome. Through the evaluation of PAL enzymes from multiple species, three potential PAL enzymes from yeast and cyanobacteria were chosen for evaluation of their therapeutic potential. The addition of polyethylene glycol (PEG, MW = 20,000, at a particular ratio to modify the protein surface, attenuated immunogenicity in an animal model of PKU. All three PEGylated PAL candidates showed efficacy in a mouse model of PKU (BTBR Pahenu2 upon subcutaneous injection. However, only PEGylated Anabaena variabilis (Av PAL-treated mice demonstrated sustained low Phe levels with weekly injection and was the only PAL evaluated that maintained full enzymatic activity upon PEGylation. A PEGylated recombinant double mutant version of AvPAL (Cys503Ser/Cys565Ser, rAvPAL-PEG, was selected for drug development based on its positive pharmacodynamic profile and favorable expression titers. PEGylation was shown to be critical for rAvPAL-PEG efficacy as under PEGylated rAvPAL had a lower pharmacodynamic effect. rAvPAL and rAvPAL-PEG had poor stability at 4°C. L-Phe and trans-cinnamate were identified as activity stabilizing excipients. rAvPAL-PEG is currently in Phase 3 clinical trials to assess efficacy in PKU patients.

  12. Structure and characterization of a cDNA clone for phenylalanine ammonia-lyase from cut-injured roots of sweet potato

    International Nuclear Information System (INIS)

    Tanaka, Yoshiyuki; Matsuoka, Makoto; Yamanoto, Naoki; Ohashi, Yuko; Kano-Murakami, Yuriko; Ozeki, Yoshihiro

    1989-01-01

    A cDNA clone for phenylalanine ammonia-lyase (PAL) induced in wounded sweet potato (Ipomoea batatas Lam.) root was obtained by immunoscreening a cDNA library. The protein produced in Escherichia coli cells containing the plasmid pPAL02 was indistinguishable from sweet potato PAL as judged by Ouchterlony double diffusion assays. The M r of its subunit was 77,000. The cells converted [ 14 C]-L-phenylalanine into [ 14 C]-t-cinnamic acid and PAL activity was detected in the homogenate of the cells. The activity was dependent on the presence of the pPAL02 plasmid DNA. The nucleotide sequence of the cDNA contained a 2,121-base pair (bp) open-reading frame capable of coding for a polypeptide with 707 amino acids (M r 77,137), a 22-bp 5'-noncoding region and a 207-bp 3'-noncoding region. The results suggest that the insert DNA fully encoded the amino acid sequence for sweet potato PAL that is induced by wounding. Comparison of the deduced amino acid sequence with that of a PAL cDNA fragment from Phaseolus vulgaris revealed 78.9% homology. The sequence from amino acid residues 258 to 494 was highly conserved, showing 90.7% homology

  13. One-Pot Enzymatic Synthesis of D-Arylalanines Using Phenylalanine Ammonia Lyase and L-Amino Acid Deaminase.

    Science.gov (United States)

    Zhu, Longbao; Feng, Guoqiang; Ge, Fei; Song, Ping; Wang, Taotao; Liu, Yi; Tao, Yugui; Zhou, Zhemin

    2018-06-08

    The phenylalanine ammonia-lyase (AvPAL) from Anabaena variabilis catalyzes the amination of substituent trans-cinnamic acid (t-CA) to produce racemic D,L-enantiomer arylalanine mixture owing to its low stereoselectivity. To produce high optically pure D-arylalanine, a modified AvPAL with high D-selectivity is expected. Based on the analyses of catalytic mechanism and structure, the Asn347 residue in the active site was proposed to control stereoselectivity. Therefore, Asn347 was mutated to construct mutant AvPAL-N347A, the stereoselectivity of AvPAL-N347A for D-enantiomer arylalanine was 2.3-fold higher than that of wild-type AvPAL (WtPAL). Furthermore, the residual L-enantiomer product in reaction solution could be converted into the D-enantiomer product through stereoselective oxidation by PmLAAD and nonselective reduction by reducing agent NH 3 BH 3 . At optimal conditions, the conversion rate of t-CA and optical purity (enantiomeric excess (ee D )) of D-phenylalanine reached 82% and exceeded 99%, respectively. The two enzymes displayed activity toward a broad range of substrate and could be used to efficiently synthesize D-arylalanine with different groups on the phenyl ring. Among these D-arylalanines, the yield of m-nitro-D-phenylalanine was highest and reached 96%, and the ee D exceeded 99%. This one-pot synthesis using AvPAL and PmLAAD has prospects for industrial application.

  14. Genome-wide characterization of phenylalanine ammonia-lyase gene family in watermelon (Citrullus lanatus).

    Science.gov (United States)

    Dong, Chun-Juan; Shang, Qing-Mao

    2013-07-01

    Phenylalanine ammonia-lyase (PAL), the first enzyme in the phenylpropanoid pathway, plays a critical role in plant growth, development, and adaptation. PAL enzymes are encoded by a gene family in plants. Here, we report a genome-wide search for PAL genes in watermelon. A total of 12 PAL genes, designated ClPAL1-12, are identified . Nine are arranged in tandem in two duplication blocks located on chromosomes 4 and 7, and the other three ClPAL genes are distributed as single copies on chromosomes 2, 3, and 8. Both the cDNA and protein sequences of ClPALs share an overall high identity with each other. A phylogenetic analysis places 11 of the ClPALs into a separate cucurbit subclade, whereas ClPAL2, which belongs to neither monocots nor dicots, may serve as an ancestral PAL in plants. In the cucurbit subclade, seven ClPALs form homologous pairs with their counterparts from cucumber. Expression profiling reveals that 11 of the ClPAL genes are expressed and show preferential expression in the stems and male and female flowers. Six of the 12 ClPALs are moderately or strongly expressed in the fruits, particularly in the pulp, suggesting the potential roles of PAL in the development of fruit color and flavor. A promoter motif analysis of the ClPAL genes implies redundant but distinctive cis-regulatory structures for stress responsiveness. Finally, duplication events during the evolution and expansion of the ClPAL gene family are discussed, and the relationships between the ClPAL genes and their cucumber orthologs are estimated.

  15. Evaluation of pal activity, phenolic and flavonoid contents in three pistachio ( Pistacia vera L. ) cultivars grafted onto three different rootstocks

    OpenAIRE

    Nadernejad, N.; Ahmadimoghadam, A.; Hossyinifard, J.; Poorseyedi, S.

    2013-01-01

    Phenylalanine ammonia lyase (PAL) is a biochemical marker of the environmental stress and plays a pivotal role in phenolic synthesis. The lower ROS level and oxidative damage was observed in grafted plants and the rootstocks have a profound influence on the biochemical composition, especially phenolic compounds. Regarding the importance of the effect rootstocks have on scion in pistachio trees, this study was carried out to assess and compare three pistachio cultivars ("Ahmadaghaii", "Ohadi" ...

  16. Synthesis of d‐ and l‐Phenylalanine Derivatives by Phenylalanine Ammonia Lyases: A Multienzymatic Cascade Process†

    Science.gov (United States)

    Parmeggiani, Fabio; Lovelock, Sarah L.; Weise, Nicholas J.; Ahmed, Syed T.

    2015-01-01

    Abstract The synthesis of substituted d‐phenylalanines in high yield and excellent optical purity, starting from inexpensive cinnamic acids, has been achieved with a novel one‐pot approach by coupling phenylalanine ammonia lyase (PAL) amination with a chemoenzymatic deracemization (based on stereoselective oxidation and nonselective reduction). A simple high‐throughput solid‐phase screening method has also been developed to identify PALs with higher rates of formation of non‐natural d‐phenylalanines. The best variants were exploited in the chemoenzymatic cascade, thus increasing the yield and ee value of the d‐configured product. Furthermore, the system was extended to the preparation of those l‐phenylalanines which are obtained with a low ee value using PAL amination. PMID:27478261

  17. Synthesis of d- and l-Phenylalanine Derivatives by Phenylalanine Ammonia Lyases: A Multienzymatic Cascade Process**

    Science.gov (United States)

    Parmeggiani, Fabio; Lovelock, Sarah L; Weise, Nicholas J; Ahmed, Syed T; Turner, Nicholas J

    2015-01-01

    The synthesis of substituted d-phenylalanines in high yield and excellent optical purity, starting from inexpensive cinnamic acids, has been achieved with a novel one-pot approach by coupling phenylalanine ammonia lyase (PAL) amination with a chemoenzymatic deracemization (based on stereoselective oxidation and nonselective reduction). A simple high-throughput solid-phase screening method has also been developed to identify PALs with higher rates of formation of non-natural d-phenylalanines. The best variants were exploited in the chemoenzymatic cascade, thus increasing the yield and ee value of the d-configured product. Furthermore, the system was extended to the preparation of those l-phenylalanines which are obtained with a low ee value using PAL amination. PMID:25728350

  18. Circadian Rhythmicity in the Activities of Phenylalanine Ammonia-Lyase from Lemna perpusilla and Spirodela polyrhiza 1

    Science.gov (United States)

    Gordon, William R.; Koukkari, Willard L.

    1978-01-01

    The oscillations in phenylalanine ammonia-lyase activity from Spirodela polyrhiza and phenylalanine ammonia-lyase and tyrosine ammonia-lyase activities from Lemna perpusilla displayed a circadian rhythm under continuous light. Rhythmicity in enzymic activity could not be detected in continuous darkness since under this condition phenylalanine ammonia-lyase activity remains at a fairly constantly low level. Results from our studies of the oscillatory pattern of the respective activities of phenylalanine and tyrosine ammonia-lyase support their “inseparability.” PMID:16660569

  19. Ammonia lyases and aminomutases as biocatalysts for the synthesis of α-amino and β-amino acids.

    Science.gov (United States)

    Turner, Nicholas J

    2011-04-01

    Ammonia lyases catalyse the reversible addition of ammonia to cinnamic acid (1: R=H) and p-hydroxycinnamic (1: R=OH) to generate L-phenylalanine (2: R=H) and L-tyrosine (2: R=OH) respectively (Figure 1a). Both phenylalanine ammonia lyase (PAL) and tyrosine ammonia lyase (TAL) are widely distributed in plants, fungi and prokaryotes. Recently there has been interest in the use of these enzymes for the synthesis of a broader range of L-arylalanines. Aminomutases catalyse a related reaction, namely the interconversion of α-amino acids to β-amino acids (Figure 1b). In the case of L-phenylalanine, this reaction is catalysed by phenylalanine aminomutase (PAM) and proceeds stereospecifically via the intermediate cinnamic acid to generate β-Phe 3. Ammonia lyases and aminomutases are related in sequence and structure and share the same active site cofactor 4-methylideneimidazole-5-one (MIO). There is currently interest in the possibility of using these biocatalysts to prepare a wide range of enantiomerically pure l-configured α-amino and β-amino acids. Recent reviews have focused on the mechanism of these MIO containing enzymes. The aim of this review is to review recent progress in the application of ammonia lyase and aminomutase enzymes to prepare enantiomerically pure α-amino and β-amino acids. Copyright © 2010 Elsevier Ltd. All rights reserved.

  20. Phenylalanine ammonia lyase catalyzed synthesis of amino acids by an MIO-cofactor independent pathway.

    Science.gov (United States)

    Lovelock, Sarah L; Lloyd, Richard C; Turner, Nicholas J

    2014-04-25

    Phenylalanine ammonia lyases (PALs) belong to a family of 4-methylideneimidazole-5-one (MIO) cofactor dependent enzymes which are responsible for the conversion of L-phenylalanine into trans-cinnamic acid in eukaryotic and prokaryotic organisms. Under conditions of high ammonia concentration, this deamination reaction is reversible and hence there is considerable interest in the development of PALs as biocatalysts for the enantioselective synthesis of non-natural amino acids. Herein the discovery of a previously unobserved competing MIO-independent reaction pathway, which proceeds in a non-stereoselective manner and results in the generation of both L- and D-phenylalanine derivatives, is described. The mechanism of the MIO-independent pathway is explored through isotopic-labeling studies and mutagenesis of key active-site residues. The results obtained are consistent with amino acid deamination occurring by a stepwise E1 cB elimination mechanism. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  1. Biochemical Evaluation of Phenylalanine Ammonia Lyase from Endemic Plant Cyathobasis fruticulosa (Bunge Aellen. for the Dietary Treatment of Phenylketonuria

    Directory of Open Access Journals (Sweden)

    Seda Şirin

    2016-01-01

    Full Text Available Enzyme substitution therapy with the phenylalanine ammonia lyase (PAL is a new approach to the treatment of patients with phenylketonuria (PKU. This enzyme is responsible for the conversion of phenylalanine to trans-cinnamic acid. We assessed the PAL enzyme of the endemic plant Cyathobasis fruticulosa (Bunge Aellen. for its possible role in the dietary treatment of PKU. The enzyme was found to have a high activity of (64.9±0.1 U/mg, with the optimum pH, temperature and buffer (Tris–HCl and L-phenylalanine concentration levels of pH=8.8, 37 °C and 100 mM, respectively. Optimum enzyme activity was achieved at pH=4.0 and 7.5, corresponding to pH levels of gastric and intestinal juice, and NaCl concentration of 200 mM. The purifi cation of the enzyme by 1.87-fold yielded an activity of 98.6 U/mg. PAL activities determined by HPLC analyses before and after purification were similar. Two protein bands, one at 70 and the other at 23 kDa, were determined by Western blot analysis of the enzyme. This enzyme is a potential candidate for serial production of dietary food and biotechnological products.

  2. Cloning and Expression Analysis of Phenylalanine Ammonia-Lyase Gene in the Mycelium and Fruit Body of the Edible Mushroom Flammulina velutipes

    Science.gov (United States)

    Yun, Yeo Hong; Koo, Ja Sun

    2015-01-01

    Phenylalanine ammonia-lyase (PAL) gene is known to be expressed in plants, and is involved in the differentiation, growth and synthesis of secondary metabolites. However, its expression in fungi remains to be explored. To understand its expression in mushroom fungi, the PAL gene of the edible mushroom Flammulina velutipes (Fvpal) was cloned and characterized. The cloned Fvpal consists of 2,175 bp, coding for a polypeptide containing 724 amino acids and having 11 introns. The translated amino acid sequence of Fvpal shares a high identity (66%) with that of ectomycorrhizal fungus Tricholoma matsutake. Distinctively, the Fvpal expression in the mycelium was higher in minimal medium supplemented with L-tyrosine than with other aromatic amino acids. During cultivation of the mushroom on sawdust medium, Fvpal expression in the fruit body correspondingly increased as the mushroom grew. In the fruiting body, Fvpal was expressed more in the stipe than in the pileus. These results suggest that F. velutipes PAL activity differs in the different organs of the mushroom. Overall, this is first report to show that the PAL gene expression is associated with mushroom growth in fungi. PMID:26539050

  3. Single-dose, subcutaneous recombinant phenylalanine ammonia lyase conjugated with polyethylene glycol in adult patients with phenylketonuria: an open-label, multicentre, phase 1 dose-escalation trial.

    Science.gov (United States)

    Longo, Nicola; Harding, Cary O; Burton, Barbara K; Grange, Dorothy K; Vockley, Jerry; Wasserstein, Melissa; Rice, Gregory M; Dorenbaum, Alejandro; Neuenburg, Jutta K; Musson, Donald G; Gu, Zhonghua; Sile, Saba

    2014-07-05

    Phenylketonuria is an inherited disease caused by impaired activity of phenylalanine hydroxylase, the enzyme that converts phenylalanine to tyrosine, leading to accumulation of phenylalanine and subsequent neurocognitive dysfunction. Phenylalanine ammonia lyase is a prokaryotic enzyme that converts phenylalanine to ammonia and trans-cinnamic acid. We aimed to assess the safety, tolerability, pharmacokinetic characteristics, and efficacy of recombinant Anabaena variabilis phenylalanine ammonia lyase (produced in Escherichia coli) conjugated with polyethylene glycol (rAvPAL-PEG) in reducing phenylalanine concentrations in adult patients with phenylketonuria. In this open-label, phase 1, multicentre trial, single subcutaneous injections of rAvPAL-PEG were given in escalating doses (0·001, 0·003, 0·010, 0·030, and 0·100 mg/kg) to adults with phenylketonuria. Participants aged 18 years or older with blood phenylalanine concentrations of 600 μmol/L or higher were recruited from among patients attending metabolic disease clinics in the USA. The primary endpoints were safety and tolerability of rAvPAL-PEG. Secondary endpoints were the pharmacokinetic characteristics of the drug and its effect on concentrations of phenylalanine. Participants and investigators were not masked to assigned dose group. This study is registered with ClinicalTrials.gov, number NCT00925054. 25 participants were recruited from seven centres between May 6, 2008, and April 15, 2009, with five participants assigned to each escalating dose group. All participants were included in the safety population. The most frequently reported adverse events were injection-site reactions and dizziness, which were self-limited and without sequelae. Two participants had serious adverse reactions to intramuscular medroxyprogesterone acetate, a drug that contains polyethylene glycol as an excipient. Three of five participants given the highest dose of rAvPAL-PEG (0·100 mg/kg) developed a generalised skin rash

  4. Phase 1 Trial of Subcutaneous rAvPAL-PEG in Subjects with Phenylketonuria

    Science.gov (United States)

    Longo, Nicola; Harding, Cary O.; Burton, Barbara K.; Grange, Dorothy K.; Vockley, Jerry; Wasserstein, Melissa; Rice, Gregory M.; Musson, Donald G.; Gu, Zhonghua; Sile, Saba

    2014-01-01

    Objective Phenylketonuria is an inherited disease caused by impaired activity of phenylalanine hydroxylase, the enzyme that converts phenylalanine to tyrosine, leading to accumulation of phenylalanine and subsequent neurocognitive dysfunction. A phenylalanine-restricted diet initiated early in life can ameliorate the toxic effects of phenylalanine. However, the diet is onerous and compliance is extremely difficult. Phenylalanine ammonia lyase (PAL) is a prokaryotic enzyme that converts phenylalanine to ammonia and trans-cinnamic acid. This Phase 1, multicenter clinical trial evaluated the safety, tolerability, pharmacokinetics and efficacy of rAvPAL-PEG (recombinant Anabaena variabilis PAL produced in E. coli conjugated with polyethylene glycol [PEG] to reduce immunogenicity) in reducing phenylalanine levels in subjects with phenylketonuria. Methods Single subcutaneous injections of rAvPAL-PEG in escalating doses (0·001, 0·003, 0·01, 0·03, and 0·1 mg/kg) were administered to 25 adults with phenylketonuria recruited from those attending metabolic clinics in North America whose blood phenylalanine concentrations were ≥600 μmol/L. Results The most frequently reported adverse events were injection-site reactions and dizziness. Reactions were self-limited without sequelae. During the trial, two subjects had adverse reactions to intramuscular (IM) medroxyprogesterone acetate, a drug containing polyethylene glycol as an excipient. Three subjects developed a generalized skin rash at the highest rAvPAL-PEG dose (0·1 mg/kg). Drug levels peaked ∼5 days after the injection. Treatment was effective in reducing blood phenylalanine in all five subjects receiving the highest dose (0·1 mg/kg, mean percent change of -58 from baseline), with a nadir ∼6 days after injection and inverse correlation between drug and phenylalanine concentrations in plasma. Phenylalanine concentrations returned to near-baseline levels ∼20 days after the single injection. Conclusions

  5. DNA Methylation Influences Chlorogenic Acid Biosynthesis in Lonicera japonica by Mediating LjbZIP8 to Regulate Phenylalanine Ammonia-Lyase 2 Expression

    Directory of Open Access Journals (Sweden)

    Liangping Zha

    2017-07-01

    Full Text Available The content of active compounds differ in buds and flowers of Lonicera japonica (FLJ and L. japonica var. chinensis (rFLJ. Chlorogenic acid (CGAs were major active compounds of L. japonica and regarded as measurements for quality evaluation. However, little is known concerning the formation of active compounds at the molecular level. We quantified the major CGAs in FLJ and rFLJ, and found the concentrations of CGAs were higher in the buds of rFLJ than those of FLJ. Further analysis of CpG methylation of CGAs biosynthesis genes showed differences between FLJ and rFLJ in the 5′-UTR of phenylalanine ammonia-lyase 2 (PAL2. We identified 11 LjbZIP proteins and 24 rLjbZIP proteins with conserved basic leucine zipper domains, subcellular localization, and electrophoretic mobility shift assay showed that the transcription factor LjbZIP8 is a nuclear-localized protein that specifically binds to the G-box element of the LjPAL2 5′-UTR. Additionally, a transactivation assay and LjbZIP8 overexpression in transgenic tobacco indicated that LjbZIP8 could function as a repressor of transcription. Finally, treatment with 5-azacytidine decreased the transcription level of LjPAL2 and CGAs content in FLJ leaves. These results raise the possibility that DNA methylation might influence the recruitment of LjbZIP8, regulating PAL2 expression level and CGAs content in L. japonica.

  6. Conservation, Divergence, and Genome-Wide Distribution of PAL and POX A Gene Families in Plants.

    Science.gov (United States)

    Rawal, H C; Singh, N K; Sharma, T R

    2013-01-01

    Genome-wide identification and phylogenetic and syntenic comparison were performed for the genes responsible for phenylalanine ammonia lyase (PAL) and peroxidase A (POX A) enzymes in nine plant species representing very diverse groups like legumes (Glycine max and Medicago truncatula), fruits (Vitis vinifera), cereals (Sorghum bicolor, Zea mays, and Oryza sativa), trees (Populus trichocarpa), and model dicot (Arabidopsis thaliana) and monocot (Brachypodium distachyon) species. A total of 87 and 1045 genes in PAL and POX A gene families, respectively, have been identified in these species. The phylogenetic and syntenic comparison along with motif distributions shows a high degree of conservation of PAL genes, suggesting that these genes may predate monocot/eudicot divergence. The POX A family genes, present in clusters at the subtelomeric regions of chromosomes, might be evolving and expanding with higher rate than the PAL gene family. Our analysis showed that during the expansion of POX A gene family, many groups and subgroups have evolved, resulting in a high level of functional divergence among monocots and dicots. These results will act as a first step toward the understanding of monocot/eudicot evolution and functional characterization of these gene families in the future.

  7. Conservation, Divergence, and Genome-Wide Distribution of PAL and POX A Gene Families in Plants

    Directory of Open Access Journals (Sweden)

    H. C. Rawal

    2013-01-01

    Full Text Available Genome-wide identification and phylogenetic and syntenic comparison were performed for the genes responsible for phenylalanine ammonia lyase (PAL and peroxidase A (POX A enzymes in nine plant species representing very diverse groups like legumes (Glycine max and Medicago truncatula, fruits (Vitis vinifera, cereals (Sorghum bicolor, Zea mays, and Oryza sativa, trees (Populus trichocarpa, and model dicot (Arabidopsis thaliana and monocot (Brachypodium distachyon species. A total of 87 and 1045 genes in PAL and POX A gene families, respectively, have been identified in these species. The phylogenetic and syntenic comparison along with motif distributions shows a high degree of conservation of PAL genes, suggesting that these genes may predate monocot/eudicot divergence. The POX A family genes, present in clusters at the subtelomeric regions of chromosomes, might be evolving and expanding with higher rate than the PAL gene family. Our analysis showed that during the expansion of POX A gene family, many groups and subgroups have evolved, resulting in a high level of functional divergence among monocots and dicots. These results will act as a first step toward the understanding of monocot/eudicot evolution and functional characterization of these gene families in the future.

  8. The Ve-mediated resistance response of the tomato to Verticillium dahliae involves H2O2, peroxidase and lignins and drives PAL gene expression

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    Merino Fuencisla

    2010-10-01

    Full Text Available Abstract Background Verticillium dahliae is a fungal pathogen that infects a wide range of hosts. The only known genes for resistance to Verticillium in the Solanaceae are found in the tomato (Solanum lycopersicum Ve locus, formed by two linked genes, Ve1 and Ve2. To characterize the resistance response mediated by the tomato Ve gene, we inoculated two nearly isogenic tomato lines, LA3030 (ve/ve and LA3038 (Ve/Ve, with V. dahliae. Results We found induction of H2O2 production in roots of inoculated plants, followed by an increase in peroxidase activity only in roots of inoculated resistant plants. Phenylalanine-ammonia lyase (PAL activity was also increased in resistant roots 2 hours after inoculation, while induction of PAL activity in susceptible roots was not seen until 48 hours after inoculation. Phenylpropanoid metabolism was also affected, with increases in ferulic acid, p-coumaric acid, vanillin and p-hydroxybenzaldehyde contents in resistant roots after inoculation. Six tomato PAL cDNA sequences (PAL1 - PAL6 were found in the SolGenes tomato EST database. RT-PCR analysis showed that these genes were expressed in all organs of the plant, albeit at different levels. Real-time RT-PCR indicated distinct patterns of expression of the different PAL genes in V. dahliae-inoculated roots. Phylogenetic analysis of 48 partial PAL cDNAs corresponding to 19 plant species grouped angiosperm PAL sequences into four clusters, suggesting functional differences among the six tomato genes, with PAL2 and PAL6 presumably involved in lignification, and the remaining PAL genes implicated in other biological processes. An increase in the synthesis of lignins was found 16 and 28 days after inoculation in both lines; this increase was greater and faster to develop in the resistant line. In both resistant and susceptible inoculated plants, an increase in the ratio of guaiacyl/syringyl units was detected 16 days after inoculation, resulting from the lowered amount

  9. Examining the validity of the ActivPAL monitor in measuring posture and ambulatory movement in children

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    Aminian Saeideh

    2012-10-01

    Full Text Available Abstract Background Decreasing sedentary activities that involve prolonged sitting may be an important strategy to reduce obesity and other physical and psychosocial health problems in children. The first step to understanding the effect of sedentary activities on children’s health is to objectively assess these activities with a valid measurement tool. Purpose To examine the validity of the ActivPAL monitor in measuring sitting/lying, standing, and walking time, transition counts and step counts in children in a laboratory setting. Methods Twenty five healthy elementary school children (age 9.9 ± 0.3 years; BMI 18.2 ± 1.9; mean ± SD were randomly recruited across the Auckland region, New Zealand. Children were fitted with ActivPAL monitors and observed during simulated free-living activities involving sitting/lying, standing and walking, followed by treadmill and over-ground activities at various speeds (slow, normal, fast against video observation (criterion measure. The ActivPAL sit-to-stand and stand-to-sit transition counts and steps were also compared with video data. The accuracy of step counts measured by the ActivPAL was also compared against the New Lifestyles NL-2000 and the Yamax Digi-Walker SW-200 pedometers. Results We observed a perfect correlation between the ActivPAL monitor in time spent sitting/lying, standing, and walking in simulated free-living activities with direct observation. Correlations between the ActivPAL and video observation in total numbers of sit-to-stand and stand-to-sit transitions were high (r = 0.99 ± 0.01. Unlike pedometers, the ActivPAL did not misclassify fidgeting as steps taken. Strong correlations (r = 0.88-1.00 between ActivPAL step counts and video observation in both treadmill and over-ground slow and normal walking were also observed. During treadmill and over-ground fast walking and running, the correlations were low (r = 0.21-0.46. Conclusion The ActivPAL monitor is

  10. Efficient preparation of enantiopure D-phenylalanine through asymmetric resolution using immobilized phenylalanine ammonia-lyase from Rhodotorula glutinis JN-1 in a recirculating packed-bed reactor.

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    Longbao Zhu

    Full Text Available An efficient enzymatic process was developed to produce optically pure D-phenylalanine through asymmetric resolution of the racemic DL-phenylalanine using immobilized phenylalanine ammonia-lyase (RgPAL from Rhodotorula glutinis JN-1. RgPAL was immobilized on a modified mesoporous silica support (MCM-41-NH-GA. The resulting MCM-41-NH-GA-RgPAL showed high activity and stability. The resolution efficiency using MCM-41-NH-GA-RgPAL in a recirculating packed-bed reactor (RPBR was higher than that in a stirred-tank reactor. Under optimal operational conditions, the volumetric conversion rate of L-phenylalanine and the productivity of D-phenylalanine reached 96.7 mM h⁻¹ and 0.32 g L⁻¹ h⁻¹, respectively. The optical purity (eeD of D-phenylalanine exceeded 99%. The RPBR ran continuously for 16 batches, the conversion ratio did not decrease. The reactor was scaled up 25-fold, and the productivity of D-phenylalanine (eeD>99% in the scaled-up reactor reached 7.2 g L⁻¹ h⁻¹. These results suggest that the resolution process is an alternative method to produce highly pure D-phenylalanine.

  11. Efficient preparation of enantiopure D-phenylalanine through asymmetric resolution using immobilized phenylalanine ammonia-lyase from Rhodotorula glutinis JN-1 in a recirculating packed-bed reactor.

    Science.gov (United States)

    Zhu, Longbao; Zhou, Li; Huang, Nan; Cui, Wenjing; Liu, Zhongmei; Xiao, Ke; Zhou, Zhemin

    2014-01-01

    An efficient enzymatic process was developed to produce optically pure D-phenylalanine through asymmetric resolution of the racemic DL-phenylalanine using immobilized phenylalanine ammonia-lyase (RgPAL) from Rhodotorula glutinis JN-1. RgPAL was immobilized on a modified mesoporous silica support (MCM-41-NH-GA). The resulting MCM-41-NH-GA-RgPAL showed high activity and stability. The resolution efficiency using MCM-41-NH-GA-RgPAL in a recirculating packed-bed reactor (RPBR) was higher than that in a stirred-tank reactor. Under optimal operational conditions, the volumetric conversion rate of L-phenylalanine and the productivity of D-phenylalanine reached 96.7 mM h⁻¹ and 0.32 g L⁻¹ h⁻¹, respectively. The optical purity (eeD) of D-phenylalanine exceeded 99%. The RPBR ran continuously for 16 batches, the conversion ratio did not decrease. The reactor was scaled up 25-fold, and the productivity of D-phenylalanine (eeD>99%) in the scaled-up reactor reached 7.2 g L⁻¹ h⁻¹. These results suggest that the resolution process is an alternative method to produce highly pure D-phenylalanine.

  12. AlgM4: A New Salt-Activated Alginate Lyase of the PL7 Family with Endolytic Activity

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    Guiyuan Huang

    2018-04-01

    Full Text Available Alginate lyases are a group of enzymes that catalyze the depolymerization of alginates into oligosaccharides or monosaccharides. These enzymes have been widely used for a variety of purposes, such as producing bioactive oligosaccharides, controlling the rheological properties of polysaccharides, and performing structural analyses of polysaccharides. The algM4 gene of the marine bacterium Vibrio weizhoudaoensis M0101 encodes an alginate lyase that belongs to the polysaccharide lyase family 7 (PL7. In this study, the kinetic constants Vmax (maximum reaction rate and Km (Michaelis constant of AlgM4 activity were determined as 2.75 nmol/s and 2.72 mg/mL, respectively. The optimum temperature for AlgM4 activity was 30 °C, and at 70 °C, AlgM4 activity dropped to 11% of the maximum observed activity. The optimum pH for AlgM4 activity was 8.5, and AlgM4 was completely inactive at pH 11. The addition of 1 mol/L NaCl resulted in a more than sevenfold increase in the relative activity of AlgM4. The secondary structure of AlgM4 was altered in the presence of NaCl, which caused the α-helical content to decrease from 12.4 to 10.8% and the β-sheet content to decrease by 1.7%. In addition, NaCl enhanced the thermal stability of AlgM4 and increased the midpoint of thermal denaturation (Tm by 4.9 °C. AlgM4 exhibited an ability to degrade sodium alginate, poly-mannuronic acid (polyM, and poly-guluronic acid (polyG, resulting in the production of oligosaccharides with a degree of polymerization (DP of 2–9. AlgM4 possessed broader substrate, indicating that it is a bifunctional alginate lyase. Thus, AlgM4 is a novel salt-activated and bifunctional alginate lyase of the PL7 family with endolytic activity.

  13. Genomic Characterization of Phenylalanine Ammonia Lyase Gene in Buckwheat.

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    Karthikeyan Thiyagarajan

    Full Text Available Phenylalanine Ammonia Lyase (PAL gene which plays a key role in bio-synthesis of medicinally important compounds, Rutin/quercetin was sequence characterized for its efficient genomics application. These compounds possessing anti-diabetic and anti-cancer properties and are predominantly produced by Fagopyrum spp. In the present study, PAL gene was sequenced from three Fagopyrum spp. (F. tataricum, F. esculentum and F. dibotrys and showed the presence of three SNPs and four insertion/deletions at intra and inter specific level. Among them, the potential SNP (position 949th bp G>C with Parsimony Informative Site was selected and successfully utilised to individuate the zygosity/allelic variation of 16 F. tataricum varieties. Insertion mutations were identified in coding region, which resulted the change of a stretch of 39 amino acids on the putative protein. Our Study revealed that autogamous species (F. tataricum has lower frequency of observed SNPs as compared to allogamous species (F. dibotrys and F. esculentum. The identified SNPs in F. tataricum didn't result to amino acid change, while in other two species it caused both conservative and non-conservative variations. Consistent pattern of SNPs across the species revealed their phylogenetic importance. We found two groups of F. tataricum and one of them was closely related with F. dibotrys. Sequence characterization information of PAL gene reported in present investigation can be utilized in genetic improvement of buckwheat in reference to its medicinal value.

  14. Processes for the production of hydroxycinnamic acids using polypeptides having tyrosine ammonia lyase activity

    DEFF Research Database (Denmark)

    2016-01-01

    The present invention generally relates to the field of biotechnology as it applies to the production of hydroxycinnamic acids using polypeptides having tyrosine ammonia lyase activity. More particularly, the present invention pertains to polypeptides having tyrosine ammonia lyase activity and high...... substrate specificity towards tyrosine, which makes them particularly suitable in the production of p-coumaric acid and other hydroxycinnamic acids. The present invention thus provides processes for the production of p-coumaric acid and other hydroxycinnamic acids employing these polypeptides as well...

  15. Phenylalanine Ammonia Lyase Activity in Stem of Pepper (Capsicum annuum L.) Infected by Phytophthora capsici L.

    OpenAIRE

    KOC, Esra; USTUN, Ayşen Sulun

    2012-01-01

    In this study, PAL activity in stems of pepper (Capsicum annuum L.) plants infected by the root rot pathogen Phytophthora capsici-22 in a resistant (PM-702) and two susceptible (Kahramanmaraş-Hot (KM-Hot) and Demre-8) cultivar were studied. The response of the PAL activity in the resistant cultivar was faster and higher than in the susceptible cultivars (p < 0.01). The increase in production of  PAL upon Phytophthora capsici-22 were higher in the infected plants compared to the non-in...

  16. Activities of methionine-γ-lyase in the acidophilic archaeon “Ferroplasma acidarmanus” strain fer1

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    Khan MA

    2013-04-01

    Full Text Available M A Khan,1 Madeline M López-Muñoz,2 Charles W Kaspar,3 Kai F Hung1 1Department of Biological Sciences, Eastern Illinois University, Charleston, IL, USA; 2Department of Biology, Universidad de Puerto Rico, Mayaguez, Puerto Rico; 3Bacteriology Department, University of Wisconsin, Madison, WI, USA Abstract: Biogeochemical processes on exposed pyrite ores result in extremely high levels of sulfuric acid at these locations. Acidophiles that thrive in these conditions must overcome significant challenges, including an environment with proton concentrations at pH 3 or below. The role of sulfur metabolism in the archaeon “Ferroplasma acidarmanus” strain fer1's ability to thrive in this environment was investigated due to its growth-dependent production of methanethiol, a volatile organic sulfur compound. Two putative sequences for methionine-γ-lyase (EC 4.4.1.11, an enzyme known to carry out α, γ-elimination on L-methionine to produce methanethiol, were identified in fer1. Bioinformatic analyses identified a conserved pyridoxal-5'-phosphate (PLP binding domain and a partially conserved catalytic domain in both putative sequences. Detection of PLP-dependent and L-methionine-dependent production of α-keto compounds and thiol groups in fer1 confirmed the presence of methionine-γ-lyase activity. Further, fer1 lysate was capable of processing related substrates, including D-methionine, L-cysteine, L-cystathionine, and L/D-homocysteine. When the two putative fer1 methionine-γ-lyase gene-coded proteins were expressed in Escherichia coli cells, one sequence demonstrated an ability to carry out α, γ-elimination activity, while the other exhibited γ-replacement activity. These fer1 methionine-γ-lyases also exhibited optimum pH, substrate specificity, and catalytic preferences that are different from methionine-γ-lyases from other organisms. These differences are discussed in the context of molecular phylogeny constructed using a maximum

  17. Hydrogen Peroxide Treatment and the Phenylpropanoid Pathway Precursors Feeding Improve Phenolics and Antioxidant Capacity of Quinoa Sprouts via an Induction of L-Tyrosine and L-Phenylalanine Ammonia-Lyases Activities

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    Michał Świeca

    2016-01-01

    Full Text Available Hydrogen peroxide treatment and the phenylpropanoid pathway precursors feeding affected the antioxidant capacity of quinoa sprouts. Compared to the control, total phenolics content was significantly increased by treatment of control sprouts with 50 mM and 200 mM H2O2—an elevation of about 24% and 28%, respectively. The highest increase of flavonoids content was found for the sprouts treated with 200 mM H2O2 obtained from seeds fed with shikimic acid. All the studied modifications increased the antioxidant potential of sprouts (at least by 50% compared to control. The highest reducing power was found for the sprouts treated with 200 mM H2O2 obtained by phenylalanine feeding (5.03 mg TE/g DW and those obtained from the seeds fed with tyrosine (5.26 mg TE/g DW. The activities of L-tyrosine (TAL and L-phenylalanine (PAL ammonia-lyases were strongly affected by germination time as well as the applied modification of sprouting. On the 3rd day the highest PAL activity was determined for both untreated and induced with 50 mM H2O2 sprouts obtained by phenylalanine feeding. H2O2 induced TAL activity; the highest TAL activity was determined for 3-day-old sprouts induced with 200 mM H2O2 obtained from seeds fed with phenylalanine.

  18. Soumen Pal

    Indian Academy of Sciences (India)

    Home; Journals; Bulletin of Materials Science. Soumen Pal. Articles written in Bulletin of Materials Science. Volume 32 Issue 2 April 2009 pp 169-176 Composites. Sintering behaviour of spinel–alumina composites · Soumen Pal A K Bandyopadhyay P G Pal S Mukherjee B N Samaddar · More Details Abstract Fulltext PDF.

  19. Understanding Which Residues of the Active Site and Loop Structure of a Tyrosine Aminomutase Define Its Mutase and Lyase Activities.

    Science.gov (United States)

    Attanayake, Gayanthi; Walter, Tyler; Walker, Kevin D

    2018-05-30

    Site-directed mutations and substrate analogues were used to gain insights into the branch-point reaction of the 3,5-dihydro-5-methylidene-4 H-imidazol-4-one (MIO)-tyrosine aminomutase from Oryza sativa ( OsTAM). Exchanging the active residues of OsTAM (Y125C/N446K) for those in a phenylalanine aminomutase TcPAM altered its substrate specificity from tyrosine to phenylalanine. The aminomutase mechanism of OsTAM surprisingly changed almost exclusively to that of an ammonia lyase making cinnamic acid (>95%) over β-phenylalanine [Walter, T., et al. (2016) Biochemistry 55, 3497-3503]. We hypothesized that the missing electronics or sterics on the aryl ring of the phenylalanine substrate, compared with the sizable electron-donating hydroxyl of the natural tyrosine substrate, influenced the unexpected lyase reactivity of the OsTAM mutant. The double mutant was incubated with 16 α-phenylalanine substituent analogues of varying electronic strengths and sterics. The mutant converted each analogue principally to its acrylate with ∼50% conversion of the p-Br substrate, making only a small amount of the β-amino acid. The inner loop structure over the entrance to the active site was also mutated to assess how the lyase and mutase activities are affected. An OsTAM loop mutant, matching the loop residues of TcPAM, still chiefly made >95% of the acrylate from each substrate. A combined active site:loop mutant was most reactive but remained a lyase, making 10-fold more acrylates than other mutants did. While mutations within the active site changed the substrate specificity of OsTAM, continued exploration is needed to fully understand the interplay among the inner loop, the substrate, and the active site in defining the mutase and lyase activities.

  20. Structure-based functional annotation of putative conserved proteins having lyase activity from Haemophilus influenzae.

    Science.gov (United States)

    Shahbaaz, Mohd; Ahmad, Faizan; Imtaiyaz Hassan, Md

    2015-06-01

    Haemophilus influenzae is a small pleomorphic Gram-negative bacteria which causes several chronic diseases, including bacteremia, meningitis, cellulitis, epiglottitis, septic arthritis, pneumonia, and empyema. Here we extensively analyzed the sequenced genome of H. influenzae strain Rd KW20 using protein family databases, protein structure prediction, pathways and genome context methods to assign a precise function to proteins whose functions are unknown. These proteins are termed as hypothetical proteins (HPs), for which no experimental information is available. Function prediction of these proteins would surely be supportive to precisely understand the biochemical pathways and mechanism of pathogenesis of Haemophilus influenzae. During the extensive analysis of H. influenzae genome, we found the presence of eight HPs showing lyase activity. Subsequently, we modeled and analyzed three-dimensional structure of all these HPs to determine their functions more precisely. We found these HPs possess cystathionine-β-synthase, cyclase, carboxymuconolactone decarboxylase, pseudouridine synthase A and C, D-tagatose-1,6-bisphosphate aldolase and aminodeoxychorismate lyase-like features, indicating their corresponding functions in the H. influenzae. Lyases are actively involved in the regulation of biosynthesis of various hormones, metabolic pathways, signal transduction, and DNA repair. Lyases are also considered as a key player for various biological processes. These enzymes are critically essential for the survival and pathogenesis of H. influenzae and, therefore, these enzymes may be considered as a potential target for structure-based rational drug design. Our structure-function relationship analysis will be useful to search and design potential lead molecules based on the structure of these lyases, for drug design and discovery.

  1. Studies on pectin lyase

    NARCIS (Netherlands)

    Houdenhoven, van F.E.A.

    1975-01-01

    The pectin lyase activity in the commercial enzyme preparation Ultrazym originates from more then one type of enzyme; two of them, accounting for 95 % of the total activity, have been completely purified. As purity criteria specific activity, polyacrylamide disc gel electrophoresis and SDS

  2. META-ANALYSIS OF PHYSICAL ACTIVITY LEVEL ( PAL ) DATA FOR U.S. YOUTH

    Science.gov (United States)

    This paper describes analytical research undertaken by EMRB staff to obtain age- and gender-specific distributions of Physical Activity Level (PAL) values for children measured in studies published between 1990-2004 in the clinical nutrition and exercise physiology literatures. ...

  3. Validation of the Actigraph GT3X and ActivPAL Accelerometers for the Assessment of Sedentary Behavior

    Science.gov (United States)

    Kim, Youngdeok; Barry, Vaughn W.; Kang, Minsoo

    2015-01-01

    This study examined (a) the validity of two accelerometers (ActiGraph GT3X [ActiGraph LLC, Pensacola, FL, USA] and activPAL [PAL Technologies Ltd., Glasgow, Scotland]) for the assessment of sedentary behavior; and (b) the variations in assessment accuracy by setting minimum sedentary bout durations against a proxy for direct observation using an…

  4. Molecular characterization of a Penicillium chrysogenum exo-rhamnogalacturonan lyase that is structurally distinct from other polysaccharide lyase family proteins.

    Science.gov (United States)

    Iwai, Marin; Kawakami, Takuya; Ikemoto, Takeshi; Fujiwara, Daisuke; Takenaka, Shigeo; Nakazawa, Masami; Ueda, Mitsuhiro; Sakamoto, Tatsuji

    2015-10-01

    We previously described an endo-acting rhamnogalacturonan (RG) lyase, termed PcRGL4A, of Penicillium chrysogenum 31B. Here, we describe a second RG lyase, called PcRGLX. We determined the cDNA sequence of the Pcrglx gene, which encodes PcRGLX. Based on analyses using a BLAST search and a conserved domain search, PcRGLX was found to be structurally distinct from known RG lyases and might belong to a new polysaccharide lyase family together with uncharacterized fungal proteins of Nectria haematococca, Aspergillus oryzae, and Fusarium oxysporum. The Pcrglx cDNA gene product (rPcRGLX) expressed in Escherichia coli demonstrated specific activity against RG but not against homogalacturonan. Divalent cations were not essential for the enzymatic activity of rPcRGLX. rPcRGLX mainly released unsaturated galacturonosyl rhamnose (ΔGR) from RG backbones used as the substrate from the initial stage of the reaction, indicating that the enzyme can be classified as an exo-acting RG lyase (EC 4.2.2.24). This is the first report of an RG lyase with this mode of action in Eukaryota. rPcRGLX acted synergistically with PcRGL4A to degrade soybean RG and released ΔGR. This ΔGR was partially decorated with galactose (Gal) residues, indicating that rPcRGLX preferred oligomeric RGs to polymeric RGs, that the enzyme did not require Gal decoration of RG backbones for degradation, and that the enzyme bypassed the Gal side chains of RG backbones. These characteristics of rPcRGLX might be useful in the determination of complex structures of pectins.

  5. Sitaram Pal

    Indian Academy of Sciences (India)

    Home; Journals; Pramana – Journal of Physics. Sitaram Pal. Articles written in Pramana – Journal of Physics. Volume 68 Issue 5 May 2007 pp 789-801 Research Articles. Evidence of self-affne multiplicity scaling of charged-particle multiplicity distribution in hadron–nucleus interaction · Dipak Ghosh Argha Deb Sitaram Pal ...

  6. Monovalent Cation Activation of the Radical SAM Enzyme Pyruvate Formate-Lyase Activating Enzyme.

    Science.gov (United States)

    Shisler, Krista A; Hutcheson, Rachel U; Horitani, Masaki; Duschene, Kaitlin S; Crain, Adam V; Byer, Amanda S; Shepard, Eric M; Rasmussen, Ashley; Yang, Jian; Broderick, William E; Vey, Jessica L; Drennan, Catherine L; Hoffman, Brian M; Broderick, Joan B

    2017-08-30

    Pyruvate formate-lyase activating enzyme (PFL-AE) is a radical S-adenosyl-l-methionine (SAM) enzyme that installs a catalytically essential glycyl radical on pyruvate formate-lyase. We show that PFL-AE binds a catalytically essential monovalent cation at its active site, yet another parallel with B 12 enzymes, and we characterize this cation site by a combination of structural, biochemical, and spectroscopic approaches. Refinement of the PFL-AE crystal structure reveals Na + as the most likely ion present in the solved structures, and pulsed electron nuclear double resonance (ENDOR) demonstrates that the same cation site is occupied by 23 Na in the solution state of the as-isolated enzyme. A SAM carboxylate-oxygen is an M + ligand, and EPR and circular dichroism spectroscopies reveal that both the site occupancy and the identity of the cation perturb the electronic properties of the SAM-chelated iron-sulfur cluster. ENDOR studies of the PFL-AE/[ 13 C-methyl]-SAM complex show that the target sulfonium positioning varies with the cation, while the observation of an isotropic hyperfine coupling to the cation by ENDOR measurements establishes its intimate, SAM-mediated interaction with the cluster. This monovalent cation site controls enzyme activity: (i) PFL-AE in the absence of any simple monovalent cations has little-no activity; and (ii) among monocations, going down Group 1 of the periodic table from Li + to Cs + , PFL-AE activity sharply maximizes at K + , with NH 4 + closely matching the efficacy of K + . PFL-AE is thus a type I M + -activated enzyme whose M + controls reactivity by interactions with the cosubstrate, SAM, which is bound to the catalytic iron-sulfur cluster.

  7. One-step purification and characterization of alginate lyase from a clinical Pseudomonas aeruginosa with destructive activity on bacterial biofilm

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    Parinaz Ghadam

    2017-05-01

    Full Text Available Objective(s: Pseudomonas aeruginosais a Gram-negative and aerobic rod bacterium that displays mucoid and non-mucoid phenotype. Mucoid strains secrete alginate, which is the main agent of biofilms in chronic P. aeruginosa infections, show high resistance to antibiotics; consequently, the biological disruption of mucoid P. aeruginosa biofilms is an attractive area of study for researchers. Alginate lyase gene (algl is a member of alginate producing operon which by glycosidase activity produces primer for other enzymes in this cluster. Also this activity can destroy the extracellular alginate; therefore this enzyme participates in alginate production and destruction pathway. Alginate lyase causes detachment of a biofilm by reducing its adhesion to the surfaces, and increases phagocytosis and antibiotic susceptibility. In this study, alginate lyase was purified in just one step and its properties were investigated. Materials and Methods: The purification was done by affinity chromatography, analysed by SDS-PAGE, and its effect on P. aeruginosa biofilms was surveyed by micro titer plate assay and SEM. The substrate specificity of the enzyme was determined by PCR. Results: Alginate lyase from isolate 48 was purified in one step. It is more thermally resistant than alginate lyase from Pseudomonas aeruginosa PAO1 and poly M, poly G and poly MG alginate were the substrate of this enzyme. Moreover, it has an eradication effect on biofilms from P. aeruginosa 48 and PAO1. Conclusion: In this study an alginate lyase with many characteristics suitable in medicine such as thermal stability, effective on poly M alginate, and bacterial biofilm destructive was introduced and purified.

  8. Characterization of AlgMsp, an alginate lyase from Microbulbifer sp. 6532A.

    Directory of Open Access Journals (Sweden)

    Steven M Swift

    Full Text Available Alginate is a polysaccharide produced by certain seaweeds and bacteria that consists of mannuronic acid and guluronic acid residues. Seaweed alginate is used in food and industrial chemical processes, while the biosynthesis of bacterial alginate is associated with pathogenic Pseudomonas aeruginosa. Alginate lyases cleave this polysaccharide into short oligo-uronates and thus have the potential to be utilized for both industrial and medicinal applications. An alginate lyase gene, algMsp, from Microbulbifer sp. 6532A, was synthesized as an E.coli codon-optimized clone. The resulting 37 kDa recombinant protein, AlgMsp, was expressed, purified and characterized. The alginate lyase displayed highest activity at pH 8 and 0.2 M NaCl. Activity of the alginate lyase was greatest at 50°C; however the enzyme was not stable over time when incubated at 50°C. The alginate lyase was still highly active at 25°C and displayed little or no loss of activity after 24 hours at 25°C. The activity of AlgMsp was not dependent on the presence of divalent cations. Comparing activity of the lyase against polymannuronic acid and polyguluronic acid substrates showed a higher turnover rate for polymannuronic acid. However, AlgMSP exhibited greater catalytic efficiency with the polyguluronic acid substrate. Prolonged AlgMsp-mediated degradation of alginate produced dimer, trimer, tetramer, and pentamer oligo-uronates.

  9. Highly Active and Specific Tyrosine Ammonia-Lyases from Diverse Origins Enable Enhanced Production of Aromatic Compounds in Bacteria and Saccharomyces cerevisiae

    DEFF Research Database (Denmark)

    Jendresen, Christian Bille; Stahlhut, Steen Gustav; Li, Mingji

    2015-01-01

    Phenylalanine and tyrosine ammonia-lyases form cinnamic acid and p-coumaric acid, which are precursors of a wide range of aromatic compounds of biotechnological interest. Lack of highly active and specific tyrosine ammonia-lyases has previously been a limitation in metabolic engineering approaches...

  10. The replicative DNA polymerase of herpes simplex virus 1 exhibits apurinic/apyrimidinic and 5′-deoxyribose phosphate lyase activities

    Science.gov (United States)

    Bogani, Federica; Boehmer, Paul E.

    2008-01-01

    Base excision repair (BER) is essential for maintaining genome stability both to counter the accumulation of unusual bases and to protect from base loss in the DNA. Herpes simplex virus 1 (HSV-1) is a large dsDNA virus that encodes its own DNA replication machinery, including enzymes involved in nucleotide metabolism. We report on a replicative family B and a herpesvirus-encoded DNA Pol that possesses DNA lyase activity. We have discovered that the catalytic subunit of the HSV-1 DNA polymerase (Pol) (UL30) exhibits apurinic/apyrimidinic (AP) and 5′-deoxyribose phosphate (dRP) lyase activities. These activities are integral to BER and lead to DNA cleavage on the 3′ side of abasic sites and 5′-dRP residues that remain after cleavage by 5′-AP endonuclease. The UL30-catalyzed reaction occurs independently of divalent cation and proceeds via a Schiff base intermediate, indicating that it occurs via a lyase mechanism. Partial proteolysis of the Schiff base shows that the DNA lyase activity resides in the Pol domain of UL30. These observations together with the presence of a virus-encoded uracil DNA glycosylase indicates that HSV-1 has the capacity to perform critical steps in BER. These findings have implications on the role of BER in viral genome maintenance during lytic replication and reactivation from latency. PMID:18695225

  11. The effect of ultraviolet treatment on enzymatic activity and total phenolic content of minimally processed potato slices.

    Science.gov (United States)

    Teoh, Li Shing; Lasekan, Ola; Adzahan, Noranizan Mohd; Hashim, Norhashila

    2016-07-01

    In this work, potato slices were exposed to different doses of UV-C irradiation (i.e. 2.28, 6.84, 11.41, and 13.68 kJ m -2 ) with or without pretreatment [i.e. ascorbic acid and calcium chloride (AACCl) dip] and stored at 4 ± 1 °C. Changes in enzymatic activities of polyphenol oxidase (PPO), peroxidase (POD) and phenylalanine ammonia lyase (PAL), as well as total phenolic content (TPC) were investigated after 0, 3, 7 and 10 days of storage. Results showed that untreated and UV-C treated potato slices at 13.68 kJ m -2 dosage level showed significantly higher PPO, POD and PAL activities. Conversely, untreated potato slices showed the lowest TPC during storage period. Potato slices subjected to AACCl dip plus UV-C at 6.84 kJ m -2 produced lower PPO, POD and PAL activities, as well as maintained a high TPC during storage.

  12. Kynurenine aminotransferase III and glutamine transaminase L are identical enzymes that have cysteine S-conjugate β-lyase activity and can transaminate L-selenomethionine.

    Science.gov (United States)

    Pinto, John T; Krasnikov, Boris F; Alcutt, Steven; Jones, Melanie E; Dorai, Thambi; Villar, Maria T; Artigues, Antonio; Li, Jianyong; Cooper, Arthur J L

    2014-11-07

    Three of the four kynurenine aminotransferases (KAT I, II, and IV) that synthesize kynurenic acid, a neuromodulator, are identical to glutamine transaminase K (GTK), α-aminoadipate aminotransferase, and mitochondrial aspartate aminotransferase, respectively. GTK/KAT I and aspartate aminotransferase/KAT IV possess cysteine S-conjugate β-lyase activity. The gene for the former enzyme, GTK/KAT I, is listed in mammalian genome data banks as CCBL1 (cysteine conjugate beta-lyase 1). Also listed, despite the fact that no β-lyase activity has been assigned to the encoded protein in the genome data bank, is a CCBL2 (synonym KAT III). We show that human KAT III/CCBL2 possesses cysteine S-conjugate β-lyase activity, as does mouse KAT II. Thus, depending on the nature of the substrate, all four KATs possess cysteine S-conjugate β-lyase activity. These present studies show that KAT III and glutamine transaminase L are identical enzymes. This report also shows that KAT I, II, and III differ in their ability to transaminate methyl-L-selenocysteine (MSC) and L-selenomethionine (SM) to β-methylselenopyruvate (MSP) and α-ketomethylselenobutyrate, respectively. Previous studies have identified these seleno-α-keto acids as potent histone deacetylase inhibitors. Methylselenol (CH3SeH), also purported to have chemopreventive properties, is the γ-elimination product of SM and the β-elimination product of MSC catalyzed by cystathionine γ-lyase (γ-cystathionase). KAT I, II, and III, in part, can catalyze β-elimination reactions with MSC generating CH3SeH. Thus, the anticancer efficacy of MSC and SM will depend, in part, on the endogenous expression of various KAT enzymes and cystathionine γ-lyase present in target tissue coupled with the ability of cells to synthesize in situ either CH3SeH and/or seleno-keto acid metabolites. © 2014 by The American Society for Biochemistry and Molecular Biology, Inc.

  13. The hydroxynitrile lyase from almond: crystal structure and mechanistical studies

    International Nuclear Information System (INIS)

    Dreveny, Ingrid

    2001-09-01

    Cyanogenesis is a defense process of several thousand plant species. Hydroxynitrile lyase (HNL), a key enzyme of this process, cleaves a cyanohydrin precursor into hydrocyanic acid and the corresponding aldehyde or ketone. The reverse reaction constitutes an important tool in industrial biocatalysis. Different classes of hydroxynitrile lyases have convergently evolved from FAD-dependent oxidoreductases, α/β hydrolases and alcohol dehydrogenases. The FAD-dependent hydroxynitrile lyases (FAD-HNLs) carry a flavin cofactor whose redox properties appear to be unimportant for catalysis. The high resolution crystal structure of the hydroxynitrile lyase from almond (Prunus amygdalus), PaHNL1, has been determined and constitutes the first 3D structure of an FAD-HNL. The overall fold and the architecture of the active site region showed that PaHNL1 belongs to the glucose-methanol-choline-oxidoreductase family, with closest structural similarity to glucose oxidase. There is strong evidence from the sequence and the reaction product that FAD-dependent hydroxynitrile lyases have evolved from an aryl alcohol oxidizing precursor. Structures of PaHNL1 in complex with its natural substrate mandelonitrile and the competitive inhibitor benzyl alcohol provided insight into the residues involved in catalysis and a mechanism without participation of the cofactor could be suggested. Although the catalytic residues differ between the α/β-hydrolase-type HNLs and PaHNL1, common general features relevant for hydroxynitrile lyase activity could be proposed. (author)

  14. Production and Purification of a Novel Xanthan Lyase from a Xanthan-Degrading Microbacterium sp. Strain XT11

    Directory of Open Access Journals (Sweden)

    Fan Yang

    2014-01-01

    Full Text Available A xanthan lyase was produced and purified from the culture supernatant of an excellent xanthan-modifying strain Microbacterium sp. XT11. Xanthan lyase was induced by xanthan but was inhibited by its structural monomer glucose. Its production by strain XT11 is much higher than that by all other reported strains. The purified xanthan lyase has a molecular mass of 110 kDa and a specific activity of 28.2 U/mg that was much higher than that of both Paenibacillus and Bacillus lyases. It was specific on the pyruvated mannosyl residue in the intact xanthan molecule, but about 50% lyase activity remained when xanthan was partially depyruvated. Xanthan lyase was optimally active at pH 6.0–6.5 and 40°C and alkali-tolerant at a high pH value of 11.0. The metal ions including K+, Ca2+, Na+, Mg2+, Mn2+, and Li+ strongly stimulated xanthan lyase activity but ions Zn2+ and Cu2+ were its inhibitor. Xanthan lyase should be a novel enzyme different from the other xanthan lyases ever reported.

  15. New lupane triterpenoids from Solidago canadensis that inhibit the lyase activity of DNA polymerase beta.

    Science.gov (United States)

    Chaturvedula, V S Prakash; Zhou, Bing-Nan; Gao, Zhijie; Thomas, Shannon J; Hecht, Sidney M; Kingston, David G I

    2004-12-01

    Bioassay-directed fractionation of a methyl ethyl ketone extract of Solidago canadensis L. (Asteraceae), using an assay to detect the lyase activity of DNA polymerase beta, resulted in the isolation of the four new lupane triterpenoids 1-4 and the seven known compounds lupeol, lupeyl acetate, ursolic acid, cycloartenol, cycloartenyl palmitate, alpha-amyrin acetate, and stigmasterol. The structures of the new compounds were established as 3beta-(3R-acetoxyhexadecanoyloxy)-lup-20(29)-ene (1), 3beta-(3-ketohexadecanoyloxy)-lup-20(29)-ene (2), 3beta-(3R-acetoxyhexadecanoyloxy)-29-nor-lupan-20-one (3), and 3beta-(3-hetohexadecanoyloxy)-29-nor-lupan-20-one (4), respectively, on the basis of extensive 1D and 2D NMR spectroscopic interpretation and chemical modification studies. All 11 compounds were inhibitory to the lyase activity of DNA polymerase beta.

  16. Isolation and characterization of an Antarctic Flavobacterium strain with agarase and alginate lyase activities

    Directory of Open Access Journals (Sweden)

    Lavín Paris

    2016-09-01

    Full Text Available Several bacteria that are associated with macroalgae can use phycocolloids as a carbon source. Strain INACH002, isolated from decomposing Porphyra (Rhodophyta, in King George Island, Antarctica, was screened and characterized for the ability to produce agarase and alginate-lyase enzymatic activities. Our strain INACH002 was identified as a member of the genus Flavobacterium, closely related to Flavobacterium faecale, using 16S rRNA gene analysis. The INACH002 strain was characterized as psychrotrophic due to its optimal temperature (17ºC and maximum temperature (20°C of growth. Agarase and alginate-lyase displayed enzymatic activities within a range of 10°C to 50°C, with differences in the optimal temperature to hydrolyze agar (50°C, agarose (50°C and alginate (30°C during the first 30 min of activity. Strain Flavobacterium INACH002 is a promising Antarctic biotechnological resource; however, further research is required to illustrate the structural and functional bases of the enzymatic performance observed during the degradation of different substrates at different temperatures.

  17. Activity-Oriented Design of Health Pal: A Smart Phone for Elders' Healthcare Support

    Directory of Open Access Journals (Sweden)

    Fan Shih-Chen

    2008-01-01

    Full Text Available Wireless telephones and personal digital assistants are emerging, as the information hubs connect their human users with assorted electronic devices and the World Wide Web. As such, they quickly become the de facto basis for personalized information services. The Kannon project team at the National Chiao Tung University (NCTU in Taiwan is developing a ubiquitous service infrastructure for elders' healthcare support. Among their deliverables, there is a PDA Phone, christened Health Pal, which can communicate with Bluetooth/ZigBee devices, uni¬ver¬sal plug-and-play (UPnP e-home service platforms, and online healthcare providers to offer 24/7 healthcare services to elderly people. This paper presents the early results of this effort including the functional and operational concepts of Health Pal as well as the activity-oriented approach of its design. Preliminary results of its usefulness and usability evaluations are reported. A comparison of this platform against several similar prototypes was also included to illustrate the advantage of applying activity-oriented design approach to human-computer interactions.

  18. Activity-Oriented Design of Health Pal: A Smart Phone for Elders' Healthcare Support

    Directory of Open Access Journals (Sweden)

    Shih-Chen Fan

    2008-02-01

    Full Text Available Wireless telephones and personal digital assistants are emerging, as the information hubs connect their human users with assorted electronic devices and the World Wide Web. As such, they quickly become the de facto basis for personalized information services. The Kannon project team at the National Chiao Tung University (NCTU in Taiwan is developing a ubiquitous service infrastructure for elders' healthcare support. Among their deliverables, there is a PDA Phone, christened Health Pal, which can communicate with Bluetooth/ZigBee devices, uni¬ver¬sal plug-and-play (UPnP e-home service platforms, and online healthcare providers to offer 24/7 healthcare services to elderly people. This paper presents the early results of this effort including the functional and operational concepts of Health Pal as well as the activity-oriented approach of its design. Preliminary results of its usefulness and usability evaluations are reported. A comparison of this platform against several similar prototypes was also included to illustrate the advantage of applying activity-oriented design approach to human-computer interactions.

  19. Wound-induced ethylene synthesis and expression and formation of 1-aminocyclopropane-1-carboxylate (ACC) synthase, ACC oxidase, phenylalanine ammonia-lyase, and peroxidase in wounded mesocarp tissue of Cucurbita maxima.

    Science.gov (United States)

    Kato, M; Hayakawa, Y; Hyodo, H; Ikoma, Y; Yano, M

    2000-04-01

    1-Aminocyclopropane-1-carboxylate (ACC) synthase was rapidly induced in mesocarp tissue of Cucurbita maxima after wounding in the cut surface layer in 1 mm thickness (ca. 9 cells) (first layer) in both the enzyme activity and the levels of transcript. This led to a rapid accumulation of ACC and hence ethylene production. In the inside tissue (1-2 mm) (second layer), no significant induction of ACC synthase was observed, which resulted in a low level of ACC, although ethylene was evolved at a much lower rate than the first one. In contrast to ACC synthase, ACC oxidase was induced markedly in both the first and second layers and the development of its activity and the levels of mRNA remained high until later stages. It was considered that wound ethylene was closely associated with the development of ACC oxidase, since 2,5-norbornadiene (NBD), an inhibitor of ethylene action, substantially suppressed it. Phenylalanine ammonia-lyase (PAL) greatly increased in activity after wounding similarly to that of ACC synthase, in which increase in PAL activity occurred predominantly in the first layer. Induction of peroxidase activity after wounding had a close correlation in profile with that of ACC oxidase in that marked increases in the activity were observed in both the first and second layers and were strongly suppressed by NBD application. Four peroxidase isozymes were found by PAGE, among which a fraction was newly detected after wounding.

  20. Proliferation related acidic leucine-rich protein PAL31 functions as a caspase-3 inhibitor

    International Nuclear Information System (INIS)

    Sun Weiyong; Kimura, Hiromichi; Hattori, Naka; Tanaka, Satoshi; Matsuyama, Shigemi; Shiota, Kunio

    2006-01-01

    Proliferation related acidic leucine-rich protein PAL31 (PAL31) is expressed in proliferating cells and consists of 272 amino acids with a tandem structure of leucine-rich repeats in the N-terminus and a highly acidic region with a putative nuclear localization signal in the C-terminus. We previously reported that PAL31 is required for cell cycle progression. In the present study, we found that the antisense oligonucleotide of PAL31 induced apoptosis to the transfected Nb2 cells. Stable transfectants, in which PAL31 was regulated by an inducible promoter, were generated to gain further insight into the signaling role of PAL31 in the regulation of apoptosis. Expression of PAL31 resulted in the marked rescue of Rat1 cells from etoposide and UV radiation-induced apoptosis and the cytoprotection was correlated with the levels of PAL31 protein. Thus, cytoprotection from apoptosis is a physiological function of PAL31. PAL31 can suppress caspase-3 activity but not cytochrome c release in vitro, indicating that PAL31 is a direct caspase-3 inhibitor. In conclusion, PAL31 is a multifunctional protein working as a cell cycle progression factor as well as a cell survival factor

  1. [Effect of exogenous sucrose on growth and active ingredient content of licorice seedlings under salt stress conditions].

    Science.gov (United States)

    Liu, Fu-zhi; Yang, Jun

    2015-11-01

    Licorice seedlings were taken as experimental materials, an experiment was conducted to study the effects of exogenous sucrose on growth and active ingredient content of licorice seedlings under NaCl stress conditions. The results of this study showed that under salt stress conditions, after adding a certain concentration of exogenous sucrose, the licorice seedlings day of relative growth rate was increasing, and this stress can be a significant weakening effect, indicating that exogenous sucrose salt stress-relieving effect. The total flavonoids and phenylalanine ammonia lyase (PAL) activity were significantly increased, the exogenous sucrose can mitigated the seedling roots under salt stress, the licorice flavonoid content in the enhanced growth was largely due to the activity of PAL an increased, when the concentration of exogenous sucrose wae 10 mmol x L(-1), PAL activity reaching a maximum, when the concentration of exogenous sucrose was 15 mmol x L(-1), PAL activity turned into a downward trend, the results indicating that this mitigation has concentration effect. After applying different concentrations of exogenous sugar, the contents of liquiritin changes with the change of flavonoids content was similar. After applying different concentrations of exogenous sucrose, the content of licorice acid under salt stress was higher than the levels were not reached during salt stress, the impact of exogenous sucrose concentration gradient of licorice acid accumulation was not obvious.

  2. Phenolics and Flavonoids Compounds, Phenylanine Ammonia Lyase and Antioxidant Activity Responses to Elevated CO2 in Labisia pumila (Myrisinaceae

    Directory of Open Access Journals (Sweden)

    Hawa Z.E. Jaafar

    2012-05-01

    Full Text Available A split plot 3 × 3 experiment was designed to examine the impact of three concentrations of CO2 (400, 800 and 1,200 µmol·mol−1 on the phenolic and flavonoid compound profiles, phenylalanine ammonia lyase (PAL and antioxidant activity in three varieties of Labisia pumila Benth. (var. alata, pumila and lanceolata after 15 weeks of exposure. HPLC analysis revealed a strong influence of increased CO2 concentration on the modification of phenolic and flavonoid profiles, whose intensity depended on the interaction between CO2 levels and L. pumila varieties. Gallic acid and quercetin were the most abundant phenolics and flavonoids commonly present in all the varieties. With elevated CO2 (1,200 µmol·mol−1 exposure, gallic acid increased tremendously, especially in var. alata and pumila (101–111%, whilst a large quercetin increase was noted in var. lanceolata (260%, followed closely by alata (201%. Kaempferol, although detected under ambient CO2 conditions, was undetected in all varieties after exposure. Instead, caffeic acid was enhanced tremendously in var. alata (338~1,100% and pumila (298~433%. Meanwhile, pyragallol and rutin were only seen in var. alata (810 µg·g−1 DW and pumila (25 µg·g−1 DW, respectively, under ambient conditions; but the former compound went undetected in all varieties while rutin continued to increase by 262% after CO2 enrichment. Interestingly, naringenin that was present in all varieties under ambient conditions went undetected under enrichment, except for var. pumila where it was enhanced by 1,100%. PAL activity, DPPH and FRAP also increased with increasing CO2 levels implying the possible improvement of health-promoting quality of Malaysian L. pumila

  3. Utilization of Aspergillus oryzae to produce pectin lyase from various agro-industrial residues

    OpenAIRE

    Koser, Safia; Anwar, Zahid; Iqbal, Zafar; Anjum, Awais; Aqil, Tahir; Mehmood, Sajid; Irshad, Muhammad

    2014-01-01

    The present study was aimed to investigate the culture influence on pectin lyase production potential of fungal strain Aspergillus oryzae. The enzyme profile of A. oryzae showed highest activity of pectin lyase after 3rd day of incubation on lemon peel waste under solid state fermentation conditions. To induce the pectin lyase synthesis capability of A. oryzae at optimal level various culture variables including physical and nutritional parameters were optimized by adopting classical optimiza...

  4. Alfalfa contains substantial 9-hydroperoxide lyase activity and a 3Z:2E-enal isomerase

    NARCIS (Netherlands)

    Vliegenthart, J.F.G.; Noordermeer, M.A.; Veldink, G.A.

    1999-01-01

    Fatty acid hydroperoxides formed by lipoxygenase can be cleaved by hydroperoxide lyase resulting in the formation of short-chain aldehydes and omega-oxo acids. Plant hydroperoxide lyases use 13- or 9-hydroperoxy linoleic and linolenic acid as substrates. Alfalfa (Medicago sativa L.) has been

  5. [Construction and high-density fermentation of alkaline pectate lyase high-yield yeast].

    Science.gov (United States)

    Wang, Xiaowen; Xiang, La; Xu, Ting; Lu, Zhenghui; Zhang, Guimin

    2017-12-25

    Pectate lyase is widely applied in ramie degumming and fabric bioscouring in the textile industry. Compared to conventional processes that involve high alkaline and high temperature treatment, enzyme based treatments have significant advantages in fibers protectiveness, improved efficiency of refining, reduced energy consumption and pollution. Hence, it would be highly desirable to construct high-yield alkaline pectate lyase engineered strains and reduce the pectate lyase production cost. In the previous study, pectate lyase gene pel from Bacillus subtilis168 was expressed in Pichia pastoris GS115 after codon usage optimization based on the vector pHBM905A. To improve the expression level, the vector pHBM905BDM with optimized promoter and signal peptide was used to express the optimized gene pels in GS115. The transformant had increased activity from 68 U/mL to 100 U/mL with the improvement in the transcription level by 27% measured by qPCR. The transformants were further screened on pectin plates, where higher halo forming strains were picked for shake-flask fermentation and strain GS115-pHBM905BDM-pels4 showed the highest activity of 536 U/mL. Then plasmid pPIC9K-pels was constructed and electroporated into the GS115-pHBM905BDM-pels4 cells. Subsequently, high-copy transformant was screened by using the medium containing antibiotics G418, strain GS115-pHBM905BDMpPIC9K- pels1 was identified with increased activity of 770 U/mL and the copy number of pels was 7 confirmed by qPCR. Finally, the activity of pectate lyase produced by GS115-pHBM905BDM-pPIC9K-pels1reached to 2 271 U/mL in a 5-L fermentor. The activity of pectate lyase in our study reached the highest level of expression in P. pastoris, showing good application potential in the textile industry.

  6. Becoming an Effective Peer Assisted Learning (PAL) Leader

    Science.gov (United States)

    West, Harry; Jenkins, Rhiannon; Hill, Jennifer

    2017-01-01

    This Directions paper, written by two former Peer Assisted Learning (PAL) Leaders, (West and Jenkins) and a PAL Tutor (Hill), will support any geographer considering a PAL role. It reflects their experiences of participating in a PAL scheme at the University of the West of England (UWE), Bristol, United Kingdom, and research conducted with…

  7. Active Detectors for Plasma Soft X-Ray Detection at PALS

    Directory of Open Access Journals (Sweden)

    C. Granja

    2010-01-01

    Full Text Available This paper summarizes the work carried out for an experimental study of low-energy nuclear excitation by laser-produced plasma at the PALS Prague laser facility. We describe the adaptation and shielding of single-quantum active radiation detectors developed at IEAP CTU Prague to facilitate their operation inside the laser interaction chamber in the vicinity of the plasma target. The goal of this effort is direct real-time single-quantum detection of plasma soft X-ray radiation with energy above a few keV and subsequent identification of the decay of the excited nuclear states via low-energy gamma rays in a highly radiative environment with strong electromagnetic interference.

  8. Xylella fastidiosa esterase rather than hydroxynitrile lyase.

    Science.gov (United States)

    Torrelo, Guzman; Ribeiro de Souza, Fayene Zeferino; Carrilho, Emanuel; Hanefeld, Ulf

    2015-03-02

    In 2009, we reported that the product of the gene SCJ21.16 (XFa0032) from Xylella fastidiosa, a xylem-restricted plant pathogen that causes a range of diseases in several important crops, encodes a protein (XfHNL) with putative hydroxynitrile lyase activity. Sequence analysis and activity tests indicated that XfHNL exhibits an α/β-hydrolase fold and could be classified as a member of the family of FAD-independent HNLs. Here we provide a more detailed sequence analysis and new experimental data. Using pure heterologously expressed XfHNL we show that this enzyme cannot catalyse the cleavage/synthesis of mandelonitrile and that this protein is in fact a non-enantioselective esterase. Homology modelling and ligand docking simulations were used to study the active site and support these results. This finding could help elucidate the common ancestor of esterases and hydroxynitrile lyases with an α/β -hydrolase fold. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  9. Utilization of Aspergillus oryzae to produce pectin lyase from various agro-industrial residues

    Directory of Open Access Journals (Sweden)

    Safia Koser

    2014-07-01

    Full Text Available The present study was aimed to investigate the culture influence on pectin lyase production potential of fungal strain Aspergillus oryzae. The enzyme profile of A. oryzae showed highest activity of pectin lyase after 3rd day of incubation on lemon peel waste under solid state fermentation conditions. To induce the pectin lyase synthesis capability of A. oryzae at optimal level various culture variables including physical and nutritional parameters were optimized by adopting classical optimization technique. Therefore, through fermentation process optimization the production of pectin lyase was substantially induced up to the level of 875 U/mL, when fermentation medium of lemon peel waste inoculated with 5 mL spore suspension of A. oryzae. The optimal fermentation conditions for maximum pectin lyase yield were as: optimum pH 5, 70% moisture level and incubated at 40 °C in addition with 1% sterile glucose solution as readily available carbon source and 0.2% yeast extract as an inexpensive nitrogen supplement (1%. The results obtained in current investigation so far demonstrated that culture conditions have great influence on the pectin lyase production potential of A. oryzae.

  10. Characterization of a Novel Alginate Lyase from Marine Bacterium Vibrio furnissii H1

    Directory of Open Access Journals (Sweden)

    Xiaoyan Zhu

    2018-01-01

    Full Text Available Alginate lyases show great potential for industrial and medicinal applications, especially as an attractive biocatalyst for the production of oligosaccharides with special bioactivities. A novel alginate lyase, AlyH1, from the marine bacterium Vibrio furnissii H1, which has been newly isolated from rotten seaweed, was purified and characterized. The purified enzyme showed the specific activity of 2.40 U/mg. Its molecular mass was 35.8 kDa. The optimal temperature and pH were 40 °C and pH 7.5, respectively. AlyH1 maintained stability at neutral pH (7.0–8.0 and temperatures below 30 °C. Metal ions Na+, Mg2+, and K+ increased the activity of the enzyme. With sodium alginate as the substrate, the Km and Vmax values of AlyH1 were 2.28 mg/mL and 2.81 U/mg, respectively. AlyH1 exhibited activities towards both polyguluronate and polymannuronate, and preferentially degraded polyguluronate. Products prepared from sodium alginate by AlyH1 were displayed to be di-, tri-, and tetra-alginate oligosaccharides. A partial amino acid sequence (190 aa of AlyH1 analysis suggested that AlyH1 was an alginate lyase of polysaccharide lyase family 7. The sequence showed less than 77% identity to the reported alginate lyases. These data demonstrated that AlyH1 could be as a novel and potential candidate in application of alginate oligosaccharides production with low polymerization degrees.

  11. Effects of germination on the activities of amylases and phenolic enzymes in sorghum varieties grouped according to food end-use properties

    NARCIS (Netherlands)

    Dicko, M.H.; Gruppen, H.; Zouzouho, O.C.; Traore, A.S.; Berkel, van W.J.H.; Voragen, A.G.J.

    2006-01-01

    Fifty sorghum varieties were screened to determine the effects of germination on levels of starch, -amylase, -amylase, phenylalanine ammonia lyase (PAL), peroxidase (POX) and polyphenol oxidase (PPO). Germination decreased starch content, with amylose being more degraded than amylopectin. In

  12. Antioxidant characterization and sensory evaluation during storage of ultraviolet-B light exposed baby carrots (abstract)

    Science.gov (United States)

    Baby carrot processing induces wounding stress activation of phenylalanine ammonia-lyase (PAL), enhancing its nutrient content by increasing synthesis of secondary metabolites. Ultraviolet-B (UV-B) exposure further promotes the formation of soluble phenolic compounds, significantly increasing antiox...

  13. Molecular cloning, purification, and characterization of a novel polyMG-specific alginate lyase responsible for alginate MG block degradation in Stenotrophomas maltophilia KJ-2

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Su In; Kim, Hee Sook [Kyungsung Univ., Busan (Korea, Republic of). Dept. of Food Science and Biotechnology; Choi, Sung Hee; Lee, Eun Yeol [Kyung Hee Univ., Gyeonggi-do (Korea, Republic of). Dept. of Chemical Engineering

    2012-09-15

    A gene for a polyMG-specific alginate lyase possessing a novel structure was identified and cloned from Stenotrophomas maltophilia KJ-2 by using PCR with homologous nucleotide sequences-based primers. The recombinant alginate lyase consisting of 475 amino acids was purified on Ni-Sepharose column and exhibited the highest activity at pH 8 and 40 C. Interestingly, the recombinant alginate lyase was expected to have a similar catalytic active site of chondroitin B lyase but did not show chondroitin lyase activity. In the test of substrate specificity, the recombinant alginate lyase preferentially degraded the glycosidic bond of polyMG-block than polyM-block and polyG-block. The chemical structures of the degraded alginate oligosaccharides were elucidated to have mannuronate (M) at the reducing end on the basis of NMR analysis, supporting that KJ-2 polyMG-specific alginate lyase preferably degraded the glycosidic bond in M-G linkage than that in G-M linkage. The KJ-2 polyMG-specific alginate lyase can be used in combination with other alginate lyases for a synergistic saccharification of alginate. (orig.)

  14. Evaluation of the hydroxynitrile lyase activity in cell cultures of capulin (Prunus serotina).

    Science.gov (United States)

    Hernández, Liliana; Luna, Héctor; Navarro-Ocaña, Arturo; Olivera-Flores, Ma Teresa de Jesús; Ayala, Ivon

    2008-07-01

    Enzymatic preparations obtained from young plants and cell cultures of capulin were screened for hydroxynitrile lyase activity. The three week old plants, grown under sterile conditions, were used to establish a solid cell culture. Crude preparations obtained from this plant material were evaluated for the transformation of benzaldehyde to the corresponding cyanohydrin (mandelonitrile). The results show that the crude material from roots, stalks, and leaves of young plants and calli of roots, stalks, internodes and petioles biocatalyzed the addition of hydrogen cyanide (HCN) to benzaldehyde with a modest to excellent enantioselectivity.

  15. Impact of different alginate lyases on combined cellulase–lyase saccharification of brown seaweed

    DEFF Research Database (Denmark)

    Manns, Dirk Martin; Nyffenegger, Christian; Saake, B.

    2016-01-01

    -guluronic acid. When applied together with a fungal cellulase preparation (Cellic®CTec2) at pH 6 and 40 °C on a glucan rich brown seaweed Laminaria digitata the viscosity decreased in the initial minutes while measurable alginate degradation occurred primarily within the first 1–2 hours of reaction. Whereas FALy......, indicating that the degradation of mannuronic acid blocks inhibited cellulase catalyzed glucose release from L. digitata. Nevertheless, combined alginate lyase and cellulase treatment for 24 hours released all potential glucose regardless of the applied lyase. The enzymatic treatment moreover induced...

  16. Enhancing RGI lyase thermostability by targeted single point mutations

    DEFF Research Database (Denmark)

    Silva, Inês R.; Larsen, Dorte Møller; Jers, Carsten

    2013-01-01

    Rhamnogalacturonan I lyase (RGI lyase) (EC 4.2.2.-) catalyzes the cleavage of rhamnogalacturonan I in pectins by β-elimination. In this study the thermal stability of a RGI lyase (PL 11) originating from Bacillus licheniformis DSM 13/ATCC14580 was increased by a targeted protein engineering...

  17. Internal consistency, concurrent validity, and discriminant validity of a measure of public support for policies for active living in transportation (PAL-T) in a population-based sample of adults.

    Science.gov (United States)

    Fuller, Daniel; Gauvin, Lise; Fournier, Michel; Kestens, Yan; Daniel, Mark; Morency, Patrick; Drouin, Louis

    2012-04-01

    Active living is a broad conceptualization of physical activity that incorporates domains of exercise; recreational, household, and occupational activities; and active transportation. Policy makers develop and implement a variety of transportation policies that can influence choices about how to travel from one location to another. In making such decisions, policy makers act in part in response to public opinion or support for proposed policies. Measures of the public's support for policies aimed at promoting active transportation can inform researchers and policy makers. This study examined the internal consistency, and concurrent and discriminant validity of a newly developed measure of the public's support for policies for active living in transportation (PAL-T). A series of 17 items representing potential policies for promoting active transportation was generated. Two samples of participants (n = 2,001 and n = 2,502) from Montreal, Canada, were recruited via random digit dialling. Analyses were conducted on the combined data set (n = 4,503). Participants were aged 18 through 94 years (58% female). The concurrent and discriminant validity of the PAL-T was assessed by examining relationships with physical activity and smoking. To explore the usability of the PAL-T, predicted scale scores were compared to the summed values of responses. Results showed that the internal consistency of the PAL-T was 0.70. Multilevel regression demonstrated no relationship between the PAL-T and smoking status (p > 0.05) but significant relationships with utilitarian walking (p public opinion can inform policy makers and support advocacy efforts aimed at making built environments more suitable for active transportation while allowing researchers to examine the antecedents and consequences of public support for policies.

  18. PAL: an object-oriented programming library for molecular evolution and phylogenetics.

    Science.gov (United States)

    Drummond, A; Strimmer, K

    2001-07-01

    Phylogenetic Analysis Library (PAL) is a collection of Java classes for use in molecular evolution and phylogenetics. PAL provides a modular environment for the rapid construction of both special-purpose and general analysis programs. PAL version 1.1 consists of 145 public classes or interfaces in 13 packages, including classes for models of character evolution, maximum-likelihood estimation, and the coalescent, with a total of more than 27000 lines of code. The PAL project is set up as a collaborative project to facilitate contributions from other researchers. AVAILIABILTY: The program is free and is available at http://www.pal-project.org. It requires Java 1.1 or later. PAL is licensed under the GNU General Public License.

  19. A radiometric technique for the measurement of adenylosuccinate lyase

    International Nuclear Information System (INIS)

    Park, K.W.; Tyagi, A.K.; Cooney, D.A.

    1980-01-01

    When radioactive adenylosuccinic acid (AMP-S) is metabolized to AMP and fumaric acid by the enzyme adenylosuccinate lyase (EC 4.3.2.2), a proton is released to the solvent as 3 H 2 O. This removal is believed to be stereospecifically identical to that catalyzed by the enzyme, L-aspartase, and therefore entails the loss of a proton from C-3 of the dicarboxylic acid moiety of the nucleotide. Advantage has been taken of this fact in the design of a facile assay for this enzyme. The assay permits the simultaneous estimation of the lyase activity in a large battery of samples; it is not interfered with by opalescent or proteinaceous suspensions; it is accurate and outstandingly sensitive. (Auth.)

  20. Allyl isothiocyanate enhances shelf life of minimally processed shredded cabbage.

    Science.gov (United States)

    Banerjee, Aparajita; Penna, Suprasanna; Variyar, Prasad S

    2015-09-15

    The effect of allyl isothiocyanate (AITC), in combination with low temperature (10°C) storage on post harvest quality of minimally processed shredded cabbage was investigated. An optimum concentration of 0.05μL/mL AITC was found to be effective in maintaining the microbial and sensory quality of the product for a period of 12days. Inhibition of browning was shown to result from a down-regulation (1.4-fold) of phenylalanine ammonia lyase (PAL) gene expression and a consequent decrease in PAL enzyme activity and o-quinone content. In the untreated control samples, PAL activity increased following up-regulation in PAL gene expression that could be linearly correlated with enhanced o-quinone formation and browning. The efficacy of AITC in extending the shelf life of minimally processed shredded cabbage and its role in down-regulation of PAL gene expression resulting in browning inhibition in the product is reported here for the first time. Copyright © 2015 Elsevier Ltd. All rights reserved.

  1. In Silico Characterization of Pectate Lyase Protein Sequences from Different Source Organisms

    Directory of Open Access Journals (Sweden)

    Amit Kumar Dubey

    2010-01-01

    Full Text Available A total of 121 protein sequences of pectate lyases were subjected to homology search, multiple sequence alignment, phylogenetic tree construction, and motif analysis. The phylogenetic tree constructed revealed different clusters based on different source organisms representing bacterial, fungal, plant, and nematode pectate lyases. The multiple accessions of bacterial, fungal, nematode, and plant pectate lyase protein sequences were placed closely revealing a sequence level similarity. The multiple sequence alignment of these pectate lyase protein sequences from different source organisms showed conserved regions at different stretches with maximum homology from amino acid residues 439–467, 715–816, and 829–910 which could be used for designing degenerate primers or probes specific for pectate lyases. The motif analysis revealed a conserved Pec_Lyase_C domain uniformly observed in all pectate lyases irrespective of variable sources suggesting its possible role in structural and enzymatic functions.

  2. Crystallization and preliminary X-ray analysis of an exotype alginate lyase Atu3025 from Agrobacterium tumefaciens strain C58, a member of polysaccharide lyase family 15

    International Nuclear Information System (INIS)

    Ochiai, Akihito; Yamasaki, Masayuki; Mikami, Bunzo; Hashimoto, Wataru; Murata, Kousaku

    2006-01-01

    The crystallization and preliminary X-ray characterization of a family PL-15 exotype alginate lyase are presented. Almost all alginate lyases depolymerize alginate in an endolytical fashion via a β-elimination reaction. The alginate lyase Atu3025 from Agrobacterium tumefaciens strain C58, consisting of 776 amino-acid residues, is a novel exotype alginate lyase classified into polysaccharide lyase family 15. The enzyme was crystallized at 293 K by sitting-drop vapour diffusion with polyethylene glycol 4000 as a precipitant. Preliminary X-ray analysis showed that the Atu3025 crystal belonged to space group P2 1 and diffracted to 2.8 Å resolution, with unit-cell parameters a = 107.7, b = 108.3, c = 149.5 Å, β = 91.5°

  3. Characterization of C-S lyase from Lactobacillus delbrueckii subsp. bulgaricus ATCC BAA-365 and its potential role in food flavour applications.

    Science.gov (United States)

    Allegrini, Alessandra; Astegno, Alessandra; La Verde, Valentina; Dominici, Paola

    2017-04-01

    Volatile thiols have substantial impact on the aroma of many beverages and foods. Thus, the control of their formation, which has been linked to C-S lyase enzymatic activities, is of great significance in industrial applications involving food flavours. Herein, we have carried out a spectroscopic and functional characterization of a putative pyridoxal 5'-phosphate (PLP)-dependent C-S lyase from the lactic acid bacterium Lactobacillus delbrueckii subsp. bulgaricus ATCC BAA-365 (LDB C-S lyase). Recombinant LDB C-S lyase exists as a tetramer in solution and shows spectral properties of enzymes containing PLP as cofactor. The enzyme has a broad substrate specificity toward sulphur-containing amino acids with aminoethyl-L-cysteine and L-cystine being the most effective substrates over L-cysteine and L-cystathionine. Notably, the protein also reveals cysteine-S-conjugate β-lyase activity in vitro, and is able to cleave a cysteinylated substrate precursor into the corresponding flavour-contributing thiol, with a catalytic efficiency higher than L-cystathionine. Contrary to similar enzymes of other lactic acid bacteria however, LDB C-S lyase is not capable of α,γ-elimination activity towards L-methionine to produce methanethiol, which is a significant compound in flavour development. Based on our results, future developments can be expected regarding the flavour-forming potential of Lactobacillus C-S lyase and its use in enhancing food flavours. © The Authors 2017. Published by Oxford University Press on behalf of the Japanese Biochemical Society. All rights reserved.

  4. Kinetic and thermodynamic properties of alginate lyase and cellulase co-produced by Exiguobacterium species Alg-S5.

    Science.gov (United States)

    Mohapatra, Bidyut R

    2017-05-01

    In an effort to screen out the alginolytic and cellulolytic bacteria from the putrefying invasive seaweed Sargassum species accumulated off Barbados' coast, a potent bacterial strain was isolated. This bacterium, which simultaneously produced alginate lyase and cellulase, was identified as Exiguobacterium sp. Alg-S5 via the phylogenetic approach targeting the 16S rRNA gene. The co-produced alginate lyase and cellulase exhibited maximal enzymatic activity at pH 7.5 and at 40°C and 45°C, respectively. The K m and V max values recorded as 0.91mg/mL and 21.8U/mg-protein, respectively, for alginate lyase, and 10.9mg/mL and 74.6U/mg-protein, respectively, for cellulase. First order kinetic analysis of the thermal denaturation of the co-produced alginate lyase and cellulase in the temperature range from 40°C to 55°C revealed that both the enzymes were thermodynamically efficient by displaying higher activation energy and enthalpy of denaturation. These enzymatic properties indicate the potential industrial importance of this bacterium in algal biomass conversion. This appears to be the first report on assessing the efficacy of a bacterium for the co-production of alginate lyase and cellulase. Copyright © 2017 Elsevier B.V. All rights reserved.

  5. Ultrasound treatment on phenolic metabolism and antioxidant capacity of fresh-cut pineapple during cold storage.

    Science.gov (United States)

    Yeoh, Wei Keat; Ali, Asgar

    2017-02-01

    Ultrasound treatment at different power output (0, 25 and 29W) and exposure time (10 and 15min) was used to investigate its effect on the phenolic metabolism enzymes, total phenolic content and antioxidant capacity of fresh-cut pineapple. Following ultrasound treatment at 25 and 29W, the activity of phenylalanine ammonia lyase (PAL) was increased significantly (Ppineapple was significantly (Ppineapple. Results suggest that hormetic dosage of ultrasound treatment can enhance the activity of PAL and total phenolic content and hence the total antioxidant capacity to encounter with oxidative stress. Copyright © 2016. Published by Elsevier Ltd.

  6. A study of porosity of synthetic polymer nanoparticles using PALS

    Energy Technology Data Exchange (ETDEWEB)

    Pham, B; Smith, S V [Centre for Antimatter-Matter Studies, Australian Nuclear Science and Technology Organisation (ANSTO) NSW 2232 (Australia); Guagliardo, P; Williams, J; Samarin, S, E-mail: binh.pham@ansto.gov.au, E-mail: svs@ansto.gov.au [Centre for Antimatter-Matter Studies, School of Physics, University of Western Australia, WA 6009 (Australia)

    2011-01-01

    Positron annihilation lifetime spectroscopy (PALS) has been used to study the free volume in dry synthetic polymer nanoparticles of various sizes. A series of poly(styrene/divinyl benzene) particles with diameters in the range of 100 to 500 nm were synthesized and then carefully chemically treated using the sulfonation process, to increase their porosity. The particles were characterised by Scanning Electron Microscopy (SEM), light scattering and PALS. Light scattering gave larger size for the treated particles, reflecting the hydration effect and therefore the increase in porosity. PALS spectra of untreated and treated particles gave four and three life-time components, respectively. Analysis by PAScual version 1.3.0 program indicated there was a reduction in the intensity and the type of the micropores in the treated particles. The data suggest PALS is a sensitive tool for detecting changes in microporosity in particles. The conflicting results obtained for light scattering compared to PALS for chemically treated particles is difficult to resolve and suggests sample preparation of polymeric materials for PALS is the critical factor.

  7. Cloning and expression of isocitrate lyase from human round worm Strongyloides stercoralis

    Directory of Open Access Journals (Sweden)

    Siddiqui A.A.

    2000-09-01

    Full Text Available A full length cDNA (1463 bp encoding isocitrate lyase (EC 4.1.3.1 of Strongyloides stercoralis is described. The nucleotide sequence of this insert identified a cDNA coding for the isocitrate lyase. The conceptually translated amino acid sequence of the open reading frame for S. stercoralis isocitrate lyase encodes a 450 amino acid residue protein with an apparent molecular weight of 50 kDa and a predicted pl of 6.39. The sequence is 69 % A/T, reflecting a characteristic A/T codon bias of S. stercoralis. The amino acid sequence of S. stercoralis isocitrate lyase is compared with bifunctional glyoxylate cycle protein of Caenorhabditis elegans and isocitrate lyases from Chlamydomonas reinhardtii and Myxococcus xanthus. The full length cDNA of S. stercoralis was expressed in pRSET vector and bacteriophage T7 promoter based expression system. S. stercoralis lyase recombinant protein, purified via immobilized metal affinity chromatography, showed a molecular mass of 50 kDa on polyacrylamide gels. The role of isocitrate lyase in the glyoxylate cycle and energy metabolism of S. stercoralis is also discussed.

  8. Structural Snapshots of an Engineered Cystathionine-γ-lyase Reveal the Critical Role of Electrostatic Interactions in the Active Site

    Energy Technology Data Exchange (ETDEWEB)

    Yan, Wupeng; Stone, Everett; Zhang, Yan Jessie

    2017-02-01

    Enzyme therapeutics that can degrade l-methionine (l-Met) are of great interest as numerous malignancies are exquisitely sensitive to l-Met depletion. To exhaust the pool of methionine in human serum, we previously engineered an l-Met-degrading enzyme based on the human cystathionine-γ-lyase scaffold (hCGL-NLV) to circumvent immunogenicity and stability issues observed in the preclinical application of bacterially derived methionine-γ-lyases. To gain further insights into the structure–activity relationships governing the chemistry of the hCGL-NLV lead molecule, we undertook a biophysical characterization campaign that captured crystal structures (2.2 Å) of hCGL-NLV with distinct reaction intermediates, including internal aldimine, substrate-bound, gem-diamine, and external aldimine forms. Curiously, an alternate form of hCGL-NLV that crystallized under higher-salt conditions revealed a locally unfolded active site, correlating with inhibition of activity as a function of ionic strength. Subsequent mutational and kinetic experiments pinpointed that a salt bridge between the phosphate of the essential cofactor pyridoxal 5'-phosphate (PLP) and residue R62 plays an important role in catalyzing β- and γ-eliminations. Our study suggests that solvent ions such as NaCl disrupt electrostatic interactions between R62 and PLP, decreasing catalytic efficiency.

  9. Biochemical, Kinetic, and Spectroscopic Characterization of Ruegeria pomeroyi DddW--A Mononuclear Iron-Dependent DMSP Lyase.

    Directory of Open Access Journals (Sweden)

    Adam E Brummett

    Full Text Available The osmolyte dimethylsulfoniopropionate (DMSP is a key nutrient in marine environments and its catabolism by bacteria through enzymes known as DMSP lyases generates dimethylsulfide (DMS, a gas of importance in climate regulation, the sulfur cycle, and signaling to higher organisms. Despite the environmental significance of DMSP lyases, little is known about how they function at the mechanistic level. In this study we biochemically characterize DddW, a DMSP lyase from the model roseobacter Ruegeria pomeroyi DSS-3. DddW is a 16.9 kDa enzyme that contains a C-terminal cupin domain and liberates acrylate, a proton, and DMS from the DMSP substrate. Our studies show that as-purified DddW is a metalloenzyme, like the DddQ and DddP DMSP lyases, but contains an iron cofactor. The metal cofactor is essential for DddW DMSP lyase activity since addition of the metal chelator EDTA abolishes its enzymatic activity, as do substitution mutations of key metal-binding residues in the cupin motif (His81, His83, Glu87, and His121. Measurements of metal binding affinity and catalytic activity indicate that Fe(II is most likely the preferred catalytic metal ion with a nanomolar binding affinity. Stoichiometry studies suggest DddW requires one Fe(II per monomer. Electronic absorption and electron paramagnetic resonance (EPR studies show an interaction between NO and Fe(II-DddW, with NO binding to the EPR silent Fe(II site giving rise to an EPR active species (g = 4.29, 3.95, 2.00. The change in the rhombicity of the EPR signal is observed in the presence of DMSP, indicating that substrate binds to the iron site without displacing bound NO. This work provides insight into the mechanism of DMSP cleavage catalyzed by DddW.

  10. Hematopoietic sphingosine 1-phosphate lyase deficiency decreases atherosclerotic lesion development in LDL-receptor deficient mice.

    Directory of Open Access Journals (Sweden)

    Martine Bot

    Full Text Available AIMS: Altered sphingosine 1-phosphate (S1P homeostasis and signaling is implicated in various inflammatory diseases including atherosclerosis. As S1P levels are tightly controlled by S1P lyase, we investigated the impact of hematopoietic S1P lyase (Sgpl1(-/- deficiency on leukocyte subsets relevant to atherosclerosis. METHODS AND RESULTS: LDL receptor deficient mice that were transplanted with Sgpl1(-/- bone marrow showed disrupted S1P gradients translating into lymphopenia and abrogated lymphocyte mitogenic and cytokine response as compared to controls. Remarkably however, Sgpl1(-/- chimeras displayed mild monocytosis, due to impeded stromal retention and myelopoiesis, and plasma cytokine and macrophage expression patterns, that were largely compatible with classical macrophage activation. Collectively these two phenotypic features of Sgpl1 deficiency culminated in diminished atherogenic response. CONCLUSIONS: Here we not only firmly establish the critical role of hematopoietic S1P lyase in controlling S1P levels and T cell trafficking in blood and lymphoid tissue, but also identify leukocyte Sgpl1 as critical factor in monocyte macrophage differentiation and function. Its, partly counterbalancing, pro- and anti-inflammatory activity spectrum imply that intervention in S1P lyase function in inflammatory disorders such as atherosclerosis should be considered with caution.

  11. Sugar-cane juice induces pectin lyase and polygalacturonase in Penicillium griseoroseum

    Directory of Open Access Journals (Sweden)

    Minussi Rosana Cristina

    1998-01-01

    Full Text Available The use of other inducers as substitutes for pectin was studied aiming to reduce the production costs of pectic enzymes. The effects of sugar-cane juice on the production of pectin lyase (PL and polygalacturonase (PG by Penicillium griseoroseum were investigated. The fungus was cultured in a mineral medium (pH 6.3 in a rotary shaker (150 rpm for 48 h at 25oC. Culture media were supplemented with yeast extract and sucrose or sugar-cane juice. Sugar-cane juice added singly to the medium promoted higher PL activity and mycelial dry weight when compared to pectin and the use of sugar-cane juice and yeast extract yielded levels of PG activity that were similar to those obtained with sucrose-yeast extract or pectin. The results indicated that, even at low concentrations, sugar-cane juice was capable of inducing pectin lyase and polygalacturonase with no cellulase activity in P. griseoroseum.

  12. Leader Self Disclosure within PAL: A Case Study

    Science.gov (United States)

    Allen, Adelaide; Court, Sue

    2009-01-01

    The purpose of this paper is to explore the value of self disclosure within Peer Assisted Learning (PAL) sessions at Bournemouth University. We consider the role of self disclosure in education contexts in order to inform our understanding of this skill in PAL. Both qualitative and quantitative methods were employed in this research to investigate…

  13. A Benzothiadiazole Primes Parsley Cells for Augmented Elicitation of Defense Responses

    Science.gov (United States)

    Katz, Vera A.; Thulke, Oliver U.; Conrath, Uwe

    1998-01-01

    Systemic acquired resistance is an important component of the disease-resistance arsenal of plants, and is associated with an enhanced potency for activating local defense responses upon pathogen attack. Here we demonstrate that pretreatment with benzothiadiazole (BTH), a synthetic activator of acquired resistance in plants, augmented the sensitivity for low-dose elicitation of coumarin phytoalexin secretion by cultured parsley (Petroselinum crispum L.) cells. Enhanced coumarin secretion was associated with potentiated activation of genes encoding Phe ammonia-lyase (PAL). The augmentation of PAL gene induction was proportional to the length of pretreatment with BTH, indicating time-dependent priming of the cells. In contrast to the PAL genes, those for anionic peroxidase were directly induced by BTH in the absence of elicitor, thus confirming a dual role for BTH in the activation of plant defenses. Strikingly, the ability of various chemicals to enhance plant disease resistance correlated with their capability to potentiate parsley PAL gene elicitation, emphasizing an important role for defense response potentiation in acquired plant disease resistance. PMID:9701589

  14. Cloning and study of the pectate lyase gene of Erwinia carotovora

    International Nuclear Information System (INIS)

    Bukanov, N.O.; Fonshtein, M.Yu.; Evtushenkov, A.N.; Syarinskii, M.A.; Strel'chenko, P.P.; Yankovski, N.K.; Alikhanyan, S.I.; Fomichev, Yu.K.; Debabov, V.G.

    1986-01-01

    The cloning of the gene of a secretable protein of Erwinia carotovora, pectate lyase, in Escherichia coli was described. Primary cloning was conducted using the phage vector λ 47.1. In the gene library of E. carotovora obtained, eight phages carrying the gene sought were identified according to the appearance of enzymatic activity of the gene product, pectate lyase, in situ. The BamHI fragment of DNA, common to all these phages, was recloned on the plasmid pUC19. It was shown that the cloned pectate lyase gene is represented on the E. carotovora chromosome in one copy. Methods of production of representative gene libraries on phage vectors from no less than 1 μg of cloned DNA even for the genomes of eukaryotes have now been developed. Vectors have been created, for example, λ 47.1, permitting the selection only of hybrid molecules. A number of methods have been developed for the search for a required gene in the library, depending on whether the cloned gene can be expressed or not, and if it can, what properties it will impart to the hybrid clone containing it

  15. Designing technology for emergent literacy: the PictoPal initiative

    NARCIS (Netherlands)

    McKenney, Susan; Voogt, Joke

    2009-01-01

    PictoPal is the name of a technology-supported intervention designed to foster the development of emergent reading and writing skills in four and five year old children. Following the theoretical underpinnings and a brief description of PictoPal, this article describes how children worked with the

  16. Positron annihilation lifetime spectroscopy (PALS): a probe for molecular organisation in self-assembled biomimetic systems.

    Science.gov (United States)

    Fong, Celesta; Dong, Aurelia W; Hill, Anita J; Boyd, Ben J; Drummond, Calum J

    2015-07-21

    Positron annihilation lifetime spectroscopy (PALS) has been shown to be highly sensitive to conformational, structural and microenvironmental transformations arising from subtle geometric changes in molecular geometry in self-assembling biomimetic systems. The ortho-positronium (oPs) may be considered an active probe that can provide information on intrinsic packing and mobility within low molecular weight solids, viscous liquids, and soft matter systems. In this perspective we provide a critical overview of the literature in this field, including the evolution of analysis software and experimental protocols with commentary upon the practical utility of PALS. In particular, we discuss how PALS can provide unique insight into the macroscopic transport properties of several porous biomembrane-like nanostructures and suggest how this insight may provide information on the release of drugs from these matrices to aid in developing therapeutic interventions. We discuss the potentially exciting and fruitful application of this technique to membrane dynamics, diffusion and permeability. We propose that PALS can provide novel molecular level information that is complementary to conventional characterisation techniques.

  17. Positron annihilation lifetime spectroscopy (PALS) as a characterization technique for nanostructured self-assembled amphiphile systems.

    Science.gov (United States)

    Dong, Aurelia W; Pascual-Izarra, Carlos; Pas, Steven J; Hill, Anita J; Boyd, Ben J; Drummond, Calum J

    2009-01-08

    Positron annihilation lifetime spectroscopy (PALS) has potential as a novel rapid characterization method for self-assembly amphiphile systems; however, a lack of systematic correlation of PALS parameters with structural attributes has limited its more widespread application. In this study, using the well-characterized phytantriol/water and the phytantriol/vitamin E acetate/water self-assembly amphiphile systems, the impact of systematic structural changes controlled by changes in composition and temperature on PALS parameters has been studied. The PALS parameters (orthopositronium (oPs) lifetime and intensity signatures) were shown to be sensitive to the molecular packing and mobility of the self-assembled lipid molecules in various lyotropic liquid crystalline phases, enabling differentiation between liquid crystalline structures. The oPs lifetime, related to the molecular packing and mobility, is correlated with rheological properties of the individual mesophases. The oPs lifetime links the lipid chain packing and mobility in the various mesophases to resultant macroscopic properties, such as permeability, which is critical for the use of these mesophase structures as diffusion-controlled release matrices for active liposoluble compounds.

  18. Abscisic Acid Induced Changes in Production of Primary and Secondary Metabolites, Photosynthetic Capacity, Antioxidant Capability, Antioxidant Enzymes and Lipoxygenase Inhibitory Activity of Orthosiphon stamineus Benth.

    Directory of Open Access Journals (Sweden)

    Mohd Hafiz Ibrahim

    2013-07-01

    Full Text Available An experiment was conducted to investigate and distinguish the relationships in the production of total phenolics, total flavonoids, soluble sugars, H2O2, O2−, phenylalanine ammonia lyase (PAL activity, leaf gas exchange, antioxidant activity, antioxidant enzyme activity [ascorbate peroxidase (APX, catalase (CAT, superoxide dismutase (SOD and Lipoxygenase inhibitory activity (LOX] under four levels of foliar abscisic acid (ABA application (0, 2, 4, 6 µM for 15 weeks in Orthosiphon stamineus Benth. It was found that the production of plant secondary metabolites, soluble sugars, antioxidant activity, PAL activity and LOX inhibitory activity was influenced by foliar application of ABA. As the concentration of ABA was increased from 0 to 6 µM the production of total phenolics, flavonoids, sucrose, H2O2, O2−, PAL activity and LOX inhibitory activity was enhanced. It was also observed that the antioxidant capabilities (DPPH and ORAC were increased. This was followed by increases in production of antioxidant enzymes APX, CAT and SOD. Under high application rates of ABA the net photosynthesis and stomatal conductance was found to be reduced. The production of primary and secondary metabolites displayed a significant positive relationship with H2O2 (total phenolics, r2 = 0.877; total flavonoids, r2 = 0.812; p ≤ 0.05 and O2− (total phenolics, r2 = 0.778; total flavonoids, r2 = 0.912; p ≤ 0.05. This indicated that increased oxidative stress at high application rates of ABA, improved the production of phytochemicals.

  19. BitPAl: a bit-parallel, general integer-scoring sequence alignment algorithm.

    Science.gov (United States)

    Loving, Joshua; Hernandez, Yozen; Benson, Gary

    2014-11-15

    Mapping of high-throughput sequencing data and other bulk sequence comparison applications have motivated a search for high-efficiency sequence alignment algorithms. The bit-parallel approach represents individual cells in an alignment scoring matrix as bits in computer words and emulates the calculation of scores by a series of logic operations composed of AND, OR, XOR, complement, shift and addition. Bit-parallelism has been successfully applied to the longest common subsequence (LCS) and edit-distance problems, producing fast algorithms in practice. We have developed BitPAl, a bit-parallel algorithm for general, integer-scoring global alignment. Integer-scoring schemes assign integer weights for match, mismatch and insertion/deletion. The BitPAl method uses structural properties in the relationship between adjacent scores in the scoring matrix to construct classes of efficient algorithms, each designed for a particular set of weights. In timed tests, we show that BitPAl runs 7-25 times faster than a standard iterative algorithm. Source code is freely available for download at http://lobstah.bu.edu/BitPAl/BitPAl.html. BitPAl is implemented in C and runs on all major operating systems. jloving@bu.edu or yhernand@bu.edu or gbenson@bu.edu Supplementary data are available at Bioinformatics online. © The Author 2014. Published by Oxford University Press.

  20. Early phenylpropanoid biosynthetic steps in Cannabis sativa: link between genes and metabolites.

    Science.gov (United States)

    Docimo, Teresa; Consonni, Roberto; Coraggio, Immacolata; Mattana, Monica

    2013-06-28

    Phenylalanine ammonia-lyase (PAL), Cinnamic acid 4-hydroxylase (C4H) and 4-Coumarate: CoA ligase (4CL) catalyze the first three steps of the general phenylpropanoid pathway whereas chalcone synthase (CHS) catalyzes the first specific step towards flavonoids production. This class of specialized metabolites has a wide range of biological functions in plant development and defence and a broad spectrum of therapeutic activities for human health. In this study, we report the isolation of hemp PAL and 4CL cDNA and genomic clones. Through in silico analysis of their deduced amino acid sequences, more than an 80% identity with homologues genes of other plants was shown and phylogenetic relationships were highlighted. Quantitative expression analysis of the four above mentioned genes, PAL and 4CL enzymatic activities, lignin content and NMR metabolite fingerprinting in different Cannabis sativa tissues were evaluated. Furthermore, the use of different substrates to assay PAL and 4CL enzymatic activities indicated that different isoforms were active in different tissues. The diversity in secondary metabolites content observed in leaves (mainly flavonoids) and roots (mainly lignin) was discussed in relation to gene expression and enzymatic activities data.

  1. Early Phenylpropanoid Biosynthetic Steps in Cannabis sativa: Link between Genes and Metabolites

    Directory of Open Access Journals (Sweden)

    Immacolata Coraggio

    2013-06-01

    Full Text Available Phenylalanine ammonia-lyase (PAL, Cinnamic acid 4-hydroxylase (C4H and 4-Coumarate: CoA ligase (4CL catalyze the first three steps of the general phenylpropanoid pathway whereas chalcone synthase (CHS catalyzes the first specific step towards flavonoids production. This class of specialized metabolites has a wide range of biological functions in plant development and defence and a broad spectrum of therapeutic activities for human health. In this study, we report the isolation of hemp PAL and 4CL cDNA and genomic clones. Through in silico analysis of their deduced amino acid sequences, more than an 80% identity with homologues genes of other plants was shown and phylogenetic relationships were highlighted. Quantitative expression analysis of the four above mentioned genes, PAL and 4CL enzymatic activities, lignin content and NMR metabolite fingerprinting in different Cannabis sativa tissues were evaluated. Furthermore, the use of different substrates to assay PAL and 4CL enzymatic activities indicated that different isoforms were active in different tissues. The diversity in secondary metabolites content observed in leaves (mainly flavonoids and roots (mainly lignin was discussed in relation to gene expression and enzymatic activities data.

  2. Characterization of two bacterial hydroxynitrile lyases with high similarity to cupin superfamily proteins

    NARCIS (Netherlands)

    Hussain, Z.; Wiedner, R.; Steiner, K.; Hajek, T.; Avi, M.; Hecher, B.; Sessitsch, A.; Schwab, H.

    2012-01-01

    Hydroxynitrile lyases (HNLs) catalyze the cleavage of cyanohydrins. In the reverse reaction, they catalyze the formation of carbon-carbon bonds by enantioselective condensation of hydrocyanic acid with carbonyls. In this study, we describe two proteins from endophytic bacteria that display activity

  3. The replicative DNA polymerase of herpes simplex virus 1 exhibits apurinic/apyrimidinic and 5′-deoxyribose phosphate lyase activities

    OpenAIRE

    Bogani, Federica; Boehmer, Paul E.

    2008-01-01

    Base excision repair (BER) is essential for maintaining genome stability both to counter the accumulation of unusual bases and to protect from base loss in the DNA. Herpes simplex virus 1 (HSV-1) is a large dsDNA virus that encodes its own DNA replication machinery, including enzymes involved in nucleotide metabolism. We report on a replicative family B and a herpesvirus-encoded DNA Pol that possesses DNA lyase activity. We have discovered that the catalytic subunit of the HSV-1 DNA polymeras...

  4. Molecular Cloning of cpcU and Heterodimeric Bilin Lyase Activity Analysis of CpcU and CpcS for Attachment of Phycocyanobilin to Cys-82 on the β-Subunit of Phycocyanin in Arthrospira platensis FACHB314

    Directory of Open Access Journals (Sweden)

    Fei Wu

    2016-03-01

    Full Text Available A new bilin lyase gene cpcU was cloned from Arthrospira platensis FACHB314 to study the assembly of the phycocyanin β-Subunit. Two recombinant plasmids, one contained the phycocyanobilin (PCB producing genes (hoxI and pcyA, while the other contained the gene of the β-Subunit of phycobiliprotein (cpcB and the lyase gene (cpcU, cpcS, or cpcU/S were constructed and separately transferred into Escherichia coli in order to test the activities of relevant lyases for catalyzing PCB addition to CpcB during synthesizing fluorescent β-PC of A. platensis FACHB314. The fluorescence intensity examination showed that Cys-82 maybe the active site for the β-Subunit binding to PCBs and the attachment could be carried out by CpcU, CpcS, or co-expressed cpcU/S in A. platensis FACHB314.

  5. A 5-methylcytosine DNA glycosylase/lyase demethylates the retrotransposon Tos17 and promotes its transposition in rice

    KAUST Repository

    La, Honggui; Ding, Bo; Mishra, Gyan Prakash; Zhou, Bo; Yang, Hongmei; Bellizzi, Maria Del Rosario; Chen, Songbiao; Meyers, Blake C.; Peng, Zhaohua; Zhu, Jian-Kang; Wang, Guoliang

    2011-01-01

    DNA 5-methylcytosine (5-meC) is an important epigenetic mark for transcriptional gene silencing in many eukaryotes. In Arabidopsis, 5-meC DNA glycosylase/lyases actively remove 5-meC to counter-act transcriptional gene silencing in a locus-specific manner, and have been suggested to maintain the expression of transposons. However, it is unclear whether plant DNA demethylases can promote the transposition of transposons. Here we report the functional characterization of the DNA glycosylase/lyase DNG701 in rice. DNG701 encodes a large (1,812 amino acid residues) DNA glycosylase domain protein. Recombinant DNG701 protein showed 5-meC DNA glycosylase and lyase activities in vitro. Knockout or knockdown of DNG701 in rice plants led to DNA hypermethylation and reduced expression of the retrotransposon Tos17. Tos17 showed less transposition in calli derived from dng701 knockout mutant seeds compared with that in wild-type calli. Overexpression of DNG701 in both rice calli and transgenic plants substantially reduced DNA methylation levels of Tos17 and enhanced its expression. The overexpression also led to more frequent transposition of Tos17 in calli. Our results demonstrate that rice DNG701 is a 5-meC DNA glycosylase/lyase responsible for the demethylation of Tos17 and this DNA demethylase plays a critical role in promoting Tos17 transposition in rice calli.

  6. A 5-methylcytosine DNA glycosylase/lyase demethylates the retrotransposon Tos17 and promotes its transposition in rice

    KAUST Repository

    La, Honggui

    2011-09-06

    DNA 5-methylcytosine (5-meC) is an important epigenetic mark for transcriptional gene silencing in many eukaryotes. In Arabidopsis, 5-meC DNA glycosylase/lyases actively remove 5-meC to counter-act transcriptional gene silencing in a locus-specific manner, and have been suggested to maintain the expression of transposons. However, it is unclear whether plant DNA demethylases can promote the transposition of transposons. Here we report the functional characterization of the DNA glycosylase/lyase DNG701 in rice. DNG701 encodes a large (1,812 amino acid residues) DNA glycosylase domain protein. Recombinant DNG701 protein showed 5-meC DNA glycosylase and lyase activities in vitro. Knockout or knockdown of DNG701 in rice plants led to DNA hypermethylation and reduced expression of the retrotransposon Tos17. Tos17 showed less transposition in calli derived from dng701 knockout mutant seeds compared with that in wild-type calli. Overexpression of DNG701 in both rice calli and transgenic plants substantially reduced DNA methylation levels of Tos17 and enhanced its expression. The overexpression also led to more frequent transposition of Tos17 in calli. Our results demonstrate that rice DNG701 is a 5-meC DNA glycosylase/lyase responsible for the demethylation of Tos17 and this DNA demethylase plays a critical role in promoting Tos17 transposition in rice calli.

  7. Molecular cloning, characterization and expression of phenylalanine ...

    African Journals Online (AJOL)

    A full-length cDNA and genomic DNA of phenylalanine ammonia-lyase gene, which catalyzes the first step in the flavonoid biosynthetic pathway, were isolated from Ginkgo biloba for the first time (designated as GbPAL, GenBank Accession No. EU071050). The cDNA and genomic DNA sequences of GbPAL were the same, ...

  8. Structure and Mechanism of PhnP, a Phosphodiesterase of the Carbon-Phosphorus Lyase Pathway

    DEFF Research Database (Denmark)

    He, Shu-Mei; Wathier, Matthew; Podzelinska, Kateryna

    2011-01-01

    PhnP is a phosphodiesterase that plays an important role within the bacterial carbon-phosphorus lyase (CP-lyase) pathway by recycling a "dead-end" intermediate, 5-phospho-α-d-ribosyl 1,2-cyclic phosphate, that is formed during organophosphonate catabolism. As a member of the metallo-β-lactamase s......PhnP is a phosphodiesterase that plays an important role within the bacterial carbon-phosphorus lyase (CP-lyase) pathway by recycling a "dead-end" intermediate, 5-phospho-α-d-ribosyl 1,2-cyclic phosphate, that is formed during organophosphonate catabolism. As a member of the metallo...

  9. High-Level Expression of a Thermally Stable Alginate Lyase Using Pichia pastoris, Characterization and Application in Producing Brown Alginate Oligosaccharide

    Directory of Open Access Journals (Sweden)

    Haifeng Li

    2018-05-01

    Full Text Available An alginate lyase encoding gene sagl from Flavobacterium sp. H63 was codon optimized and recombinantly expressed at high level in P.pastoris through high cell-density fermentation. The highest yield of recombinant enzyme of sagl (rSAGL in yeast culture supernatant reached 226.4 μg/mL (915.5 U/mL. This was the highest yield record of recombinant expression of alginate lyase so far. The rSAGL was confirmed as a partially glycosylated protein through EndoH digestion. The optimal reaction temperature and pH of this enzyme were 45 °C and 7.5; 80 mM K+ ions could improve the catalytic activity of the enzyme by 244% at most. rSAGL was a thermal stable enzyme with T5015 of 57–58 °C and T5030 of 53–54 °C. Its thermal stability was better than any known alginate lyase. In 100 mM phosphate buffer of pH 6.0, rSAGL could retain 98.8% of the initial activity after incubation at 50 °C for 2 h. Furthermore, it could retain 61.6% of the initial activity after 48 h. The specific activity of the purified rSAGL produced by P. pastoris attained 4044 U/mg protein, which was the second highest record of alginate lyase so far. When the crude enzyme of the rSAGL was directly used in transformation of sodium alginate with 40 g/L, 97.2% of the substrate was transformed to di, tri, tetra brown alginate oligosaccharide after 32 h of incubation at 50 °C, and the final concentration of reducing sugar in mixture reached 9.51 g/L. This is the first report of high-level expression of thermally stable alginate lyase using P. pastoris system.

  10. Prenatal diagnosis in adenylosuccinate lyase deficiency

    NARCIS (Netherlands)

    Marie, S.; Flipsen, J. W.; Duran, M.; Poll-The, B. T.; Beemer, F. A.; Bosschaart, A. N.; Vincent, M. F.; van den Berghe, G.

    2000-01-01

    Adenylosuccinate lyase deficiency, an autosomal recessive inborn error of purine synthesis, provokes accumulation in body fluids of succinylaminoimidazolecarboxamide riboside and succinyladenosine, the dephosphorylated derivatives of the two substrates of the enzyme. Most patients display severe

  11. Neuroprotective effects of (Val8)GLP-1-Glu-PAL in the MPTP Parkinson's disease mouse model.

    Science.gov (United States)

    Zhang, YanFang; Chen, YiMei; Li, Lin; Hölscher, Christian

    2015-10-15

    Glucagon-like peptide 1 (GLP-1) is a hormone and a growth factor. GLP-1 mimetics are currently on the market as treatments for type 2 diabetes. They also have shown neuroprotective properties in animal models of neurodegenerative disorders. In addition, the GLP-1 mimetic exendin-4 has shown protective effects in animal models of Parkinson's disease (PD), and a first clinical trial in PD patients showed promising results. (Val8)GLP-1-glu-PAL is a new GLP-1 analogue which has a longer biological half-life than exendin-4. We previously showed that (Val8)GLP-1-glu-PAL has neuroprotective properties. Here we tested the drug in the 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) mouse model of PD. MPTP was injected (30mg/kg i.p.) along with (Val8)GLP-1-glu-PAL (25nmol/kg i.p.) once-daily for 8 days. (Val8)GLP-1-glu-PAL showed good effects in preventing the MPTP-induced motor impairment (Rotarod, open field locomotion, swim test), reduction in tyrosine hydroxylase levels (dopamine synthesis) in the substantia nigra, a reduction of activated caspase 3 levels, of TUNEL positive cell numbers, of the pro-apoptotic signaling molecule BAX and an increase in the growth signaling molecule Bcl-2. The results demonstrate that (Val8)GLP-1-glu-PAL shows promise as a novel treatment of PD. Copyright © 2015 Elsevier B.V. All rights reserved.

  12. How much locomotive activity is needed for an active physical activity level: analysis of total step counts

    Directory of Open Access Journals (Sweden)

    Ohkawara Kazunori

    2011-11-01

    Full Text Available Abstract Background Although physical activity recommendations for public health have focused on locomotive activity such as walking and running, it is uncertain how much these activities contribute to overall physical activity level (PAL. The purpose of the present study was to determine the contribution of locomotive activity to PAL using total step counts measured in a calorimeter study. Methods PAL, calculated as total energy expenditure divided by basal metabolic rate, was evaluated in 11 adult men using three different conditions for 24-hour human calorimeter measurements: a low-activity day (L-day targeted at a low active level of PAL (1.45, and a high-frequency moderate activity day (M-day or a high-frequency vigorous activity day (V-day targeted at an active level of PAL (1.75. These subjects were permitted only light activities except prescribed activities. In a separate group of 41 adults, free-living PAL was evaluated using doubly-labeled water (DLW. In both experiments, step counts per day were also measured using an accelerometer. Results In the human calorimeter study, PAL and step counts were 1.42 ± 0.10 and 8,973 ± 543 steps/d (L-day, 1.82 ± 0.14 and 29,588 ± 1,126 steps/d (M-day, and 1.74 ± 0.15 and 23,755 ± 1,038 steps/d (V-day, respectively. In the DLW study, PAL and step counts were 1.73 ± 0.15 and 10,022 ± 2,605 steps/d, and there was no significant relationship between PAL and daily step counts. Conclusions These results indicate that an enormous number of steps are needed for an active level of PAL if individuals extend physical activity-induced energy expenditure by only locomotive activity. Therefore, non-locomotive activity such as household activity should also play a significant role in increasing PAL under free-living conditions.

  13. A novel gene encoding xanthan lyase of Paenibacillus alginolyticus strain XL-1

    NARCIS (Netherlands)

    Ruijssenaars, H.J.; Hartmans, S.; Verdoes, J.C.

    2000-01-01

    Xanthan-modifying enzymes are powerful tools in studying structure-function relationships of this polysaccharide. One of these modifying enzymes is xanthan lyase, which removes the terminal side chain residue of xanthan. In this paper, the cloning and sequencing of the first xanthan lyase-encoding

  14. RESPONSE OF PHENOLIC METABOLISM INDUCED BY ALUMINIUM TOXICITY IN FAGOPYRUM ESCULENTUM MOENCH. PLANTS.

    Science.gov (United States)

    Smirnov, O E; Kosyan, A M; Kosyk, O I; Taran, N Yu

    2015-01-01

    Buckwheat genus (Fagopyrum Mill.) is one of the aluminium tolerant taxonomic units of plants. The aim of the study was an evaluation of the aluminium (50 μM effect on phenolic accumulation in various parts of buckwheat plants (Fagopyrum esculentum Moench). Detection of increasing of total phenolic content, changes in flavonoid and anthocyanin content and phenylalanine ammonia-lyase activity (PAL) were revealed over a period of 10 days of exposure to aluminium. The most significant effects of aluminium treatment on phenolic compounds accumulation were total phenolic content increasing (by 27.2%) and PAL activity rising by 2.5 times observed in leaves tissues. Received data could be helpful to understand the aluminium tolerance principles and relationships of phenolic compounds to aluminium phytotoxicity.

  15. Crystallization and preliminary X-ray analysis of βC–S lyases from two oral streptococci

    International Nuclear Information System (INIS)

    Kezuka, Yuichiro; Yoshida, Yasuo; Nonaka, Takamasa

    2009-01-01

    The βC-S lyases from two oral bacteria, Streptococcus anginosus and S. gordonii, were cloned, overproduced, purified and crystallized. The obtained crystals were characterized by X-ray diffraction. Hydrogen sulfide, which causes oral malodour, is generally produced from l-cysteine by the action of βC–S lyase from oral bacteria. The βC–S lyases from two oral bacteria, Streptococcus anginosus and S. gordonii, have been cloned, overproduced, purified and crystallized. X-ray diffraction data were collected from the two types of crystals using synchrotron radiation. The crystal of S. anginosus βC–S lyase belonged to the orthorhombic space group P2 1 2 1 2 1 , with unit-cell parameters a = 67.0, b = 111.1, c = 216.4 Å, and the crystal of S. gordonii βC–S lyase belonged to the same space group, with unit-cell parameters a = 58.0, b = 73.9. c = 187.6 Å. The structures of the βC–S lyases were solved by molecular-replacement techniques

  16. Mechanism of Cytochrome P450 17A1-Catalyzed Hydroxylase and Lyase Reactions

    DEFF Research Database (Denmark)

    Bonomo, Silvia; Jorgensen, Flemming Steen; Olsen, Lars

    2017-01-01

    Cytochrome P450 17A1 (CYP17A1) catalyzes C17 hydroxylation of pregnenolone and progesterone and the subsequent C17–C20 bond cleavage (lyase reaction) to form androgen precursors. Compound I (Cpd I) and peroxo anion (POA) are the heme-reactive species underlying the two reactions. We have characte...... the concept that the selectivity of the steroidogenic CYPs is ruled by direct interactions with the enzyme, in contrast to the selectivity of drug-metabolizing CYPs, where the reactivity of the substrates dominates....... characterized the reaction path for both the hydroxylase and lyase reactions using density functional theory (DFT) calculations and the enzyme–substrate interactions by molecular dynamics (MD) simulations. Activation barriers for positions subject to hydroxylase reaction have values close to each other and span...

  17. Cytochrome c and c1 heme lyases are essential in Plasmodium berghei.

    Science.gov (United States)

    Posayapisit, Navaporn; Songsungthong, Warangkhana; Koonyosying, Pongpisid; Falade, Mofolusho O; Uthaipibull, Chairat; Yuthavong, Yongyuth; Shaw, Philip J; Kamchonwongpaisan, Sumalee

    Malaria parasites possess a de novo heme synthetic pathway. Interestingly, this pathway is dispensable during the blood stages of development in mammalian hosts. The assembly of the two most important hemeproteins, cytochromes c and c1, is mediated by cytochrome heme lyase enzymes. Plasmodium spp. possess two cytochrome heme lyases encoded by separate genes. Given the redundancy of heme synthesis, we sought to determine if heme lyase function also exhibits redundancy. To answer this question, we performed gene knockout experiments. We found that the PBANKA_143950 and PBANKA_0602600 Plasmodium berghei genes encoding cytochrome c (Pbcchl) and cytochrome c1 (Pbcc 1 hl) heme lyases, respectively, can only be disrupted when a complementary gene is present. In contrast, four genes in the de novo heme synthesis pathway can be disrupted without complementation. This work provides evidence that Pbcchl and Pbcc 1 hl are both essential and thus may be antimalarial targets. Copyright © 2016 Elsevier B.V. All rights reserved.

  18. Mixed-methods development of a new patient-reported outcome instrument for chronic low back pain: part 1-the Patient Assessment for Low Back Pain - Symptoms (PAL-S).

    Science.gov (United States)

    Martin, Mona L; Blum, Steven I; Liedgens, Hiltrud; Bushnell, Donald M; McCarrier, Kelly P; Hatley, Noël V; Ramasamy, Abhilasha; Freynhagen, Rainer; Wallace, Mark; Argoff, Charles; Eerdekens, Mariёlle; Kok, Maurits; Patrick, Donald L

    2018-06-01

    We describe the mixed-methods (qualitative and quantitative) development and preliminary validation of the Patient Assessment for Low Back Pain-Symptoms (PAL-S), a patient-reported outcome measure for use in chronic low back pain (cLBP) clinical trials. Qualitative methods (concept elicitation and cognitive interviews) were used to identify and refine symptom concepts and quantitative methods (classical test theory and Rasch measurement theory) were used to evaluate item- and scale-level performance of the measure using an iterative approach. Patients with cLBP participated in concept elicitation interviews (N = 43), cognitive interviews (N = 38), and interview-based assessment of paper-to-electronic mode equivalence (N = 8). A web-based sample of patients with self-reported cLBP participated in quantitative studies to evaluate preliminary (N = 598) and revised (n = 401) drafts and a physician-diagnosed cohort of patients with cLBP (N = 45) participated in preliminary validation of the measure. The PAL-S contained 14 items describing symptoms (overall pain, sharp, prickling, sensitive, tender, radiating, shocking, shooting, burning, squeezing, muscle spasms, throbbing, aching, and stiffness). Item-level performance, scale structure, and scoring seemed to be appropriate. One-week test-retest reproducibility was acceptable (intraclass correlation coefficient 0.81 [95% confidence interval, 0.61-0.91]). Convergent validity was demonstrated with total score and MOS-36 Bodily Pain (Pearson correlation -0.79), Neuropathic Pain Symptom Inventory (0.73), Roland-Morris Disability Questionnaire (0.67), and MOS-36 Physical Functioning (-0.65). Individual item scores and total score discriminated between numeric rating scale tertile groups and painDETECT categories. Respondent interpretation of paper and electronic administration modes was equivalent. The PAL-S has demonstrated content validity and is potentially useful to assess treatment benefit in cLBP clinical trials.

  19. Retrieving soil moisture for non-forested areas using PALS radiometer measurements in SMAPVEX12 field campaign

    Science.gov (United States)

    In this paper we investigate retrieval of soil moisture based on L-band brightness temperature under diverse conditions and land cover types. We apply the PALS (Passive Active L-band System) radiometer data collected in the SMAPVEX12 (Soil Moisture Active Passive Validation Experiment 2012) field ex...

  20. Dose and time-dependent effects of cyanide on thiosulfate sulfurtransferase, 3-mercaptopyruvate sulfurtransferase, and cystathionine λ-lyase activities.

    Science.gov (United States)

    Singh, Poonam; Rao, Pooja; Bhattacharya, Rahul

    2013-12-01

    We assessed the dose-dependent effect of potassium cyanide (KCN) on thiosulfate sulfurtransferase (TST), 3-mercaptopyruvate sulfurtransferase (3-MPST), and cystathionine λ-lyase (CST) activities in mice. The time-dependent effect of 0.5 LD50 KCN on cyanide level and cytochrome c oxidase (CCO), TST, 3-MPST, and CST activities was also examined. Furthermore, TST, 3-MPST, and CST activities were measured in stored mice cadavers. Hepatic and renal TST activity increased by 0.5 LD50 KCN but diminished by ≥2.0 LD50. After 0.5 LD50 KCN, the elevated hepatic cyanide level was accompanied by increased TST, 3-MPST, and CST activities, and CCO inhibition. Elevated renal cyanide level was only accompanied by increased 3-MPST activity. No appreciable change in enzyme activities was observed in mice cadavers. The study concludes that high doses of cyanide exert saturating effects on its detoxification enzymes, indicating their exogenous use during cyanide poisoning. Also, these enzymes are not reliable markers of cyanide poisoning in autopsied samples. © 2013 Wiley Periodicals, Inc.

  1. Novel Pectate Lyase Genes of Heterodera glycines Play Key Roles in the Early Stage of Parasitism.

    Directory of Open Access Journals (Sweden)

    Huan Peng

    Full Text Available Pectate lyases are known to play a key role in pectin degradation by catalyzing the random cleavage of internal polymer linkages (endo-pectinases. In this paper, four novel cDNAs, designated Hg-pel-3, Hg-pel-4, Hg-pel-6 and Hg-pel-7, that encode pectate lyases were cloned and characterized from the soybean cyst nematode, Heterodera glycines. The predicted protein sequences of HG-PEL-3, HG-PEL-4 and HG-PEL-6 differed significantly in both their amino acid sequences and their genomic structures from other pectate lyases of H. glycines (HG-PEL-1, HG-PEL-2 and HG-PEL-7. A phylogenetic study revealed that the pectate lyase proteins of H. glycines are clustered into distinct clades and have distinct numbers and positioning of introns, which suggests that the pectate lyase genes of H. glycines may have evolved from at least two ancestral genes. A Southern blot analysis revealed that multiple Hg-pel-6-like genes were present in the H. glycines genome. In situ hybridization showed that four novel pectate lyases (Hg-pel-3, Hg-pel-4, Hg-pel-6 and Hg-pel-7 were actively transcribed in the subventral esophageal gland cells. A semi-quantitative RT-PCR assay supported the finding that the expression of these genes was strong in the egg, pre-parasitic second-stage juvenile (J2 and early parasitic J2 stages and that it declined in further developmental stages of the nematode. This expression pattern suggests that these proteins play a role in the migratory phase of the nematode life cycle. Knocking down Hg-pel-6 using in vitro RNA interference resulted in a 46.9% reduction of the number of nematodes that invaded the plants and a 61.5% suppression of the development of H. glycines females within roots compared to the GFP-dsRNA control. Plant host-derived RNAi induced the silencing of the Hg-pel-6gene, which significantly reduced the nematode infection levels at 7 Days post inoculation (dpi. Similarly, this procedure reduced the number of female adults at 40 dpi

  2. Purification and Characterization of a Unique Pectin Lyase from Aspergillus giganteus Able to Release Unsaturated Monogalacturonate during Pectin Degradation

    Directory of Open Access Journals (Sweden)

    Danielle Biscaro Pedrolli

    2014-01-01

    Full Text Available A pectin lyase, named PLIII, was purified to homogeneity from the culture filtrate of Aspergillus giganteus grown in submerged culture containing orange peel waste as carbon source. PLIII was able to digest apple pectin and citrus pectins with different degrees of methyl esterification. Interestingly, the PLIII activity was stimulated in the presence of some divalent cations including Pb2+ and was not significantly affected by Hg2+. Like other pectin lyases, PLIII is stimulated by but is not dependent on Ca2+. The main soluble product released during the degradation of pectic substances promoted by the PLIII is compatible with an unsaturated monogalacturonate. PLIII is a unique enzyme able to release unsaturated monogalacturonate as the only soluble product during the degradation of pectic substances; therefore, PLIII was classified as an exo-pectin lyase. To our knowledge, this is the first characterization of an exo-pectin lyase. The PLIII described in this work is potentially useful for ethanol production from pectin-rich biomass, besides other common applications for alkaline pectinases like preparation of textile fibers, coffee and tea fermentation, vegetable oil extraction, and the treatment of pulp in papermaking.

  3. PENGARUH PENERAPAN STRATEGI PEER ASSISTED LEARNING (PAL TERHADAP KEMAMPUAN LITERASI SAINS MAHASISWA DALAM PERKULIAHAN MORFOLOGI TUMBUHAN

    Directory of Open Access Journals (Sweden)

    Sariwulan Diana

    2016-11-01

    Telah dilakukan penelitian tentang pengaruh penerapan strategi Peer Assisted Learning (PAL dalam perkuliahan Morfologi Tumbuhan, yang bertujuan untuk meningkatkan kemampuan literasi sains mahasiswa. Penelitian ini menggunakan metode eksperimen, dengan rancangan quasi eksperimental jenis pretest postest control group design. Rancangan tersebut mencakup satu kelompok mahasiswa yang diberi pretes (tes yang dilakukan sebelum perlakuan PAL yang kemudian dilanjutkan dengan mengobservasi proses perlakuan PAL dan diakhiri postes. Satu kelompok mahasiswa lainnya (kontrol hanya diberi pretes dan diakhiri postes. Instrumen penelitian meliputi angket untuk menjaring tutor, seperangkat soal pretes, soal responsi, soal postes dan angket tentang respon mahasiswa terhadap program PAL yang dilakukan. Penerapan strategi PAL berpengaruh signifikan terhadap peningkatan kemampuan literasi sains mahasiswa dalam  perkuliahan Morfologi Tumbuhan (thitung > ttabel. Peningkatan literasi sains mahasiswa pada kelas eksperimen termasuk kategori sedang (N-gain 0,4 dan rendah (N-gain 0,1 pada kelas kontrol. Semua mahasiswa merespon positif terhadap startegi PAL yang dilakukan.

  4. Chondroitin Sulfate (CS) Lyases: Structure, Function and Application in Therapeutics.

    Science.gov (United States)

    Rani, Aruna; Patel, Seema; Goyal, Arun

    2018-01-01

    Glycosaminoglycans (GAGs) such as chondroitin sulfate (CS) are the chief natural polysaccharides which reside in biological tissues mainly in extracellular matrix. These CS along with adhesion molecules and growth factors are involved in central nervous system (CNS) development, cell progression and pathogenesis. The chondroitin lyases are the enzyme that degrade and alter the CS chains and hence modify various signalling pathways involving CS chains. These CS lyases are substrate specific, can precisely manipulate the CS polysaccharides and have various biotechnological, medical and therapeutic applications. These enzymes can be used to produce the unsaturated oligosaccharides, which have immune-modulatory, anti-inflammatory and neuroprotective properties. This review focuses on the major breakthrough of the chondroitin sulfate degrading enzymes, their structures and functioning mechanism. This also provides comprehensive information regarding production, purification, characterization of CS lyases and their major applications, both established as well as emerging ones such as neural development. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

  5. StarPals International Young Astronomers' Network Collaborative Projects for IYA

    Science.gov (United States)

    Kingan, Jessi

    2008-09-01

    StarPals is a nascent non-profit organization with the goal of providing opportunities for international collaboration between students of all ages within space science research. We believe that by encouraging an interest in the cosmos, the one thing that is truly Universal, from a young age, students will not only further their knowledge of and interest in science but will learn valuable teamwork and life skills. The goal is to foster respect, understanding and appreciation of cultural diversity among all StarPals participants, whether students, teachers, or mentors. StarPals aims to inspire students by providing opportunities in which, more than simply visualizing themselves as research scientists, they can actually become one. The technologies of robotic telescopes, videoconferencing, and online classrooms are expanding the possibilities like never before. In honor of IYA2009, StarPals would like to encourage 400 schools to participate on a global scale in astronomy/cosmology research on various concurrent projects. We will offer in-person or online workshops and training sessions to teach the teachers. We will be seeking publication in scientific journals for some student research. For our current project, the Double Stars Challenge, students use the robotic telescopes to take a series of four images of one of 30 double stars from a list furnished by the US Naval Observatory and then use MPO Canopus software to take distance and position angle measurements. StarPals provides students with hands-on training, telescope time, and software to complete the imaging and measuring. A paper will be drafted from our research data and submitted to the Journal of Double Star Observations. The kids who participate in this project may potentially be the youngest contributors to an article in a vetted scientific journal. Kids rapidly adapt and improve their computer skills operating these telescopes and discover for themselves that science is COOL!

  6. Preparation of 1D Hierarchical Material Mesosilica/Pal Composite and Its Performance in the Adsorption of Methyl Orange.

    Science.gov (United States)

    Wu, Mei; Han, Haifeng; Ni, Lingli; Song, Daiyun; Li, Shuang; Hu, Tao; Jiang, Jinlong; Chen, Jing

    2018-01-20

    This paper highlights the synthesis of a one-dimensional (1D) hierarchical material mesosilica/palygorskite (Pal) composite and evaluates its adsorption performance for anionic dye methyl orange (MO) in comparison with Pal and Mobile crystalline material-41 (MCM-41). The Mesosilica/Pal composite is consisted of mesosilica coated Pal nanorods and prepared through a dual template approach using cetyltrimethyl ammonium bromide (CTAB) and Pal as soft and hard templates, respectively. The composition and structure of the resultant material was characterized by a scanning electron microscope (SEM), transmissionelectron microscopy (TEM), N₂ adsorption-desorption analysis, small-angle X-Ray powder diffraction (XRD), and zeta potential measurement. Adsorption experiments were carried out with different absorbents at different contact times and pH levels. Compared with Pal and MCM-41, the mesosilica/Pal composite exhibited the best efficiency for MO adsorption. Its adsorption ratio is as high as 70.4%. Its adsorption equilibrium time is as short as 30 min. Results testify that the MO retention is promoted for the micro-mesoporous hierarchical structure and positive surface charge electrostatic interactions of the mesosilica/Pal composite. The regenerability of the mesosilica/Pal composite absorbent was also assessed. 1D morphology makes it facile to separate from aqueous solutions. It can be effortlessly recovered and reused for up to nine cycles.

  7. Construction and Commissioning of PAL-XFEL Facility

    Directory of Open Access Journals (Sweden)

    In Soo Ko

    2017-05-01

    Full Text Available The construction of Pohang Accelerator Laboratory X-ray Free-Electron Laser (PAL-XFEL, a 0.1-nm hard X-ray free-electron laser (FEL facility based on a 10-GeV S-band linear accelerator (LINAC, is achieved in Pohang, Korea by the end of 2016. The construction of the 1.11 km-long building was completed by the end of 2014, and the installation of the 10-GeV LINAC and undulators started in January 2015. The installation of the 10-GeV LINAC, together with the undulators and beamlines, was completed by the end of 2015. The commissioning began in April 2016, and the first lasing of the hard X-ray FEL line was achieved on 14 June 2016. The progress of the PAL-XFEL construction and its commission are reported here.

  8. PAL-XFEL cavity beam position monitor pick-up design and beam test

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Sojeong, E-mail: sojung8681@postech.ac.kr; Park, Young Jung; Kim, Changbum; Kim, Seung Hwan; Shin, Dong Cheol; Han, Jang-Hui; Ko, In Soo

    2016-08-11

    As an X-ray Free Electron Laser, PAL-XFEL is about to start beam commissioning. X-band cavity beam position monitor (BPM) is used in the PAL-XFEL undulator beam line. Prototypes of cavity BPM pick-up were designed and fabricated to test the RF characteristics. Also, the beam test of a cavity BPM pick-up was done in the Injector Test Facility (ITF). In the beam test, the raw signal properties of the cavity BPM pick-up were measured at a 200 pC bunch charge. According to the RF test and beam test results, the prototype cavity BPM pick-up design was confirmed to meet the requirements of the PAL-XFEL cavity BPM system.

  9. Mechanism of the Dual Activities of Human CYP17A1 and Binding to Anti-Prostate Cancer Drug Abiraterone Revealed by a Novel V366M Mutation Causing 17,20 Lyase Deficiency

    Directory of Open Access Journals (Sweden)

    Mónica Fernández-Cancio

    2018-04-01

    Full Text Available The CYP17A1 gene regulates sex steroid biosynthesis in humans through 17α-hydroxylase/17,20 lyase activities and is a target of anti-prostate cancer drug abiraterone. In a 46, XY patient with female external genitalia, together with a loss of function mutation S441P, we identified a novel missense mutation V366M at the catalytic center of CYP17A1 which preferentially impaired 17,20 lyase activity. Kinetic experiments with bacterially expressed proteins revealed that V366M mutant enzyme can bind and metabolize pregnenolone to 17OH-pregnenolone, but 17OH-pregnenolone binding and conversion to dehydroepiandrosterone (DHEA was impaired, explaining the patient’s steroid profile. Abiraterone could not bind and inhibit the 17α-hydroxylase activity of the CYP17A1-V366M mutant. Molecular dynamics (MD simulations showed that V366M creates a “one-way valve” and suggests a mechanism for dual activities of human CYP17A1 where, after the conversion of pregnenolone to 17OH-pregnenolone, the product exits the active site and re-enters for conversion to dehydroepiandrosterone. The V366M mutant also explained the effectiveness of the anti-prostate cancer drug abiraterone as a potent inhibitor of CYP17A1 by binding tightly at the active site in the WT enzyme. The V366M is the first human mutation to be described at the active site of CYP17A1 that causes isolated 17,20 lyase deficiency. Knowledge about the specificity of CYP17A1 activities is of importance for the development of treatments for polycystic ovary syndrome and inhibitors for prostate cancer therapy.

  10. Preparation of 1D Hierarchical Material Mesosilica/Pal Composite and Its Performance in the Adsorption of Methyl Orange

    Directory of Open Access Journals (Sweden)

    Mei Wu

    2018-01-01

    Full Text Available This paper highlights the synthesis of a one-dimensional (1D hierarchical material mesosilica/palygorskite (Pal composite and evaluates its adsorption performance for anionic dye methyl orange (MO in comparison with Pal and Mobile crystalline material-41 (MCM-41. The Mesosilica/Pal composite is consisted of mesosilica coated Pal nanorods and prepared through a dual template approach using cetyltrimethyl ammonium bromide (CTAB and Pal as soft and hard templates, respectively. The composition and structure of the resultant material was characterized by a scanning electron microscope (SEM, transmissionelectron microscopy (TEM, N2 adsorption-desorption analysis, small-angle X-Ray powder diffraction (XRD, and zeta potential measurement. Adsorption experiments were carried out with different absorbents at different contact times and pH levels. Compared with Pal and MCM-41, the mesosilica/Pal composite exhibited the best efficiency for MO adsorption. Its adsorption ratio is as high as 70.4%. Its adsorption equilibrium time is as short as 30 min. Results testify that the MO retention is promoted for the micro-mesoporous hierarchical structure and positive surface charge electrostatic interactions of the mesosilica/Pal composite. The regenerability of the mesosilica/Pal composite absorbent was also assessed. 1D morphology makes it facile to separate from aqueous solutions. It can be effortlessly recovered and reused for up to nine cycles.

  11. Cloning and Sequence Analysis of Vibrio halioticoli Genes Encoding Three Types of Polyguluronate Lyase.

    Science.gov (United States)

    Sugimura; Sawabe; Ezura

    2000-01-01

    The alginate lyase-coding genes of Vibrio halioticoli IAM 14596(T), which was isolated from the gut of the abalone Haliotis discus hannai, were cloned using plasmid vector pUC 18, and expressed in Escherichia coli. Three alginate lyase-positive clones, pVHB, pVHC, and pVHE, were obtained, and all clones expressed the enzyme activity specific for polyguluronate. Three genes, alyVG1, alyVG2, and alyVG3, encoding polyguluronate lyase were sequenced: alyVG1 from pVHB was composed of a 1056-bp open reading frame (ORF) encoding 352 amino acid residues; alyVG2 gene from pVHC was composed of a 993-bp ORF encoding 331 amino acid residues; and alyVG3 gene from pVHE was composed of a 705-bp ORF encoding 235 amino acid residues. Comparison of nucleotide and deduced amino acid sequences among AlyVG1, AlyVG2, and AlyVG3 revealed low homologies. The identity value between AlyVG1 and AlyVG2 was 18.7%, and that between AlyVG2 and AlyVG3 was 17.0%. A higher identity value (26.0%) was observed between AlyVG1 and AlyVG3. Sequence comparison among known polyguluronate lyases including AlyVG1, AlyVG2, and AlyVG3 also did not reveal an identical region in these sequences. However, AlyVG1 showed the highest identity value (36.2%) and the highest similarity (73.3%) to AlyA from Klebsiella pneumoniae. A consensus region comprising nine amino acid (YFKAGXYXQ) in the carboxy-terminal region previously reported by Mallisard and colleagues was observed only in AlyVG1 and AlyVG2.

  12. Quantitation of heparosan with heparin lyase III and spectrophotometry.

    Science.gov (United States)

    Huang, Haichan; Zhao, Yingying; Lv, Shencong; Zhong, Weihong; Zhang, Fuming; Linhardt, Robert J

    2014-02-15

    Heparosan is Escherichia coli K5 capsule polysaccharide, which is the key precursor for preparing bioengineered heparin. A rapid and effective quantitative method for detecting heparosan is important in the large-scale production of heparosan. Heparin lyase III (Hep III) effectively catalyzes the heparosan depolymerization, forming unsaturated disaccharides that are measurable using a spectrophotometer at 232 nm. We report a new method for the quantitative detection of heparosan with heparin lyase III and spectrophotometry that is safer and more specific than the traditional carbazole assay. In an optimized detection system, heparosan at a minimum concentration of 0.60 g/L in fermentation broth can be detected. Copyright © 2013 Elsevier Inc. All rights reserved.

  13. High-performance liquid chromatography-fluorescence assay of pyruvic acid to determine cysteine conjugate beta-lyase activity : application to S-1,2-dichlorovinyl-L-cysteine and S-2-benzothiazolyl-L-cysteine

    NARCIS (Netherlands)

    Stijntjes, G.J.; te Koppele, J.M.; Vermeulen, N P

    1992-01-01

    An HPLC-fluorescence assay has been developed for the determination of the activity of rat renal cytosolic cysteine conjugate beta-lyase. The method is based on isocratic HPLC separation and fluorescence detection of pyruvic acid, derivatized with o-phenylenediamine (OPD), and is shown to be rapid,

  14. Exploration of swapping enzymatic function between two proteins: A simulation study of chorismate mutase and isochorismate pyruvate lyase

    Science.gov (United States)

    Choutko, Alexandra; Eichenberger, Andreas P; Gunsteren, Wilfred F; Dolenc, Jožica

    2013-01-01

    The enzyme chorismate mutase EcCM from Escherichia coli catalyzes one of the few pericyclic reactions in biology, the transformation of chorismate to prephenate. The isochorismate pyruvate lyase PchB from Pseudomonas aeroginosa catalyzes another pericyclic reaction, the isochorismate to salicylate transformation. Interestingly, PchB possesses weak chorismate mutase activity as well thus being able to catalyze two distinct pericyclic reactions in a single active site. EcCM and PchB possess very similar folds, despite their low sequence identity. Using molecular dynamics simulations of four combinations of the two enzymes (EcCM and PchB) with the two substrates (chorismate and isochorismate) we show that the electrostatic field due to EcCM at atoms of chorismate favors the chorismate to prephenate transition and that, analogously, the electrostatic field due to PchB at atoms of isochorismate favors the isochorismate to salicylate transition. The largest differences between EcCM and PchB in electrostatic field strengths at atoms of the substrates are found to be due to residue side chains at distances between 0.6 and 0.8 nm from particular substrate atoms. Both enzymes tend to bring their non-native substrate in the same conformation as their native substrate. EcCM and to a lower extent PchB fail in influencing the forces on and conformations of the substrate such as to favor the other chemical reaction (isochorismate pyruvate lyase activity for EcCM and chorismate mutase activity for PchB). These observations might explain the difficulty of engineering isochorismate pyruvate lyase activity in EcCM by solely mutating active site residues. PMID:23595942

  15. Exploration of swapping enzymatic function between two proteins: a simulation study of chorismate mutase and isochorismate pyruvate lyase.

    Science.gov (United States)

    Choutko, Alexandra; Eichenberger, Andreas P; van Gunsteren, Wilfred F; Dolenc, Jožica

    2013-06-01

    The enzyme chorismate mutase EcCM from Escherichia coli catalyzes one of the few pericyclic reactions in biology, the transformation of chorismate to prephenate. The isochorismate pyruvate lyase PchB from Pseudomonas aeroginosa catalyzes another pericyclic reaction, the isochorismate to salicylate transformation. Interestingly, PchB possesses weak chorismate mutase activity as well thus being able to catalyze two distinct pericyclic reactions in a single active site. EcCM and PchB possess very similar folds, despite their low sequence identity. Using molecular dynamics simulations of four combinations of the two enzymes (EcCM and PchB) with the two substrates (chorismate and isochorismate) we show that the electrostatic field due to EcCM at atoms of chorismate favors the chorismate to prephenate transition and that, analogously, the electrostatic field due to PchB at atoms of isochorismate favors the isochorismate to salicylate transition. The largest differences between EcCM and PchB in electrostatic field strengths at atoms of the substrates are found to be due to residue side chains at distances between 0.6 and 0.8 nm from particular substrate atoms. Both enzymes tend to bring their non-native substrate in the same conformation as their native substrate. EcCM and to a lower extent PchB fail in influencing the forces on and conformations of the substrate such as to favor the other chemical reaction (isochorismate pyruvate lyase activity for EcCM and chorismate mutase activity for PchB). These observations might explain the difficulty of engineering isochorismate pyruvate lyase activity in EcCM by solely mutating active site residues. © 2013 The Protein Society.

  16. Polar localization of Escherichia coli chemoreceptors requires an intact Tol–Pal complex

    Science.gov (United States)

    Santos, Thiago M. A.; Lin, Ti-Yu; Rajendran, Madhusudan; Anderson, Samantha M.; Weibel, Douglas B.

    2014-01-01

    Summary Subcellular biomolecular localization is critical for the metabolic and structural properties of the cell. The functional implications of the spatiotemporal distribution of protein complexes during the bacterial cell cycle have long been acknowledged; however, the molecular mechanisms for generating and maintaining their dynamic localization in bacteria are not completely understood. Here we demonstrate that the trans-envelope Tol–Pal complex, a widely conserved component of the cell envelope of Gram-negative bacteria, is required to maintain the polar positioning of chemoreceptor clusters in Escherichia coli. Localization of the chemoreceptors was independent of phospholipid composition of the membrane and the curvature of the cell wall. Instead, our data indicate that chemoreceptors interact with components of the Tol–Pal complex and that this interaction is required to polarly localize chemoreceptor clusters. We found that disruption of the Tol–Pal complex perturbs the polar localization of chemoreceptors, alters cell motility, and affects chemotaxis. We propose that the E. coli Tol–Pal complex restricts mobility of the chemoreceptor clusters at the cell poles and may be involved in regulatory mechanisms that co-ordinate cell division and segregation of the chemosensory machinery. PMID:24720726

  17. Catalytically important amino-acid residues of abalone alginate lyase HdAly assessed by site-directed mutagenesis

    OpenAIRE

    Yamamoto, Sayo; Sahara, Takehiko; Sato, Daisuke; Kawasaki, Kosei; Ohgiya, Satoru; Inoue, Akira; Ojima, Takao

    2008-01-01

    Alginate lyase is an enzyme that degrades alginate chains via β-elimination and has been used for the production of alginate oligosaccharides and protoplasts from brown algae. Previously, we deduced the amino-acid sequence of an abalone alginate lyase, HdAly, from its cDNA sequence and, through multiple amino-acid sequence alignment, found that several basic amino-acid residues were highly conserved among the polysaccharide-lyase family 14 (PL-14) enzymes including HdAly. In the present study...

  18. Light and Fungal Elicitor Induce 3-Deoxy-d-arabino-Heptulosonate 7-Phosphate Synthase mRNA in Suspension Cultured Cells of Parsley (Petroselinum crispum L.) 1

    Science.gov (United States)

    Henstrand, John M.; McCue, Kent F.; Brink, Kent; Handa, Avtar K.; Herrmann, Klaus M.; Conn, Eric E.

    1992-01-01

    Light and fungal elicitor induce mRNA encoding 3-deoxy-d-arabino-heptulosonate 7-phosphate (DAHP) synthase in suspension cultured cells of parsley (Petroselinum crispum L.). The kinetics and dose response of mRNA accumulation were similar for DAHP synthase and phenylalanine ammonia-lyase (PAL). Six micrograms of elicitor from Phytophthora megasperma f. glycinia gave a detectable induction within 1 hour. Induction of DAHP synthase and PAL mRNAs by light was transient, reaching maximal levels at 4 hours and returning to pretreatment levels after 24 hours. Our data suggest that either light or fungal elicitor transcriptionally activate DAHP synthase. A coordinate regulation for key enzymes in the synthesis of primary and secondary metabolites is indicated. ImagesFigure 1 PMID:16668708

  19. Secondary metabolism and antioxidants are involved in environmental adaptation and stress tolerance in lettuce.

    Science.gov (United States)

    Oh, Myung-Min; Trick, Harold N; Rajashekar, C B

    2009-01-30

    Lettuce (Lactuca sativa) plants grown in a protective environment, similar to in vitro conditions, were acclimated in a growth chamber and subjected to water stress to examine the activation of genes involved in secondary metabolism and biosynthesis of antioxidants. The expression of phenylalanine ammonia-lyase (PAL), gamma-tocopherol methyl transferase (gamma-TMT) and l-galactose dehydrogenase (l-GalDH) genes involved in the biosynthesis of phenolic compounds, alpha-tocopherol and ascorbic acid, respectively, were determined during plant adaptation. These genes were activated in tender plants, grown under protective conditions, when exposed to normal growing conditions in a growth chamber. A large increase in transcript level for PAL, a key gene in the phenylpropanoid pathway leading to the biosynthesis of a wide array of phenolics and flavonoids, was observed within 1h of exposure of tender plants to normal growing conditions. Plant growth, especially the roots, was retarded in tender plants when exposed to normal growing conditions. Furthermore, exposure of both protected and unprotected plants to water stress resulted in the activation of PAL. PAL inhibition by 2-aminoindan-2-phosphonic acid (AIP) rendered these plants more sensitive to chilling and heat shock treatments. These results suggest that activation of secondary metabolism as well as the antioxidative metabolism is an integral part of plant adaptation to normal growing conditions in lettuce plants.

  20. Ecosystem Pen Pals: Using Place-Based Marine Science and Culture to Connect Students

    Science.gov (United States)

    Wiener, Carlie S.; Matsumoto, Karen

    2014-01-01

    The marine environment provides a unique context for students to explore both natural and cultural connections. This paper reports preliminary findings on Ecosystem Pen Pals, an ocean literacy program for 4th and 5th graders focused on using a pen pal model for integrating traditional ecological knowledge into marine science. Surveys with…

  1. Melatonin treatment attenuates postharvest decay and maintains nutritional quality of strawberry fruits (Fragaria×anannasa cv. Selva) by enhancing GABA shunt activity.

    Science.gov (United States)

    Aghdam, Morteza Soleimani; Fard, Javad Rezapour

    2017-04-15

    Fresh strawberry fruits as perishable commodities have a short postharvest life and are prone to postharvest fungal decay. In this study, the impact of 0, 1, 10, 100 and 1000μmol/L melatonin on attenuating fungal decay and maintaining nutritional quality of strawberry fruits was investigated during storage at 4°C for 12days. Melatonin treatment at 100μmol/L triggered H 2 O 2 accumulation, which result from higher superoxide dismutase (SOD) activity, associated with lower catalase (CAT) and ascorbate peroxidase (APX) activities, leading to fruits with lower decay. Higher H 2 O 2 accumulation was concurrent with higher phenylalanine ammonia lyase (PAL) enzyme activity leading to higher total phenols and anthocyanins accumulation along with higher DPPH scavenging capacity. Also, strawberry fruits treated with melatonin exhibited higher γ-aminobutyric acid transaminase (GABA-T) enzyme activity which ensured sufficient ATP supplying leading to higher unsaturated/saturated fatty acids (unSFA/SFA) ratio. Copyright © 2016 Elsevier Ltd. All rights reserved.

  2. Design of thermostable rhamnogalacturonan lyase mutants from Bacillus licheniformis by combination of targeted single point mutations

    DEFF Research Database (Denmark)

    da Silva, Ines Isabel Cardoso Rodrigues; Jers, Carsten; Otten, Harm

    2014-01-01

    Rhamnogalacturonan I lyases (RGI lyases) (EC 4.2.2.-) catalyze cleavage of α-1,4 bonds between rhamnose and galacturonic acid in the backbone of pectins by β-elimination. In the present study, targeted improvement of the thermostability of a PL family 11 RGI lyase from Bacillus licheniformis (DSM......, were obtained due to additive stabilizing effects of single amino acid mutations (E434L, G55V, and G326E) compared to the wild type. The crystal structure of the B. licheniformis wild-type RGI lyase was also determined; the structural analysis corroborated that especially mutation of charged amino...

  3. Thiamine primed defense provides reliable alternative to systemic fungicide carbendazim against sheath blight disease in rice (Oryza sativa L.).

    Science.gov (United States)

    Bahuguna, Rajeev Nayan; Joshi, Rohit; Shukla, Alok; Pandey, Mayank; Kumar, J

    2012-08-01

    A novel pathogen defense strategy by thiamine priming was evaluated for its efficacy against sheath blight pathogen, Rhizoctonia solani AG-1A, of rice and compared with that of systemic fungicide, carbendazim (BCM). Seeds of semidwarf, high yielding, basmati rice variety Vasumati were treated with thiamine (50 mM) and BCM (4 mM). The pot cultured plants were challenge inoculated with R. solani after 40 days of sowing and effect of thiamine and BCM on rice growth and yield traits was examined. Higher hydrogen peroxide content, total phenolics accumulation, phenylalanine ammonia lyase (PAL) activity and superoxide dismutase (SOD) activity under thiamine treatment displayed elevated level of systemic resistance, which was further augmented under challenging pathogen infection. High transcript level of phenylalanine ammonia lyase (PAL) and manganese superoxide dismutase (MnSOD) validated mode of thiamine primed defense. Though minimum disease severity was observed under BCM treatment, thiamine produced comparable results, with 18.12 per cent lower efficacy. Along with fortifying defense components and minor influence on photosynthetic pigments and nitrate reductase (NR) activity, thiamine treatment significantly reduced pathogen-induced loss in photosynthesis, stomatal conductance, chlorophyll fluorescence, NR activity and NR transcript level. Physiological traits affected under pathogen infection were found signatory for characterizing plant's response under disease and were detectable at early stage of infection. These findings provide a novel paradigm for developing alternative, environmentally safe strategies to control plant diseases. Copyright © 2012 Elsevier Masson SAS. All rights reserved.

  4. Renal cysteine conjugate C-S lyase mediated toxicity of halogenated alkenes in primary cultures of human and rat proximal tubular cells.

    Science.gov (United States)

    McGoldrick, Trevor A; Lock, Edward A; Rodilla, Vicente; Hawksworth, Gabrielle M

    2003-07-01

    Proximal tubular cells from human (HPT) and rat (RPT) kidneys were isolated, grown to confluence and incubated with S-(1,2-dichlorovinyl)- l-cysteine (DCVC), S-(1,2,2-trichlorovinyl)- l-cysteine (TCVC), S-(1,1,2,2-tetrafluoroethyl)- l-cysteine (TFEC) and S-(2-chloro-1,1-difluorethyl)- l-cysteine (CDFEC), the cysteine conjugates of nephrotoxicants. The cultures were exposed to the conjugates for 12, 24 and 48 h and the toxicity determined using the MTT assay. All four conjugates caused dose-dependent toxicity to RPT cells over the range 50-1,000 microM, the order of toxicity being DCVC>TCVC>TFEC=CDFEC. The inclusion of aminooxyacetic acid (AOAA; 250 microM), an inhibitor of pyridoxal phosphate-dependent enzymes such as C-S lyase, afforded protection, indicating that C-S lyase has a role in the bioactivation of these conjugates. In HPT cultures only DCVC caused significant time- and dose-dependent toxicity. Exposure to DCVC (500 microM) for 48 h decreased cell viability to 7% of control cell values, whereas co-incubation of DCVC (500 microM) with AOAA (250 microM) resulted in cell viability of 71%. Human cultures were also exposed to S-(1,2-dichlorovinyl)-glutathione (DCVG). DCVG was toxic to HPT cells, but the onset of toxicity was delayed compared with the corresponding cysteine conjugate. AOAA afforded almost complete protection from DCVG toxicity. Acivicin (250 microM), an inhibitor of gamma-glutamyl transferase (gamma-GT), partially protected against DCVG (500 microM)-induced toxicity at 48 h (5% viability and 53% viability in the absence and presence of acivicin, respectively). These results suggest that DCVG requires processing by gamma-GT prior to bioactivation by C-S lyase in HPT cells. The activity of C-S lyase, using TFEC as a substrate, and glutamine transaminase K (GTK) was measured in rat and human cells with time in culture. C-S lyase activity in RPT and HPT cells decreased to approximately 30% of fresh cell values by the time the cells reached

  5. The effect of peer assisted learning (PAL) on anatomic competencies of prospective student’s biology teachers

    Science.gov (United States)

    Diana, S.

    2018-05-01

    Research about the effect of Peer Assisted Learning (PAL) strategy on Plant Anatomy Course, which aims to improve anatomic competencies of prospective student’s biology teacher, has been done. This study used a quasi-experimental pre-test post-test control group design. The draft includes a group of students who were given a pre-test which is then followed by observing the PAL treatment process and post-test. The other students group (control) was given the pre-test and post-test only. The PAL program began with a discussion between the lecturer and the tutor about the pretest results and then discussion between the tutors and their tutees in the class about the responses items. After that, all students were assigned to answer a set of response items, and then did the posttest. The results showed that the PAL strategy can increase student’s anatomic literacy significantly and can increase student’s anatomic lab skills no significantly. Thus the PAL strategy implementation has a potential to improve student’s anatomic competencies. The generally students weaknesses were they lack practice in interpreting of research results in the graphs form and rarely shared about anatomic lab skills. All students respond positively to the PAL strategy.

  6. Monoclonal antibody PAL-E specific for endothelium

    NARCIS (Netherlands)

    Schlingemann, R. O.; Dingjan, G. M.; Emeis, J. J.; Blok, J.; Warnaar, S. O.; Ruiter, D. J.

    1985-01-01

    A monoclonal antibody, PAL-E, is described that is specific for endothelial cells. The monoclonal antibody, an IgG2a, markedly stains endothelium of capillaries, medium-sized and small veins, and venules in frozen sections of human and some animal tissues tested. It reacts not at all or only weakly

  7. Salicylic acid induces vanillin synthesis through the phospholipid signaling pathway in Capsicum chinense cell cultures.

    Science.gov (United States)

    Rodas-Junco, Beatriz A; Cab-Guillén, Yahaira; Muñoz-Sánchez, J Armando; Vázquez-Flota, Felipe; Monforte-González, Miriam; Hernández-Sotomayor, S M Teresa

    2013-10-01

    Signal transduction via phospholipids is mediated by phospholipases such as phospholipase C (PLC) and D (PLD), which catalyze hydrolysis of plasma membrane structural phospholipids. Phospholipid signaling is also involved in plant responses to phytohormones such as salicylic acid (SA). The relationships between phospholipid signaling, SA, and secondary metabolism are not fully understood. Using a Capsicum chinense cell suspension as a model, we evaluated whether phospholipid signaling modulates SA-induced vanillin production through the activation of phenylalanine ammonia lyase (PAL), a key enzyme in the biosynthetic pathway. Salicylic acid was found to elicit PAL activity and consequently vanillin production, which was diminished or reversed upon exposure to the phosphoinositide-phospholipase C (PI-PLC) signaling inhibitors neomycin and U73122. Exposure to the phosphatidic acid inhibitor 1-butanol altered PLD activity and prevented SA-induced vanillin production. Our results suggest that PLC and PLD-generated secondary messengers may be modulating SA-induced vanillin production through the activation of key biosynthetic pathway enzymes.

  8. The federated ontology of the pal project interfacing ontologies and integrating time-dependent data

    NARCIS (Netherlands)

    Krieger, H.U.; Peters, R.; Kiefer, B.; Van Bekkum, M.A.; Kaptein, F.; Neerincx, M.A.

    2016-01-01

    This paper describes ongoing work carried out in the European project PAL which will support childre in their diabetes self-management as well as assist health professionals and parents involved in the diabete regimen of the child. Here, we will focus on the construction of the PAL ontology which

  9. Hematopoietic Sphingosine 1-Phosphate Lyase Deficiency Decreases Atherosclerotic Lesion Development in LDL-Receptor Deficient Mice

    NARCIS (Netherlands)

    Bot, Martine; Van Veldhoven, Paul P.; de Jager, Saskia C. A.; Johnson, Jason; Nijstad, Niels; Van Santbrink, Peter J.; Westra, Marijke M.; Van Der Hoeven, Gerd; Gijbels, Marion J.; Mueller-Tidow, Carsten; Varga, Georg; Tietge, Uwe J. F.; Kuiper, Johan; Van Berkel, Theo J. C.; Nofer, Jerzy-Roch; Bot, Ilze; Biessen, Erik A. L.

    2013-01-01

    Aims: Altered sphingosine 1-phosphate (S1P) homeostasis and signaling is implicated in various inflammatory diseases including atherosclerosis. As S1P levels are tightly controlled by S1P lyase, we investigated the impact of hematopoietic S1P lyase (Sgpl1(-/-)) deficiency on leukocyte subsets

  10. Hematopoietic sphingosine 1-phosphate lyase deficiency decreases atherosclerotic lesion development in LDL-receptor deficient mice

    NARCIS (Netherlands)

    Bot, Martine; van Veldhoven, Paul P.; de Jager, Saskia C. A.; Johnson, Jason; Nijstad, Niels; van Santbrink, Peter J.; Westra, Marijke M.; van der Hoeven, Gerd; Gijbels, Marion J.; Müller-Tidow, Carsten; Varga, Georg; Tietge, Uwe J. F.; Kuiper, Johan; van Berkel, Theo J. C.; Nofer, Jerzy-Roch; Bot, Ilze; Biessen, Erik A. L.

    2013-01-01

    Altered sphingosine 1-phosphate (S1P) homeostasis and signaling is implicated in various inflammatory diseases including atherosclerosis. As S1P levels are tightly controlled by S1P lyase, we investigated the impact of hematopoietic S1P lyase (Sgpl1(-/-)) deficiency on leukocyte subsets relevant to

  11. Development of new S-band SLED for PAL-XFEL Linac

    Energy Technology Data Exchange (ETDEWEB)

    Joo, Youngdo; Park, Yongjung; Heo, Hoon; Heo, Jinyul; Park, Sung-Soo; Kim, Sang-Hee; Kim, Kwang-Hoon; Kang, Heung-Sik [Pohang Accelerator Laboratory, Pohang, Gyeongbuk 37673 (Korea, Republic of); Lee, Heung-Soo, E-mail: lhs@postech.ac.kr [Pohang Accelerator Laboratory, Pohang, Gyeongbuk 37673 (Korea, Republic of); Noh, Sungju; Oh, Kyoungmin [VitzroTech, Ansan, Gyeonggi 15603 (Korea, Republic of)

    2017-01-21

    In order to achieve beam acceleration to the beam energy of 10 GeV at the end of its 716 m-long linear accelerator (Linac), the Pohang Accelerator Laboratory X-ray Free Electron Laser (PAL-XFEL) is going to operate the Stanford Linear Accelerator Energy Doubler (SLED) at the maximum klystron output peak power of 80 MW, with a pulse length of 4 μs, and at a repetition rate of 60 Hz. The original SLED that had been used in Pohang Light Source-II (PLS-II) can no longer sustain such a high-power operation because excessive radiation caused by RF breakdown has been frequently detected even at the lower klystron peak power during the PLS-II operation. Therefore, a new SLED is designed by modifying both the 3-dB power hybrid and the waveguide-cavity coupling structure of the original SLED where the excessive radiation has been mainly detected. The finite-difference time-domain (FDTD) simulation in the CST Microwave Studio shows that the new SLED has a peak electric field and a surface current lower than those of the original SLED at the same level of the RF input peak power, which would secure stable high-power operation. All of the 42 SLEDs in the PAL-XFEL Linac are newly fabricated and installed. During the RF conditioning of the PAL-XFEL Linac, no significant vacuum and radiation issue was found in the new SLEDs. Finally, the accelerated electron beam energy of 10 GeV obtained at the end of the PAL-XFEL Linac verified that the RF performance of the new SLED is stable.

  12. Development of new S-band SLED for PAL-XFEL Linac

    Science.gov (United States)

    Joo, Youngdo; Park, Yongjung; Heo, Hoon; Heo, Jinyul; Park, Sung-Soo; Kim, Sang-Hee; Kim, Kwang-Hoon; Kang, Heung-Sik; Lee, Heung-Soo; Noh, Sungju; Oh, Kyoungmin

    2017-01-01

    In order to achieve beam acceleration to the beam energy of 10 GeV at the end of its 716 m-long linear accelerator (Linac), the Pohang Accelerator Laboratory X-ray Free Electron Laser (PAL-XFEL) is going to operate the Stanford Linear Accelerator Energy Doubler (SLED) at the maximum klystron output peak power of 80 MW, with a pulse length of 4 μs, and at a repetition rate of 60 Hz. The original SLED that had been used in Pohang Light Source-II (PLS-II) can no longer sustain such a high-power operation because excessive radiation caused by RF breakdown has been frequently detected even at the lower klystron peak power during the PLS-II operation. Therefore, a new SLED is designed by modifying both the 3-dB power hybrid and the waveguide-cavity coupling structure of the original SLED where the excessive radiation has been mainly detected. The finite-difference time-domain (FDTD) simulation in the CST Microwave Studio shows that the new SLED has a peak electric field and a surface current lower than those of the original SLED at the same level of the RF input peak power, which would secure stable high-power operation. All of the 42 SLEDs in the PAL-XFEL Linac are newly fabricated and installed. During the RF conditioning of the PAL-XFEL Linac, no significant vacuum and radiation issue was found in the new SLEDs. Finally, the accelerated electron beam energy of 10 GeV obtained at the end of the PAL-XFEL Linac verified that the RF performance of the new SLED is stable.

  13. Endothelial cell marker PAL-E reactivity in brain tumor, developing brain, and brain disease

    NARCIS (Netherlands)

    Leenstra, S.; Troost, D.; Das, P. K.; Claessen, N.; Becker, A. E.; Bosch, D. A.

    1993-01-01

    The endothelial cell marker PAL-E is not reactive to vessels in the normal brain. The present study concerns the PAL-E reactivity in brain tumors in contrast to normal brain and nonneoplastic brain disease. A total of 122 specimens were examined: brain tumors (n = 94), nonneoplastic brain disease (n

  14. Active tuberculosis patients have high levels of IgA anti-alpha-crystallin and isocitrate lyase proteins.

    Science.gov (United States)

    Talavera-Paulín, M; García-Morales, L; Ruíz-Sánchez, B P; Caamal-Ley, Á D; Hernández-Solis, A; Ramírez-Casanova, E; Cicero-Sabido, R; Espitia, C; Helguera-Repetto, C; González-Y-Merchand, J A; Flores-Mejía, R; Estrada-Parra, S; Estrada-García, I; Chacón-Salinas, R; Wong-Baeza, I; Serafín-López, J

    2016-12-01

    Mexico City, Mexico. To identify proteins synthetised by Mycobacterium tuberculosis in hypoxic culture, which resemble more closely a granuloma environment than aerobic culture, and to determine if they are recognised by antibodies from patients with active pulmonary tuberculosis (PTB). Soluble extracts from M. tuberculosis H37Rv cultured under aerobic or hypoxic conditions were analysed using two-dimensional polyacrylamide gel electrophoresis, and proteins over-expressed under hypoxia were identified by mass spectrometry. The presence of immunoglobulin (Ig) G, IgA and IgM antibodies against these proteins was determined in the serum of 42 patients with active PTB and 42 healthy controls. We selected three M. tuberculosis H37Rv proteins (alpha-crystallin protein [Acr, Rv2031c], universal stress protein Rv2623 and isocitrate lyase [ICL, RV0467]) that were over-expressed under hypoxia. Titres of anti-Acr and anti-ICL IgA antibodies were higher in patients than in healthy controls, with an area under the receiver operating characteristic curve of 0.71 for anti-ICL IgA antibodies. ICL could be used in combination with other M. tuberculosis antigens to improve the sensitivity and specificity of current serological TB diagnostic methods.

  15. Hematopoietic Sphingosine 1-Phosphate Lyase Deficiency Decreases Atherosclerotic Lesion Development in LDL-Receptor Deficient Mice

    NARCIS (Netherlands)

    Bot, M.; Veldhoven, van P.P.; Jager, de S.C.; Johnson, J.; Nijstad, N.; van, Santbrink P.J.; Westra, M.M.; Hoeven, van der G.; Gijbels, M.J.; Muller-Tidow, C.; Varga, G.; Tietge, U.J.; Kuiper, J.; Berkel, van T.J.; Nofer, J.R.; Bot, I.; Biessen, E.A.

    2013-01-01

    Abstract Aims Altered sphingosine 1-phosphate (S1P) homeostasis and signaling is implicated in various inflammatory diseases including atherosclerosis. As S1P levels are tightly controlled by S1P lyase, we investigated the impact of hematopoietic S1P lyase (Sgpl1−/−) deficiency on leukocyte

  16. Study design and protocol for a mixed methods evaluation of an intervention to reduce and break up sitting time in primary school classrooms in the UK: The CLASS PAL (Physically Active Learning) Programme.

    Science.gov (United States)

    Routen, Ash C; Biddle, Stuart J H; Bodicoat, Danielle H; Cale, Lorraine; Clemes, Stacy; Edwardson, Charlotte L; Glazebrook, Cris; Harrington, Deirdre M; Khunti, Kamlesh; Pearson, Natalie; Salmon, Jo; Sherar, Lauren B

    2017-11-08

    Children engage in a high volume of sitting in school, particularly in the classroom. A number of strategies, such as physically active lessons (termed movement integration (MI)), have been developed to integrate physical activity into this learning environment; however, no single approach is likely to meet the needs of all pupils and teachers. This protocol outlines an implementation study of a primary school-based MI intervention: CLASS PAL (Physically Active Learning) programme. This study aims to (A) determine the degree of implementation of CLASS PAL, (B) identify processes by which teachers and schools implement CLASS PAL and (C) investigate individual (pupil and teacher) level and school-level characteristics associated with implementation of CLASS PAL. The intervention will provide teachers with a professional development workshop and a bespoke teaching resources website. The study will use a single group before-and-after design, strengthened by multiple interim measurements. Six state-funded primary schools will be recruited within Leicestershire, UK.Evaluation data will be collected prior to implementation and at four discrete time points during implementation: At measurement 0 (October 2016), school, teacher and pupil characteristics will be collected. At measurements 0 and 3 (June-July 2017), accelerometry, cognitive functioning, self-reported sitting and classroom engagement data will be collected. At measurements 1(December 2016-March 2017) and 3 , teacher interviews (also at measurement 4; September-October 2017) and pupil focus groups will be conducted, and at measurements 1 and 2 (April-May 2017), classroom observations. Implementation will be captured through website analytics and ongoing teacher completed logs. Ethical approval was obtained through the Loughborough University Human Participants Ethics Sub-Committee (Reference number: R16-P115). Findings will be disseminated via practitioner and/or research journals and to relevant regional and

  17. Sequence analysis and overexpression of a pectin lyase gene (pel1) from Aspergillus oryzae KBN616.

    Science.gov (United States)

    Kitamoto, N; Yoshino-Yasuda, S; Ohmiya, K; Tsukagoshi, N

    2001-01-01

    A gene (pel1) encoding pectin lyase (Pel1) was isolated from a shoyu koji mold, Aspergillus oryzae KBN616, and characterized. The structural gene comprised 1,196 bp with a single intron. The ORF encoded 381 amino acids with a signal peptide of 20 amino acids. The deduced amino acid sequence showed high similarity to those of Aspergillus niger pectin lyases and Glomerella cingulata PnlA. The pel1 gene was successfully overexpressed under the promoter of the A. oryzae TEF1 gene. The molecular mass of the recombinant pectin lyase substantially coincided with that calculated based on nucleotide sequence.

  18. PowerPal: power up your creek

    International Nuclear Information System (INIS)

    2006-01-01

    PowerPal micro-hydro equipment was developed in Vietnam by Asian Phoenix Resources Ltd. of Victoria, BC; a company who aims to help disadvantaged people in less developed countries by making renewable energy devices available to them. The idea for the micro-hydro system was conceived after mineral exploration parties in Vietnam noted that large numbers of poor quality propeller turbines were being used to supply AC power to individual homes. A plan was developed to improve the quality and reliability of low head turbines with better castings, magnets, superior bearings and electronic load controllers. Funding for the program was initially difficult to obtain. However, PowerPal micro-hydro equipment eventually came to the attention of the World Bank as well as other government and non-governmental organizations. Financing for the purchase and installation of larger numbers of units has now been achieved, and missionary groups in various parts of the world have ordered units. European markets have also opened up, and distributors of the micro-hydro equipment have been appointed in various countries. Sales are now strong in Central America, South America, the Caribbean, Australasia, Oceania, Africa and Europe. It was concluded that the surge of interest has coincided with a steep upturn in oil prices. 8 figs

  19. Effect of temperature on enzymatic and physiological factors related to chilling injury in carambola fruit (Averrhoa carambola L.).

    Science.gov (United States)

    Pérez-Tello, G O; Silva-Espinoza, B A; Vargas-Arispuro, I; Briceño-Torres, B O; Martinez-Tellez, M A

    2001-10-05

    Three groups of carambola fruits (Averrhoa carambola L.) were stored at 2 and 10 degrees C (85-90% relative humidity). The major physicochemical, physiological, and enzymatic responses of fruit were measured in each group over a 30-day period: chilling injury index (CII), decay (%), intracuticular waxes, cuticle permeability, pulp firmness, weight loss, sucrose, fructose and glucose contents, ion electrolyte leakage in pulp (%), ethylene and carbon dioxide production rates, and the activities of peroxidase (POD), polyphenol oxidase (PPO), and phenylalanine ammonia-lyase (PAL) enzymes. CII values were statistically different at 2 and 10 degrees C, showing high significance with respect to sucrose content and weight loss (P < 0.05). Chilling injury included darkened ribs and skin desiccation. According to the CI symptom development, a possible relationship of POD and PPO activities was found at 2 degrees C. A significant sucrose content increase was observed at 10 degrees C. CI symptoms were associated with POD and PAL activities. Copyright 2001 Academic Press.

  20. Targeting sphingosine-1-phosphate lyase as an anabolic therapy for bone loss.

    Science.gov (United States)

    Weske, Sarah; Vaidya, Mithila; Reese, Alina; von Wnuck Lipinski, Karin; Keul, Petra; Bayer, Julia K; Fischer, Jens W; Flögel, Ulrich; Nelsen, Jens; Epple, Matthias; Scatena, Marta; Schwedhelm, Edzard; Dörr, Marcus; Völzke, Henry; Moritz, Eileen; Hannemann, Anke; Rauch, Bernhard H; Gräler, Markus H; Heusch, Gerd; Levkau, Bodo

    2018-05-01

    Sphingosine-1-phosphate (S1P) signaling influences bone metabolism, but its therapeutic potential in bone disorders has remained unexplored. We show that raising S1P levels in adult mice through conditionally deleting or pharmacologically inhibiting S1P lyase, the sole enzyme responsible for irreversibly degrading S1P, markedly increased bone formation, mass and strength and substantially decreased white adipose tissue. S1P signaling through S1P 2 potently stimulated osteoblastogenesis at the expense of adipogenesis by inversely regulating osterix and PPAR-γ, and it simultaneously inhibited osteoclastogenesis by inducing osteoprotegerin through newly discovered p38-GSK3β-β-catenin and WNT5A-LRP5 pathways. Accordingly, S1P 2 -deficient mice were osteopenic and obese. In ovariectomy-induced osteopenia, S1P lyase inhibition was as effective as intermittent parathyroid hormone (iPTH) treatment in increasing bone mass and was superior to iPTH in enhancing bone strength. Furthermore, lyase inhibition in mice successfully corrected severe genetic osteoporosis caused by osteoprotegerin deficiency. Human data from 4,091 participants of the SHIP-Trend population-based study revealed a positive association between serum levels of S1P and bone formation markers, but not resorption markers. Furthermore, serum S1P levels were positively associated with serum calcium , negatively with PTH , and curvilinearly with body mass index. Bone stiffness, as determined through quantitative ultrasound, was inversely related to levels of both S1P and the bone formation marker PINP, suggesting that S1P stimulates osteoanabolic activity to counteract decreasing bone quality. S1P-based drugs should be considered as a promising therapeutic avenue for the treatment of osteoporotic diseases.

  1. One NASA: Sharing Knowledge Through an Agency-wide Process Asset Library (PAL)

    Science.gov (United States)

    Truss, Baraka J.

    2006-01-01

    This poster session will cover the key purpose and components behind implementing the NASA PAL website. This session will present the current results, describing the process used to create the website, the current usage measure, and will demonstrate how NASA is truly becoming ONE. The target audience for the poster session includes those currently implementing the CMMI model and looking for PAL adoption techniques. To continue to be the leader in space, science and technology, NASA is using this agency-wide PAL to share knowledge, work products and lessons learned through this website. Many organizations have failed to recognize how the efforts of process improvement fit into overall organizational effort. However, NASA as an agency has adopted the benefits of process improvement by the creation of this website to foster communication between its ten centers. The poster session will cover the following, topics outlined below: 1) Website purpose; 2) Characteristics of the website; 3) User accounts status; 4) Website content size; and 5) Usage percentages.

  2. Abundance and genetic diversity of microbial polygalacturonase and pectate lyase in the sheep rumen ecosystem.

    Directory of Open Access Journals (Sweden)

    Peng Yuan

    Full Text Available Efficient degradation of pectin in the rumen is necessary for plant-based feed utilization. The objective of this study was to characterize the diversity, abundance, and functions of pectinases from microorganisms in the sheep rumen.A total of 103 unique fragments of polygalacturonase (PF00295 and pectate lyase (PF00544 and PF09492 genes were retrieved from microbial DNA in the rumen of a Small Tail Han sheep, and 66% of the sequences of these fragments had low identities (<65% with known sequences. Phylogenetic tree building separated the PF00295, PF00544, and PF09492 sequences into five, three, and three clades, respectively. Cellulolytic and noncellulolytic Butyrivibrio, Prevotella, and Fibrobacter species were the major sources of the pectinases. The two most abundant pectate lyase genes were cloned, and their protein products, expressed in Escherichia coli, were characterized. Both enzymes probably act extracellularly as their nucleotide sequences contained signal sequences, and they had optimal activities at the ruminal physiological temperature and complementary pH-dependent activity profiles.This study reveals the specificity, diversity, and abundance of pectinases in the rumen ecosystem and provides two additional ruminal pectinases for potential industrial use under physiological conditions.

  3. Biotechnology of humified materials obtained from vermicomposts ...

    African Journals Online (AJOL)

    PERSON

    2013-02-13

    Feb 13, 2013 ... Techniques for determining the structure of HS have been particularly useful for .... Step 6: Attempted introduction of the technologies into agro- industrial and .... rice plants stimulated several enzymatic mechanisms associated with the ... ammonia-lyase (PAL) were stimulated, resulting in increased proline ...

  4. Potential effects of plant growth promoting rhizobacteria ...

    African Journals Online (AJOL)

    Damping off caused by Sclerotium rolfsii on cowpea results in yield losses with serious socioeconomic implication. Induction of defense responses by plant growth promoting rhizobacteria (PGPR) is largely associated with the production of defense enzyme phenyl ammonia lyase (PAL) and oxidative enzymes like ...

  5. Phenolic metabolites in carnivorous plants: Inter-specific comparison and physiological studies.

    Science.gov (United States)

    Kováčik, Jozef; Klejdus, Bořivoj; Repčáková, Klára

    2012-03-01

    Despite intensive phytochemical research, data related to the accumulation of phenols in carnivorous plants include mainly qualitative reports. We have quantified phenolic metabolites in three species: Drosera capensis, Dionaea muscipula and Nepenthes anamensis in the "leaf" (assimilatory part) and the "trap" (digestive part). For comparison, commercial green tea was analysed. Phenylalanine ammonia-lyase (PAL) activities in Dionaea and Nepenthes were higher in the trap than in the leaf while the opposite was found in Drosera. Soluble phenols and majority of phenolic acids were mainly accumulated in the trap among species. Flavonoids were abundant in Drosera and Dionaea traps but not in Nepenthes. Phenolic acids were preferentially accumulated in a glycosidically-bound form and gallic acid was the main metabolite. Green tea contained more soluble phenols and phenolic acids but less quercetin. In vitro experiments with Drosera spathulata revealed that nitrogen deficiency enhances PAL activity, accumulation of phenols and sugars while PAL inhibitor (2-aminoindane-2-phosphonic acid) depleted phenols and some amino acids (but free phenylalanine and sugars were elevated). Possible explanations in physiological, biochemical and ecological context are discussed. Copyright © 2011 Elsevier Masson SAS. All rights reserved.

  6. Internet Safety and Malaysian Students: A Study of Female Undergraduates and Their Net Pals

    Directory of Open Access Journals (Sweden)

    Cheong Lee Mei, Dianne

    2007-12-01

    Full Text Available Malaysian female undergraduates are perceived as able to rationally evaluate and estimate risks towards Net pals. Friendship is secured online while deception and fraud are revealed offline. Based on what they have been informed, the female undergraduates may know their Net pals’ likes and dislikes but the invisible cyber veil does not allow them to penetrate the illusion and accurately evaluate a person’s character and personality. This study examined how Malaysian female undergraduates use the Internet, perceive risks in meeting Net pals and the safety awareness surrounding cyber friendship. Responses of 118 female undergraduates (43% from UiTM, 30% from UM, 27% from MMU to a survey indicated that Net pals were befriended through chat rooms and websites. More than half of the female undergraduates would have a face-to-face meeting with their Net pals without the knowledge of relevant parties and have done at least one potentially unsafe behaviour. If the extent and level of Internet safety awareness is known, the problem of Internet deception that might lead to potential social problems such as rape and sexual abuse can be effectively addressed.

  7. Biochemical responses during the pathogenesis of Sclerotium rolfsii ...

    African Journals Online (AJOL)

    Subhadfip Nandi

    a variety of novel proteins and secondary metabolites. This study aimed to examine the induction of different stress related enzymes like phenyl alanineammonia lyase (PAL), chitinase, β-1,3 glucanase, oxidative enzymes like peroxidases (POD), poly phenol oxidases (PPO) and phenolics after inoculation of Sclerotium ...

  8. Crystal structure and characterization of a novel L-serine ammonia-lyase from Rhizomucor miehei

    Energy Technology Data Exchange (ETDEWEB)

    Qin, Zhen [College of Food Science and Nutritional Engineering, Beijing Advanced Innovation Center of Food Nutrition and Human Health, China Agricultural University, Beijing 100083 (China); Yan, Qiaojuan [College of Engineering, China Agricultural University, Beijing 100083 (China); Ma, Qingjun [Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071 (China); Jiang, Zhengqiang, E-mail: zhqjiang@cau.edu.cn [College of Food Science and Nutritional Engineering, Beijing Advanced Innovation Center of Food Nutrition and Human Health, China Agricultural University, Beijing 100083 (China)

    2015-10-23

    L-serine ammonia-lyase, as a member of the β-family of pyridoxal-5′-phosphate (PLP) dependent enzymes, catalyzes the conversion of L-serine (L-threonine) to pyruvate (α-ketobutyrate) and ammonia. The crystal structure of L-serine ammonia-lyase from Rhizomucor miehei (RmSDH) was solved at 1.76 Å resolution by X-ray diffraction method. The overall structure of RmSDH had the characteristic β-family PLP dependent enzyme fold. It consisted of two distinct domains, both of which show the typical open twisted α/β structure. A PLP cofactor was located in the crevice between the two domains, which was attached to Lys52 by a Schiff-base linkage. Unique residue substitutions (Gly78, Pro79, Ser146, Ser147 and Thr312) were discovered at the catalytic site of RmSDH by comparison of structures of RmSDH and other reported eukaryotic L-serine ammonia-lyases. Optimal pH and temperature of the purified RmSDH were 7.5 and 40 °C, respectively. It was stable in the pH range of 7.0–9.0 and at temperatures below 40 °C. This is the first crystal structure of a fungal L-serine ammonia-lyase. It will be useful to study the catalytic mechanism of β-elimination enzymes and will provide a basis for further enzyme engineering. - Highlights: • The crystal structure of a fungal L-serine ammonia-lyase (RmSDH) was solved. • Five unique residue substitutions are found at the catalytic site of RmSDH. • RmSDH was expressed in Pichia. pastoris and biochemically characterized. • RmSDH has potential application in splitting D/L-serine.

  9. UVA, UVB and UVC Light Enhances the Biosynthesis of Phenolic Antioxidants in Fresh-Cut Carrot through a Synergistic Effect with Wounding

    Directory of Open Access Journals (Sweden)

    Bernadeth B. Surjadinata

    2017-04-01

    Full Text Available Previously, we found that phenolic content and antioxidant capacity (AOX in carrots increased with wounding intensity. It was also reported that UV radiation may trigger the phenylpropanoid metabolism in plant tissues. Here, we determined the combined effect of wounding intensity and UV radiation on phenolic compounds, AOX, and the phenylalanine ammonia-lyase (PAL activity of carrots. Accordingly, phenolic content, AOX, and PAL activity increased in cut carrots with the duration of UVC radiation, whereas whole carrots showed no increase. Carrot pies showed a higher increase compared to slices and shreds. Phenolics, AOX, and PAL activity also increased in cut carrots exposed to UVA or UVB. The major phenolics were chlorogenic acid and its isomers, ferulic acid, and isocoumarin. The type of UV radiation affected phenolic profiles. Chlorogenic acid was induced by all UV radiations but mostly by UVB and UVC, ferulic acid was induced by all UV lights to comparable levels, while isocoumarin and 4,5-diCQA was induced mainly by UVB and UVC compared to UVA. In general, total phenolics correlated linearly with AOX for all treatments. A reactive oxygen species (ROS mediated hypothetical mechanism explaining the synergistic effect of wounding and different UV radiation stresses on phenolics accumulation in plants is herein proposed.

  10. Cystathionine .gamma.-lyase: Clinical, metabolic, genetic, and structural studies

    Czech Academy of Sciences Publication Activity Database

    Kraus, J. P.; Hašek, Jindřich; Kožich, V.; Collard, R.; Venezia, S.; Janošíková, B.; Wang, J.; Stabler, S. P.; Allen, R. H.; Jakobs, C.; Finn, C. T.; Chien, Y. H.; Hwu, W. L.; Hegele, R. A.; Mudd, S. H.

    2009-01-01

    Roč. 97, č. 4 (2009), s. 250-259 ISSN 1096-7192 R&D Projects: GA ČR GA305/07/1073 Institutional research plan: CEZ:AV0Z40500505 Keywords : cystathionine gamma-lyase * cystathioninuria * hypercystathioninemia Subject RIV: CD - Macromolecular Chemistry Impact factor: 2.897, year: 2009

  11. Molecular cloning and characterization of l-methionine γ-lyase from Streptomyces avermitilis.

    Science.gov (United States)

    Kudou, Daizou; Yasuda, Eri; Hirai, Yoshiyuki; Tamura, Takashi; Inagaki, Kenji

    2015-10-01

    A pyridoxal 5'-phosphate-dependent methionine γ-lyase (MGL) was cloned from Streptomyces avermitilis catalyzed the degradation of methionine to α-ketobutyrate, methanethiol, and ammonia. The sav7062 gene (1,242 bp) was corresponded to 413 amino acid residues with a molecular mass of 42,994 Da. The deduced amino acid sequence showed a high degree of similarity to those of other MGL enzymes. The sav7062 gene was overexpressed in Escherichia coli. The enzyme was purified to homogeneity and exhibited the MGL catalytic activities. We cloned the enzyme that has the MGL activity in Streptomyces for the first time. Copyright © 2015 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  12. Sphingosine-1-Phosphate Lyase Deficient Cells as a Tool to Study Protein Lipid Interactions.

    Directory of Open Access Journals (Sweden)

    Mathias J Gerl

    Full Text Available Cell membranes contain hundreds to thousands of individual lipid species that are of structural importance but also specifically interact with proteins. Due to their highly controlled synthesis and role in signaling events sphingolipids are an intensely studied class of lipids. In order to investigate their metabolism and to study proteins interacting with sphingolipids, metabolic labeling based on photoactivatable sphingoid bases is the most straightforward approach. In order to monitor protein-lipid-crosslink products, sphingosine derivatives containing a reporter moiety, such as a radiolabel or a clickable group, are used. In normal cells, degradation of sphingoid bases via action of the checkpoint enzyme sphingosine-1-phosphate lyase occurs at position C2-C3 of the sphingoid base and channels the resulting hexadecenal into the glycerolipid biosynthesis pathway. In case the functionalized sphingosine looses the reporter moiety during its degradation, specificity towards sphingolipid labeling is maintained. In case degradation of a sphingosine derivative does not remove either the photoactivatable or reporter group from the resulting hexadecenal, specificity towards sphingolipid labeling can be achieved by blocking sphingosine-1-phosphate lyase activity and thus preventing sphingosine derivatives to be channeled into the sphingolipid-to-glycerolipid metabolic pathway. Here we report an approach using clustered, regularly interspaced, short palindromic repeats (CRISPR-associated nuclease Cas9 to create a sphingosine-1-phosphate lyase (SGPL1 HeLa knockout cell line to disrupt the sphingolipid-to-glycerolipid metabolic pathway. We found that the lipid and protein compositions as well as sphingolipid metabolism of SGPL1 knock-out HeLa cells only show little adaptations, which validates these cells as model systems to study transient protein-sphingolipid interactions.

  13. Study design and protocol for a mixed methods evaluation of an intervention to reduce and break up sitting time in primary school classrooms in the UK: The CLASS PAL (Physically Active Learning) Programme

    OpenAIRE

    Routen, Ash C; Biddle, Stuart J H; Bodicoat, Danielle H; Cale, Lorraine; Clemes, Stacy; Edwardson, Charlotte L; Glazebrook, Cris; Harrington, Deirdre M; Khunti, Kamlesh; Pearson, Natalie; Salmon, Jo; Sherar, Lauren B

    2017-01-01

    Introduction Children engage in a high volume of sitting in school, particularly in the classroom. A number of strategies, such as physically active lessons (termed movement integration (MI)), have been developed to integrate physical activity into this learning environment; however, no single approach is likely to meet the needs of all pupils and teachers. This protocol outlines an implementation study of a primary school-based MI intervention: CLASS PAL (Physically Active Learning) programm...

  14. Utilization of Glyphosate as Phosphate Source: Biochemistry and Genetics of Bacterial Carbon-Phosphorus Lyase

    Science.gov (United States)

    Zechel, David L.; Jochimsen, Bjarne

    2014-01-01

    SUMMARY After several decades of use of glyphosate, the active ingredient in weed killers such as Roundup, in fields, forests, and gardens, the biochemical pathway of transformation of glyphosate phosphorus to a useful phosphorus source for microorganisms has been disclosed. Glyphosate is a member of a large group of chemicals, phosphonic acids or phosphonates, which are characterized by a carbon-phosphorus bond. This is in contrast to the general phosphorus compounds utilized and metabolized by microorganisms. Here phosphorus is found as phosphoric acid or phosphate ion, phosphoric acid esters, or phosphoric acid anhydrides. The latter compounds contain phosphorus that is bound only to oxygen. Hydrolytic, oxidative, and radical-based mechanisms for carbon-phosphorus bond cleavage have been described. This review deals with the radical-based mechanism employed by the carbon-phosphorus lyase of the carbon-phosphorus lyase pathway, which involves reactions for activation of phosphonate, carbon-phosphorus bond cleavage, and further chemical transformation before a useful phosphate ion is generated in a series of seven or eight enzyme-catalyzed reactions. The phn genes, encoding the enzymes for this pathway, are widespread among bacterial species. The processes are described with emphasis on glyphosate as a substrate. Additionally, the catabolism of glyphosate is intimately connected with that of aminomethylphosphonate, which is also treated in this review. Results of physiological and genetic analyses are combined with those of bioinformatics analyses. PMID:24600043

  15. Enantioselective Synthesis of Various Cyanohydrins Using Covalently Immobilized Preparations of Hydroxynitrile Lyase from Prunus dulcis.

    Science.gov (United States)

    Alagöz, Dilek; Tükel, S Seyhan; Yildirim, Deniz

    2015-11-01

    The carrier-based and carrier-free (cross-linked enzyme aggregate) covalent immobilizations of Prunus dulcis hydroxynitrile lyase were investigated. The immobilized preparations were tested for enantioselective carbon-carbon bond formation activity in the biphasic medium. Of the tested preparations, only cross-linked enzyme aggregate of P. dulcis hydroxynitrile lyase (PdHNL-CLEA) achieved the synthesis of (R)-mandelonitrile with 93% yield and 99% enantiopurity. PdHNL-CLEA was also used in the synthesis of various (R)-cyanohydrins from corresponding aldehydes/ketones and hydrocyanic acid. When 4-methoxybenzaldehyde, 4-methyl benzaldehyde, and 4-hydroxybenzaldehyde were used as substrates, the yield-enantiomeric excess of corresponding (R)-cyanohydrins were obtained as 95-95, 85-79, and 2-25%, respectively, after 96 h at pH 4.0 and 5 °C. For acetophenone, 4-fluoroacetophenone, 4-chloroacetophenone, 4-bromoacetophenone, and 4-iodoacetophenone, the yield-enantiomeric excess of corresponding (R)-cyanohydrins were 1-99, 20-84, 11-95, 5-99, and 3-24%, respectively at the same conditions. The results demonstrate PdHNL-CLEA can be effectively used in the synthesis of (R)-mandelonitrile.

  16. Uncoupled defense gene expression and antimicrobial alkaloid accumulation in elicited opium poppy cell cultures.

    Science.gov (United States)

    Facchini, P J; Johnson, A G; Poupart, J; de Luca, V

    1996-01-01

    Treatment of opium poppy (Papaver somniferum L.) cell cultures with autoclaved mycelial homogenates of Botrytis sp. resulted in the accumulation of sanguinarine. Elicitor treatment also caused a rapid and transient induction in the activity of tyrosine/dopa decarboxylase (TYDC, EC 4.1.1.25), which catalyzes the conversion of L-tyrosine and L-dopa to tyramine and dopamine, respectively, the first steps in sanguinarine biosynthesis. TYDC genes were differentially expressed in response to elicitor treatment. TYDC1-like mRNA levels were induced rapidly but declined to near baseline levels within 5 h. In contrast, TYDC2-like transcript levels increased more slowly but were sustained for an extended period. Induction of TYDC mRNAs preceded that of phenylalanine ammonia-lyase (PAL, EC 4.3.1.5) mRNAs. An elicitor preparation from Pythium aphanidermatum was less effective in the induction of TYDC mRNA levels and alkaloid accumulation; however, both elicitors equally induced accumulation of PAL transcripts. In contrast, treatment with methyl jasmonate resulted in an induction of TYDC but not PAL mRNAs. The calmodulin antagonist N-(6-aminohexyl)-5-chloro-1-naphthalenesulfonamide and the protein kinase inhibitor staurosporine partially blocked the fungal elicitor-induced accumulation of sanguinarine. However, only staurosporine and okadaic acid, an inhibitor of protein phosphatases 1 and 2A, blocked the induction of TYDC1-like transcript levels, but they did not block the induction of TYDC2-like or PAL transcript levels. These data suggest that activation mechanisms for PAL, TYDC, and some later sanguinarine biosynthetic enzymes are uncoupled. PMID:8754678

  17. Content Analysis Schedule for Bilingual Education Programs: Proyecto PAL.

    Science.gov (United States)

    Gonzalez, Castor

    This content analysis schedule for "Proyecto PAL" in San Jose, California, presents information on the history, funding, and scope of the project. Included are sociolinguistic process variables such as the native and dominant languages of students and their interaction. Information is provided on staff selection and the linguistic…

  18. Priming ammonia lyases and aminomutases for industrial and therapeutic applications

    NARCIS (Netherlands)

    Heberling, Matthew M.; Wu, Bian; Bartsch, Sebastian; Janssen, Dick B.

    Ammonia lyases (AL) and aminomutases (AM) are emerging in green synthetic routes to chiral amines and an AL is being explored as an enzyme therapeutic for treating phenylketonuria and cancer. Although the restricted substrate range of the wild-type enzymes limits their widespread application, the

  19. Vascular expression of endothelial antigen PAL-E indicates absence of blood-ocular barriers in the normal eye

    NARCIS (Netherlands)

    Schlingemann, R. O.; Hofman, P.; Anderson, L.; Troost, D.; van der Gaag, R.

    1997-01-01

    The endothelium-specific antigen PAL-E is expressed in capillaries and veins throughout the body with the exception of the brain, where the antigen is absent from anatomical sites with a patent blood-brain barrier. In this study we determined vascular endothelial staining for PAL-E in the normal eye

  20. Correlation between the cystathionine-r-lyase (CES) and the ...

    African Journals Online (AJOL)

    Background: The infection of Helicobacter pylori (H. pylori) is one of the most important causes of gastric ulcer disease. The role of hydrogen sulfide (H2S) production in H. pylori-induced gastric ulcer disease. Aim: The expression of cystathionine-γ-lyase (CSE) was determined, and correlated with the severity of gastric ulcer ...

  1. PAL driven organizational learning theory and practices a light on learning journey of organizations

    CERN Document Server

    Chuah, Kong

    2015-01-01

    Presenting an innovative concept and approach for organization management, this book serves to document an organization’s journey towards the ultimate goal of learning organization. This book also shares the experience on how a OL framework built on established learning theories, could be used effectively, overcoming many of the barriers in a real industrial setting. Utilizing a ready-to-use tool called Project Action Learning (PAL) to analyze real life case studies, the authors introduce a framework that allows teams of people to work and learn over the course of business projects. Equal emphasis is placed on the achievement of pre-set project outcomes and the learning objectives of the participants. In addition, a long term organizational learning strategy is put forward and the necessary supporting infrastructure, in the form of four ‘PAL Pillars’, is described. The concepts and development of the PAL driven Organizational Learning model are inspired by, and grounded in, Western and Eastern business ...

  2. Sphingosine 1-phosphate lyase enzyme assay using a BODIPY-labeled substrate

    International Nuclear Information System (INIS)

    Bandhuvula, Padmavathi; Li Zaiguo; Bittman, Robert; Saba, Julie D.

    2009-01-01

    Sphingosine 1-phosphate lyase (SPL) is responsible for the irreversible catabolism of sphingosine 1-phosphate, which signals through five membrane receptors to mediate cell stress responses, angiogenesis, and lymphocyte trafficking. The standard assay for SPL activity utilizes a radioactive dihydrosphingosine 1-phosphate substrate and is expensive and cumbersome. In this study, we describe an SPL assay that employs an ω-labeled BODIPY-sphingosine 1-phosphate substrate, allowing fluorescent product detection by HPLC and incorporating advantages of the BODIPY fluorophore. The major aldehyde product is confirmed by reaction with 2,4-dinitrophenylhydrazine. The SPL-catalyzed reaction is linear over a 30 min time period and yields a K m of 35 μM for BODIPY-sphingosine 1-phosphate.

  3. Status of PPI (Pohang Photo-Injector) for PAL XFEL

    CERN Document Server

    Park, Sung-Ju; Oh, Jong-Seok; Park, Chong-Do; Park Jang Ho; Soo Ko In; Wang, Xijie; Woon Parc, Yong

    2005-01-01

    A X-Ray Free Electron Laser (XFEL) project based on the Self-Amplified Spontaneous Emission (SASE) is under progress at the Pohang Accelerator Laboratory (PAL). One of the critical R&D for the PAL XFEL* is to develop the Pohang Photo-Injector (PPI) which is required to deliver electron beams with normalized emittance < 1.5 mm-mrad. In order to achieve the required beam quality with high stability and reliability, we will use photocathode with quantum efficiency > 0.1 % and long lifetime. This will greatly lessen the laser energy requirement for producing flat-top UV pulses, and open the possibility of using only regenerative amplifiers (RGAs) to drive the photocathode RF gun. The RGAs can produce mJs output with much better stability than multi-pass amplifiers. Both the Cs2Te and Mg are under consideration for the possible photo-cathode. To demonstrate the suitability of the Mg and Cs2Te for the future 4th generation light source application, an improved BNL-type S-band RF gun with a high-pe...

  4. Structural Insights Into The Bacterial Carbon-Phosphorus Lyase Machinery

    DEFF Research Database (Denmark)

    Brodersen, Ditlev Egeskov

    the proteins encoded in the phn operon act in concert to catabolise phosphonate remain unknown. We have determined the crystal structure of a 240 kDa Escherichia coli carbon-phosphorus lyase core complex at 1.7 Å and show that it comprises a highly intertwined network of subunits with several unexpected......Phosphonate compounds act as a nutrient source for some microorganisms when phosphate is limiting but require a specialised enzymatic machinery due to the presence of the highly stable carbon-phosphorus bond. Despite the fundamental importance to microbial metabolism, the details of how...... structural features. The complex contains at least two different active sites and suggest a revision of current models of carbon-phosphorus bond cleavage. Using electron microscopy, we map the binding site of an additional protein subunit, which may use ATP for driving conformational changes during...

  5. Quality of Life in Patients With Brain Metastases Using the EORTC QLQ-BN20+2 and QLQ-C15-PAL

    International Nuclear Information System (INIS)

    Caissie, Amanda; Nguyen, Janet; Chen, Emily; Zhang Liying; Sahgal, Arjun; Clemons, Mark; Kerba, Marc; Arnalot, Palmira Foro; Danjoux, Cyril; Tsao, May; Barnes, Elizabeth; Holden, Lori; Danielson, Brita; Chow, Edward

    2012-01-01

    Purpose: The 20-item European Organisation for Research and Treatment of Cancer Quality of Life Questionnaire–Brain Neoplasm (QLQ-BN20) is a validated quality-of-life (QOL) questionnaire for patients with primary brain tumors. The European Organisation for Research and Treatment of Cancer Quality of Life Questionnaire–Core 15 Palliative (QLQ-C15-PAL) core palliative questionnaire is a 15-item version of the core 30-item QLQ-C30 and was developed to decrease the burden on patients with advanced cancer. The combination of the QLQ-BN20 and QLQ-C30 to assess QOL may be too burdensome for patients. The primary aim of this study was to assess QOL in patients before and after treatment for brain metastases using the QLQ-BN20+2 and QLQ-C15-PAL, a version of the QLQ-BN20 questionnaire with 2 additional questions assessing cognitive functioning that were not addressed in the QLQ-C15-PAL. Methods and Materials: Patients with brain metastases completed the QLQ-C15-PAL and QLQ-BN20+2 questionnaires to assess QOL before and 1 month after radiation. Linear regression analysis was used to assess changes in QOL scores over time, as well as to explore associations between the QLQ-BN20+2 and QLQ-C15-PAL scales, patient demographics, and clinical variables. Spearman correlation assessed associations between the QLQ-BN20+2 and QLQ-C15-PAL scales. Results: Among 108 patients, the majority (55%) received whole-brain radiotherapy only, with 65% of patients completing follow-up at 1 month after treatment. The most prominent symptoms at baseline were future uncertainty (QLQ-BN20+2) and fatigue (QLQ-C15-PAL). After treatment, significant improvement was seen for the QLQ-C15-PAL insomnia scale, as well as the QLQ-BN20+2 scales of future uncertainty, visual disorder, and concentration difficulty. Baseline Karnofsky Performance Status was negatively correlated to QLQ-BN20+2 motor dysfunction but positively related to QLQ-C15-PAL physical functioning and QLQ-BN20+2 cognitive functioning at

  6. Quality of Life in Patients With Brain Metastases Using the EORTC QLQ-BN20+2 and QLQ-C15-PAL

    Energy Technology Data Exchange (ETDEWEB)

    Caissie, Amanda; Nguyen, Janet; Chen, Emily; Zhang Liying [Rapid Response Radiotherapy Program, Odette Cancer Centre, Sunnybrook Health Sciences Centre, University of Toronto, Toronto, Ontario (Canada); Sahgal, Arjun [Rapid Response Radiotherapy Program, Odette Cancer Centre, Sunnybrook Health Sciences Centre, University of Toronto, Toronto, Ontario (Canada); Department of Radiation Oncology, Princess Margaret Hospital, University of Toronto, Toronto, Ontario (Canada); Clemons, Mark [Department of Medical Oncology, Ottawa Hospital Cancer Centre, Ottawa, Ontario (Canada); Kerba, Marc [Department of Radiation Oncology, Tom Baker Cancer Centre, Calgary, Alberta (Canada); Arnalot, Palmira Foro [Parc de Salut Mar Hospital de l' Esperanca, Barcelona (Spain); Danjoux, Cyril; Tsao, May; Barnes, Elizabeth; Holden, Lori [Rapid Response Radiotherapy Program, Odette Cancer Centre, Sunnybrook Health Sciences Centre, University of Toronto, Toronto, Ontario (Canada); Danielson, Brita [Department of Radiation Oncology, Cross Cancer Institute, Edmonton, Alberta (Canada); Chow, Edward, E-mail: edward.chow@sunnybrook.ca [Rapid Response Radiotherapy Program, Odette Cancer Centre, Sunnybrook Health Sciences Centre, University of Toronto, Toronto, Ontario (Canada)

    2012-07-15

    Purpose: The 20-item European Organisation for Research and Treatment of Cancer Quality of Life Questionnaire-Brain Neoplasm (QLQ-BN20) is a validated quality-of-life (QOL) questionnaire for patients with primary brain tumors. The European Organisation for Research and Treatment of Cancer Quality of Life Questionnaire-Core 15 Palliative (QLQ-C15-PAL) core palliative questionnaire is a 15-item version of the core 30-item QLQ-C30 and was developed to decrease the burden on patients with advanced cancer. The combination of the QLQ-BN20 and QLQ-C30 to assess QOL may be too burdensome for patients. The primary aim of this study was to assess QOL in patients before and after treatment for brain metastases using the QLQ-BN20+2 and QLQ-C15-PAL, a version of the QLQ-BN20 questionnaire with 2 additional questions assessing cognitive functioning that were not addressed in the QLQ-C15-PAL. Methods and Materials: Patients with brain metastases completed the QLQ-C15-PAL and QLQ-BN20+2 questionnaires to assess QOL before and 1 month after radiation. Linear regression analysis was used to assess changes in QOL scores over time, as well as to explore associations between the QLQ-BN20+2 and QLQ-C15-PAL scales, patient demographics, and clinical variables. Spearman correlation assessed associations between the QLQ-BN20+2 and QLQ-C15-PAL scales. Results: Among 108 patients, the majority (55%) received whole-brain radiotherapy only, with 65% of patients completing follow-up at 1 month after treatment. The most prominent symptoms at baseline were future uncertainty (QLQ-BN20+2) and fatigue (QLQ-C15-PAL). After treatment, significant improvement was seen for the QLQ-C15-PAL insomnia scale, as well as the QLQ-BN20+2 scales of future uncertainty, visual disorder, and concentration difficulty. Baseline Karnofsky Performance Status was negatively correlated to QLQ-BN20+2 motor dysfunction but positively related to QLQ-C15-PAL physical functioning and QLQ-BN20+2 cognitive functioning at

  7. Diversity of function in the isocitrate lyase enzyme superfamily: the Dianthus caryophyllus petal death protein cleaves alpha-keto and alpha-hydroxycarboxylic acids.

    Science.gov (United States)

    Lu, Zhibing; Feng, Xiaohua; Song, Ling; Han, Ying; Kim, Alexander; Herzberg, Osnat; Woodson, William R; Martin, Brian M; Mariano, Patrick S; Dunaway-Mariano, Debra

    2005-12-20

    The work described in this paper was carried out to define the chemical function a new member of the isocitrate lyase enzyme family derived from the flowering plant Dianthus caryophyllus. This protein (Swiss-Prot entry Q05957) is synthesized in the senescent flower petals and is named the "petal death protein" or "PDP". On the basis of an analysis of the structural contexts of sequence markers common to the C-C bond lyases of the isocitrate lyase/phosphoenolpyruvate mutase superfamily, a substrate screen that employed a (2R)-malate core structure was designed. Accordingly, stereochemically defined C(2)- and C(3)-substituted malates were synthesized and tested as substrates for PDP-catalyzed cleavage of the C(2)-C(3) bond. The screen identified (2R)-ethyl, (3S)-methylmalate, and oxaloacetate [likely to bind as the hydrate, C(2)(OH)(2) gem-diol] as the most active substrates (for each, k(cat)/K(m) = 2 x 10(4) M(-)(1) s(-)(1)). In contrast to the stringent substrate specificities previously observed for the Escherichia coli isocitrate and 2-methylisocitrate lyases, the PDP tolerated hydrogen, methyl, and to a much lesser extent acetate substituents at the C(3) position (S configuration only) and hydoxyl, methyl, ethyl, propyl, and to a much lesser extent isobutyl substituents at C(2) (R configuration only). It is hypothesized that PDP functions in oxalate production in Ca(2+) sequestering and/or in carbon scavenging from alpha-hydroxycarboxylate catabolites during the biochemical transition accompanying petal senescence.

  8. Characterization of the N-linked glycosylation site of recombinant pectate lyase

    NARCIS (Netherlands)

    Colangelo, J.; Licon, V.; Benen, J.A.E.; Visser, J.; Bergmann, C.; Orlando, R.

    1999-01-01

    Recombinant pectate lyase from Aspergillus niger was overexpressed in Aspergillus nidulans. The two recombinant proteins produced differed in molecular mass by 1200 Da, which suggested that the larger molecular weight protein was glycosylated. The deduced amino acid sequence was searched for

  9. Structures of the human Pals1 PDZ domain with and without ligand suggest gated access of Crb to the PDZ peptide-binding groove

    Energy Technology Data Exchange (ETDEWEB)

    Ivanova, Marina E.; Fletcher, Georgina C.; O’Reilly, Nicola; Purkiss, Andrew G.; Thompson, Barry J. [Cancer Research UK, 44 Lincoln’s Inn Fields, London WC2A 3LY (United Kingdom); McDonald, Neil Q., E-mail: neil.mcdonald@cancer.org.uk [Cancer Research UK, 44 Lincoln’s Inn Fields, London WC2A 3LY (United Kingdom); Birkbeck College, University of London, Malet Street, London WC1E 7HX (United Kingdom)

    2015-03-01

    This study characterizes the interaction between the carboxy-terminal (ERLI) motif of the essential polarity protein Crb and the Pals1/Stardust PDZ-domain protein. Structures of human Pals1 PDZ with and without a Crb peptide are described, explaining the highly conserved nature of the ERLI motif and revealing a sterically blocked peptide-binding groove in the absence of ligand. Many components of epithelial polarity protein complexes possess PDZ domains that are required for protein interaction and recruitment to the apical plasma membrane. Apical localization of the Crumbs (Crb) transmembrane protein requires a PDZ-mediated interaction with Pals1 (protein-associated with Lin7, Stardust, MPP5), a member of the p55 family of membrane-associated guanylate kinases (MAGUKs). This study describes the molecular interaction between the Crb carboxy-terminal motif (ERLI), which is required for Drosophila cell polarity, and the Pals1 PDZ domain using crystallography and fluorescence polarization. Only the last four Crb residues contribute to Pals1 PDZ-domain binding affinity, with specificity contributed by conserved charged interactions. Comparison of the Crb-bound Pals1 PDZ structure with an apo Pals1 structure reveals a key Phe side chain that gates access to the PDZ peptide-binding groove. Removal of this side chain enhances the binding affinity by more than fivefold, suggesting that access of Crb to Pals1 may be regulated by intradomain contacts or by protein–protein interaction.

  10. An application of programmatic assessment for learning (PAL) system for general practice training.

    Science.gov (United States)

    Schuwirth, Lambert; Valentine, Nyoli; Dilena, Paul

    2017-01-01

    Aim: Programmatic assessment for learning (PAL) is becoming more and more popular as a concept but its implementation is not without problems. In this paper we describe the design principles behind a PAL program in a general practice training context. Design principles: The PAL program was designed to optimise the meaningfulness of assessment information for the registrar and to make him/her use that information to self regulate their learning. The main principles in the program were cognitivist and transformative. The main cognitive principles we used were fostering the understanding of deep structures and stimulating transfer by making registrars constantly connect practice experiences with background knowledge. Ericsson's deliberate practice approach was built in with regard to the provision of feedback combined with Pintrich's model of self regulation. Mezirow's transformative learning and insights from social network theory on collaborative learning were used to support the registrars in their development to become GP professionals. Finally the principal of test enhanced learning was optimised. Epilogue: We have provided this example explain the design decisions behind our program, but not want to present our program as the solution to any given situation.

  11. Subcellular Targeting of Methylmercury Lyase Enhances Its Specific Activity for Organic Mercury Detoxification in Plants1

    Science.gov (United States)

    Bizily, Scott P.; Kim, Tehryung; Kandasamy, Muthugapatti K.; Meagher, Richard B.

    2003-01-01

    Methylmercury is an environmental pollutant that biomagnifies in the aquatic food chain with severe consequences for humans and other animals. In an effort to remove this toxin in situ, we have been engineering plants that express the bacterial mercury resistance enzymes organomercurial lyase MerB and mercuric ion reductase MerA. In vivo kinetics experiments suggest that the diffusion of hydrophobic organic mercury to MerB limits the rate of the coupled reaction with MerA (Bizily et al., 2000). To optimize reaction kinetics for organic mercury compounds, the merB gene was engineered to target MerB for accumulation in the endoplasmic reticulum and for secretion to the cell wall. Plants expressing the targeted MerB proteins and cytoplasmic MerA are highly resistant to organic mercury and degrade organic mercury at 10 to 70 times higher specific activity than plants with the cytoplasmically distributed wild-type MerB enzyme. MerB protein in endoplasmic reticulum-targeted plants appears to accumulate in large vesicular structures that can be visualized in immunolabeled plant cells. These results suggest that the toxic effects of organic mercury are focused in microenvironments of the secretory pathway, that these hydrophobic compartments provide more favorable reaction conditions for MerB activity, and that moderate increases in targeted MerB expression will lead to significant gains in detoxification. In summary, to maximize phytoremediation efficiency of hydrophobic pollutants in plants, it may be beneficial to target enzymes to specific subcellular environments. PMID:12586871

  12. Inducers of resistance and silicon on the activity of defense enzymes in the soybean-Phakopsora pachyrhizi interaction

    Directory of Open Access Journals (Sweden)

    Maria Fernanda Antunes da Cruz

    2013-06-01

    Full Text Available This study aimed to determine the effect of jasmonic acid (JA, Acibenzolar-S-Methyl (ASM and calcium silicate (a source of soluble silicon, Si, on the potentiation of soybean resistance to Asian soybean rust (ASR. The ASR severity was significantly reduced on plants sprayed with ASM or supplied with Si in comparison to plants sprayed with JA or deionized water. For chitinases (CHI, significant differences in activity between non-inoculated and inoculated plants sprayed with deionized water or with ASM occurred at 72 hours after inoculation (hai, at 24 and 72 hai when sprayed with JA and at 141 hai when supplied with Si. For β-1,3-glucanases (GLU, significant differences in activity between non-inoculated and inoculated plants sprayed with deionized water occurred at 24, 48 and 141 hai, but not until 72 for plants sprayed with ASM. For phenylalanine ammonia-lyases (PAL, significant differences in activity between non-inoculated and inoculated plants occurred only for plants sprayed with ASM at 72 and 141 hai. In conclusion, the ASR symptoms can be mild on plants sprayed with ASM or supplied with Si and that this amelioration likely involved the defense enzymes.

  13. Probing the structure of glucan lyases – the lytic members of GH31 - by sequence analysis, circular dichroism and proteolysis

    DEFF Research Database (Denmark)

    Ernst, Heidi; Lo Leggio, Leila; Yu, Shukun

    2005-01-01

    Glucan lyase (GL) is a polysaccharide lyase with unique characteristics. It is involved in an alternative pathway for the degradation of alpha-glucans, the anhydrofructose pathway. Sequence similarity suggests that this lytic enzyme belongs to glycoside hydrolase family 31, for which until very r...

  14. Epigenetic regulation of pro-inflammatory cytokine secretion by sphingosine 1-phosphate (S1P) in acute lung injury: Role of S1P lyase.

    Science.gov (United States)

    Ebenezer, David L; Fu, Panfeng; Suryadevara, Vidyani; Zhao, Yutong; Natarajan, Viswanathan

    2017-01-01

    Cellular level of sphingosine-1-phosphate (S1P), the simplest bioactive sphingolipid, is tightly regulated by its synthesis catalyzed by sphingosine kinases (SphKs) 1 & 2 and degradation mediated by S1P phosphatases, lipid phosphate phosphatases, and S1P lyase. The pleotropic actions of S1P are attributed to its unique inside-out (extracellular) signaling via G-protein-coupled S1P1-5 receptors, and intracellular receptor independent signaling. Additionally, S1P generated in the nucleus by nuclear SphK2 modulates HDAC1/2 activity, regulates histone acetylation, and transcription of pro-inflammatory genes. Here, we present data on the role of S1P lyase mediated S1P signaling in regulating LPS-induced inflammation in lung endothelium. Blocking S1P lyase expression or activity attenuated LPS-induced histone acetylation and secretion of pro-inflammatory cytokines. Degradation of S1P by S1P lyase generates Δ2-hexadecenal and ethanolamine phosphate and the long-chain fatty aldehyde produced in the cytoplasmic compartment of the endothelial cell seems to modulate histone acetylation pattern, which is different from the nuclear SphK2/S1P signaling and inhibition of HDAC1/2. These in vitro studies suggest that S1P derived long-chain fatty aldehyde may be an epigenetic regulator of pro-inflammatory genes in sepsis-induced lung inflammation. Trapping fatty aldehydes and other short chain aldehydes such as 4-hydroxynonenal derived from S1P degradation and lipid peroxidation, respectively by cell permeable agents such as phloretin or other aldehyde trapping agents may be useful in treating sepsis-induced lung inflammation via modulation of histone acetylation. . Copyright © 2016 Elsevier Ltd. All rights reserved.

  15. Lactic acid bacteria involved in cocoa beans fermentation from Ivory Coast: Species diversity and citrate lyase production.

    Science.gov (United States)

    Ouattara, Hadja D; Ouattara, Honoré G; Droux, Michel; Reverchon, Sylvie; Nasser, William; Niamke, Sébastien L

    2017-09-01

    Microbial fermentation is an indispensable process for high quality chocolate from cocoa bean raw material. lactic acid bacteria (LAB) are among the major microorganisms responsible for cocoa fermentation but their exact role remains to be elucidated. In this study, we analyzed the diversity of LAB in six cocoa producing regions of Ivory Coast. Ribosomal 16S gene sequence analysis showed that Lactobacillus plantarum and Leuconostoc mesenteroides are the dominant LAB species in these six regions. In addition, other species were identified as the minor microbial population, namely Lactobacillus curieae, Enterococcus faecium, Fructobacillus pseudoficulneus, Lactobacillus casei, Weissella paramesenteroides and Weissella cibaria. However, in each region, the LAB microbial population was composed of a restricted number of species (maximum 5 species), which varied between the different regions. LAB implication in the breakdown of citric acid was investigated as a fundamental property for a successful cocoa fermentation process. High citrate lyase producer strains were characterized by rapid citric acid consumption, as revealed by a 4-fold decrease in citric acid concentration in the growth medium within 12h, concomitant with an increase in acetic acid and lactic acid concentration. The production of citrate lyase was strongly dependent on environmental conditions, with optimum production at acidic pH (pHfermentation. This study reveals that one of the major roles of LAB in the cocoa fermentation process involves the breakdown of citric acid during the early stage of cocoa fermentation through the activity of citrate lyase. Copyright © 2017 Elsevier B.V. All rights reserved.

  16. The Phenylpropanoid Pathway and Lignin in Defense against Ganoderma boninense Colonized Root Tissues in Oil Palm (Elaeis guineensis Jacq.).

    Science.gov (United States)

    Govender, Nisha T; Mahmood, Maziah; Seman, Idris A; Wong, Mui-Yun

    2017-01-01

    Basal stem rot, caused by the basidiomycete fungus, Ganoderma boninense , is an economically devastating disease in Malaysia. Our study investigated the changes in lignin content and composition along with activity and expression of the phenylpropanoid pathway enzymes and genes in oil palm root tissues during G. boninense infection. We sampled control (non-inoculated) and infected (inoculated) seedlings at seven time points [1, 2, 3, 4, 8, and 12 weeks post-inoculation (wpi)] in a randomized design. The expression profiles of phenylalanine ammonia lyase (PAL), cinnamyl alcohol dehydrogenase (CAD), and peroxidase (POD) genes were monitored at 1, 2, and 3 wpi using real-time quantitative polymerase chain reaction. Seedlings at 4, 8, and 12 wpi were screened for lignin content, lignin composition, enzyme activities (PAL, CAD, and POD), growth (weight and height), and disease severity (DS). Gene expression analysis demonstrated up-regulation of PAL, CAD, and POD genes in the infected seedlings, relative to the control seedlings at 1, 2, and 3 wpi. At 2 and 3 wpi, CAD showed highest transcript levels compared to PAL and POD. DS increased progressively throughout sampling, with 5, 34, and 69% at 4, 8, and 12 wpi, respectively. Fresh weight and height of the infected seedlings were significantly lower compared to the control seedlings at 8 and 12 wpi. Lignin content of the infected seedlings at 4 wpi was significantly higher than the control seedlings, remained elicited with no change at 8 wpi, and then collapsed with a significant reduction at 12 wpi. The nitrobenzene oxidation products of oil palm root lignin yielded both syringyl and guaiacyl monomers. Accumulation of lignin in the infected seedlings was in parallel to increased syringyl monomers, at 4 and 8 wpi. The activities of PAL and CAD enzymes in the infected seedlings at DS = 5-34% were significantly higher than the control seedlings and thereafter collapsed at DS = 69%.

  17. Microbial β-etherases and glutathione lyases for lignin valorisation in biorefineries: current state and future perspectives.

    Science.gov (United States)

    Husarcíková, Jana; Voß, Hauke; Domínguez de María, Pablo; Schallmey, Anett

    2018-05-04

    Lignin is the major aromatic biopolymer in nature, and it is considered a valuable feedstock for the future supply of aromatics. Hence, its valorisation in biorefineries is of high importance, and various chemical and enzymatic approaches for lignin depolymerisation have been reported. Among the enzymes known to act on lignin, β-etherases offer the possibility for a selective cleavage of the β-O-4 aryl ether bonds present in lignin. These enzymes, together with glutathione lyases, catalyse a reductive, glutathione-dependent ether bond cleavage displaying high stereospecificity. β-Etherases and glutathione lyases both belong to the superfamily of glutathione transferases, and several structures have been solved recently. Additionally, different approaches for their application in lignin valorisation have been reported in the last years. This review gives an overview on the current knowledge on β-etherases and glutathione lyases, their biochemical and structural features, and critically discusses their potential for application in biorefineries.

  18. Charged defects in chalcogenide vitreous semiconductors studied with combined Raman scattering and PALS methods

    International Nuclear Information System (INIS)

    Kavetskyy, T.; Vakiv, M.; Shpotyuk, O.

    2007-01-01

    A combination of Raman scattering and positron annihilation lifetime spectroscopy (PALS) techniques to study charged defects in chalcogenide vitreous semiconductors (ChVSs) was applied for the first time in this study. In the case of Ge 15.8 As 21 S 63.2 glass, it is found that the main radiation-induced switching of heteropolar Ge-S bonds into heteropolar As-S ones, previously detected by IR fast Fourier transform spectroscopy, can also be identified by Raman spectroscopy in the depolarized configuration. Results obtained by Raman scattering are in good agreement with PALS data for the investigated glass composition

  19. Charged defects in chalcogenide vitreous semiconductors studied with combined Raman scattering and PALS methods

    Energy Technology Data Exchange (ETDEWEB)

    Kavetskyy, T.; Vakiv, M. [Lviv Institute of Materials of SRC ' Carat' , 202 Stryjska str., Lviv, UA-79031 (Ukraine); Shpotyuk, O. [Lviv Institute of Materials of SRC ' Carat' , 202 Stryjska str., Lviv, UA-79031 (Ukraine)], E-mail: shpotyuk@novas.lviv.ua

    2007-04-15

    A combination of Raman scattering and positron annihilation lifetime spectroscopy (PALS) techniques to study charged defects in chalcogenide vitreous semiconductors (ChVSs) was applied for the first time in this study. In the case of Ge{sub 15.8}As{sub 21}S{sub 63.2} glass, it is found that the main radiation-induced switching of heteropolar Ge-S bonds into heteropolar As-S ones, previously detected by IR fast Fourier transform spectroscopy, can also be identified by Raman spectroscopy in the depolarized configuration. Results obtained by Raman scattering are in good agreement with PALS data for the investigated glass composition.

  20. Recent highlights of the PALS research program

    Czech Academy of Sciences Publication Activity Database

    Jungwirth, Karel

    2005-01-01

    Roč. 23, - (2005), s. 177-182 ISSN 0263-0346. [ECLIM 2004: European Conference on Laser Interaction with Matter /28./. Roma, 06.09.2004-10.09.2004] R&D Projects: GA AV ČR(CZ) KSK2043105; GA MŠk(CZ) LN00A100 Grant - others:European Commission(XE) HPRI-CT-1999-00053 Institutional research plan: CEZ:AV0Z10100523 Keywords : ablation * Iodine lasers * laser ion sources * laser-produced plasma * laser-target interaction * PALS * shock waves * XUV lasers Subject RIV: BH - Optics, Masers, Lasers Impact factor: 2.590, year: 2005

  1. Activation of the jasmonic acid pathway by depletion of the hydroperoxide lyase OsHPL3 reveals crosstalk between the HPL and AOS branches of the oxylipin pathway in rice.

    Directory of Open Access Journals (Sweden)

    Xiaoqiang Liu

    Full Text Available The allene oxide synthase (AOS and hydroperoxide lyase (HPL branches of the oxylipin pathway, which underlie the production of jasmonates and aldehydes, respectively, function in plant responses to a range of stresses. Regulatory crosstalk has been proposed to exist between these two signaling branches; however, there is no direct evidence of this. Here, we identified and characterized a jasmonic acid (JA overproduction mutant, cea62, by screening a rice T-DNA insertion mutant library for lineages that constitutively express the AOS gene. Map-based cloning was used to identify the underlying gene as hydroperoxide lyase OsHPL3. HPL3 expression and the enzyme activity of its product, (E-2-hexenal, were depleted in the cea62 mutant, which resulted in the dramatic overproduction of JA, the activation of JA signaling, and the emergence of the lesion mimic phenotype. A time-course analysis of lesion formation and of the induction of defense responsive genes in the cea62 mutant revealed that the activation of JA biosynthesis and signaling in cea62 was regulated in a developmental manner, as was OsHPL3 activity in the wild-type plant. Microarray analysis showed that the JA-governed defense response was greatly activated in cea62 and this plant exhibited enhanced resistance to the T1 strain of the bacterial blight pathogen Xanthomonasoryzaepvoryzae (Xoo. The wounding response was attenuated in cea62 plants during the early stages of development, but partially recovered when JA levels were elevated during the later stages. In contrast, the wounding response was not altered during the different developmental stages of wild-type plants. These findings suggest that these two branches of the oxylipin pathway exhibit crosstalk with regards to biosynthesis and signaling and cooperate with each other to function in diverse stress responses.

  2. Pectinolytic bacteria and their secreted pectate lyases: agents for the maceration and solubilization of phytomass for fuels production

    Energy Technology Data Exchange (ETDEWEB)

    Preston, J.F. III; Rice, J.D.; Chow, M.C. (Florida Univ., Gainesville, FL (United States). Dept. of Microbiology and Cell Science)

    1993-01-01

    The objectives of this research have been to identify the pectinolytic enzymes secreted by bacteria and apply these towards the enhanced maceration and solubilization of plant material, focusing on the pectate lyases secreted by the phytopathogenic strains of Erwinia chrysanthemi, the ruminant resident Lachnospira multiparus, and the wood digestor isolate, Clostridium populeti. An HPLC approach has been developed that permits the kinetic analysis of each enzyme with respect to the formation of individual products during the pectate depolymerization process. This approach has demonstrated that each of these organisms secretes a nonrandom trimer-generating pectate lyase with a combination of endolytic and exolytic depolymerizing mechanisms. Two different strains of E. chrysanthemi secrete a battery of pectate lyases that include random endolytic as well as nonrandom dimer - and nonrandom trimer-generating endolytic/exolytic mechanisms. (author)

  3. Development and feasibility testing of an intervention to support active lifestyles in youths with type 1 diabetes-the ActivPals programme: a study protocol.

    Science.gov (United States)

    Mitchell, Fiona; Kirk, Alison; Robertson, Kenneth; Reilly, John J

    2016-01-01

    The global incidence of type 1 diabetes is rising, and youths with type 1 diabetes continue to suffer poorer health than peers without diabetes. Evidence suggests youths with type 1 diabetes have physical activity (PA) levels well below the recommendations for health and have high levels of sedentary behaviour. An active lifestyle is therefore recommended to improve health. There is limited research showing effective lifestyle behaviour change in this population; therefore, an evidence gap exists between the need to promote physical activity in type 1 diabetes care and lack of understanding on how to do this. This protocol paper describes a feasibility and pilot study of the ActivPals programme-an intervention to support active lifestyles in youths with type 1 diabetes. Key intervention components have been identified from preliminary work (individual and family focus, peer mentoring, technology integration and improved communication and understanding) and are being developed into a pragmatic randomised controlled trial (RCT) supported by recruitment pathways. A steering group of health care professionals and managers will refine the intervention to patient needs. A pilot trial is providing data on intervention implementation, acceptability and feasibility. Twenty youths with type 1 diabetes are being recruited and randomised into an intervention or control group. Physical activity is being measured objectively using the Actigraph GT3X+ monitor at baseline and 1-month follow-up. Contextual factors associated with intervention delivery are being explored. This study will contribute to the development of evidence-based, user-informed and pragmatic interventions leading to healthier lifestyles in youths with type 1 diabetes.

  4. Catalytic Mechanisms and Biocatalytic Applications of Aspartate and Methylaspartate Ammonia Lyases

    NARCIS (Netherlands)

    de Villiers, Marianne; Veetil, Vinod Puthan; Raj, Hans; de Villiers, Jandre; Poelarends, Gerrit J.

    2012-01-01

    Ammonia lyases catalyze the formation of alpha-beta-unsaturated bonds by the elimination of ammonia from their substrates. This conceptually straightforward reaction has been the emphasis of many studies, with the main focus on the catalytic mechanism of these enzymes and/or the use of these enzymes

  5. S1P lyase in thymic perivascular spaces promotes egress of mature thymocytes via up-regulation of S1P receptor 1.

    Science.gov (United States)

    Maeda, Yasuhiro; Yagi, Hideki; Takemoto, Kana; Utsumi, Hiroyuki; Fukunari, Atsushi; Sugahara, Kunio; Masuko, Takashi; Chiba, Kenji

    2014-05-01

    Sphingosine 1-phosphate (S1P) and S1P receptor 1 (S1P1) play an important role in the egress of mature CD4 or CD8 single-positive (SP) thymocytes from the thymus. Fingolimod hydrochloride (FTY720), an S1P1 functional antagonist, induced significant accumulation of CD62L(high)CD69(low) mature SP thymocytes in the thymic medulla. Immunohistochemical staining using anti-S1P1 antibody revealed that S1P1 is predominantly expressed on thymocytes in the thymic medulla and is strongly down-regulated even at 3h after FTY720 administration. 2-Acetyl-4-tetrahydroxybutylimidazole (THI), an S1P lyase inhibitor, also induced accumulation of mature SP thymocytes in the thymic medulla with an enlargement of the perivascular spaces (PVS). At 6h after THI administration, S1P1-expressing thymocytes reduced partially as if to form clusters and hardly existed in the proximity of CD31-expressing blood vessels in the thymic medulla, suggesting S1P lyase expression in the cells constructing thymic medullary PVS. To determine the cells expressing S1P lyase in the thymus, we newly established a mAb (YK19-2) specific for mouse S1P lyase. Immunohistochemical staining with YK19-2 revealed that S1P lyase is predominantly expressed in non-lymphoid thymic stromal cells in the thymic medulla. In the thymic medullary PVS, S1P lyase was expressed in ER-TR7-positive cells (reticular fibroblasts and pericytes) and CD31-positive vascular endothelial cells. Our findings suggest that S1P lyase expressed in the thymic medullary PVS keeps the tissue S1P concentration low around the vessels and promotes thymic egress via up-regulation of S1P1.

  6. Crystallization and preliminary X-ray analysis of argininosuccinate lyase from Streptococcus mutans

    International Nuclear Information System (INIS)

    Cao, Yan-Li; Li, Gui-Lan; Wang, Kai-Tuo; Zhang, Hong-Yin; Li, Lan-Fen

    2011-01-01

    Crystals of argininosuccinate lyase from S. mutans were obtained and X-ray data were collected to 2.5 Å resolution in space group R3. Argininosuccinate lyase (ASL) is an important enzyme in arginine synthesis and the urea cycle, which are highly conserved from bacteria to eukaryotes. The gene encoding Streptococcus mutans ASL (smASL) was amplified and cloned into expression vector pET28a. The recombinant smASL protein was expressed in a soluble form in Escherichia coli strain BL21 (DE3) and purified to homogeneity by two-step column chromatography. Crystals suitable for X-ray analysis were obtained and X-ray diffraction data were collected to a resolution of 2.5 Å. The crystals belonged to space group R3, with unit-cell parameters a = b = 254.5, c = 78.3 Å

  7. Plant growth-promoting Methylobacterium induces defense responses in groundnut (Arachis hypogaea L.) compared with rot pathogens.

    Science.gov (United States)

    Madhaiyan, M; Suresh Reddy, B V; Anandham, R; Senthilkumar, M; Poonguzhali, S; Sundaram, S P; Sa, Tongmin

    2006-10-01

    This study, framed in two different phases, studied the plant-growth promotion and the induction of systemic resistance in groundnut by Methylobacterium. Seed imbibition with Methylobacterium sp. increased germination by 19.5% compared with controls. Combined inoculation of Methylobacterium sp. with Rhizobium sp. also significantly increased plant growth, nodulation, and yield attributes in groundnut compared with individual inoculation of Rhizobium sp. Methylobacterium sp. challenge-inoculated with Aspergillus niger/Sclerotium rolfsii in groundnut significantly enhanced germination percentage and seedling vigour and showed increased phenylalanine ammonia lyase (PAL), beta-1,3-glucanase, and peroxidase (PO) activities. Under pot-culture conditions, in Methylobacterium sp. seed-treated groundnut plants challenge-inoculated with A. niger/S. rolfsii through foliar sprays on day 30, the activities of enzymes PO, PAL, and beta-1,3-glucanase increased constantly from 24 to 72 hours, after which decreased activity was noted. Five isozymes of polyphenol oxidase and PO could be detected in Methylobacterium-treated plants challenged with A. niger/S. rolfsii. Induced systemic resistance activity in groundnut against rot pathogens in response to methylotrophic bacteria suggests the possibility that pink-pigmented facultative methylotrophic bacteria might be used as a means of biologic disease control.

  8. Effect on isoflavone of soybean seedlings by 532nm laser irradiation

    Science.gov (United States)

    Tian, J.; Jin, L. H.; Li, J. M.; Shen, B. J.; Wang, C. Y.; Lu, X.; Zhao, X. L.

    2010-02-01

    We took soybeans as experimental substance to study how the 532nm laser with different power density and irradiation time affected the pullulation ratio of the soybeans, average height of seedlings and the isoflavone content of seedlings' cotyledon and laminae. The mechanism that laser pretreatment of soybean seeds could increase the isoflavone content of the seedlings was discussed in such both aspects as the efficiency of the photosynthesis and the activity of a phenylalanine ammonia-lyase (PAL) as an initial enzyme for synthesizing the isoflavone. The results showed that after the soybean seeds were pretreated by laser, the activity of the PAL and the resultants of the photosynthesis such as the sugar of dissolubility, the sucrose, and the amylum all increased with the soybean seeds irradiated by laser in which the effect on the soybean seeds pretreated by 15mW/mm2 laser for 5 min was the most obvious. As a result, the photosynthesis efficiency of the soybean seedlings increased after being pretreated by laser, which might offer the foundation for accumulating a large amount of isoflavone.

  9. A Catalase-related Hemoprotein in Coral Is Specialized for Synthesis of Short-chain Aldehydes: DISCOVERY OF P450-TYPE HYDROPEROXIDE LYASE ACTIVITY IN A CATALASE.

    Science.gov (United States)

    Teder, Tarvi; Lõhelaid, Helike; Boeglin, William E; Calcutt, Wade M; Brash, Alan R; Samel, Nigulas

    2015-08-07

    In corals a catalase-lipoxygenase fusion protein transforms arachidonic acid to the allene oxide 8R,9-epoxy-5,9,11,14-eicosatetraenoic acid from which arise cyclopentenones such as the prostanoid-related clavulones. Recently we cloned two catalase-lipoxygenase fusion protein genes (a and b) from the coral Capnella imbricata, form a being an allene oxide synthase and form b giving uncharacterized polar products (Lõhelaid, H., Teder, T., Tõldsepp, K., Ekins, M., and Samel, N. (2014) PloS ONE 9, e89215). Here, using HPLC-UV, LC-MS, and NMR methods, we identify a novel activity of fusion protein b, establishing its role in cleaving the lipoxygenase product 8R-hydroperoxy-eicosatetraenoic acid into the short-chain aldehydes (5Z)-8-oxo-octenoic acid and (3Z,6Z)-dodecadienal; these primary products readily isomerize in an aqueous medium to the corresponding 6E- and 2E,6Z derivatives. This type of enzymatic cleavage, splitting the carbon chain within the conjugated diene of the hydroperoxide substrate, is known only in plant cytochrome P450 hydroperoxide lyases. In mechanistic studies using (18)O-labeled substrate and incubations in H2(18)O, we established synthesis of the C8-oxo acid and C12 aldehyde with the retention of the hydroperoxy oxygens, consistent with synthesis of a short-lived hemiacetal intermediate that breaks down spontaneously into the two aldehydes. Taken together with our initial studies indicating differing gene regulation of the allene oxide synthase and the newly identified catalase-related hydroperoxide lyase and given the role of aldehydes in plant defense, this work uncovers a potential pathway in coral stress signaling and a novel enzymatic activity in the animal kingdom. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

  10. [Sudden death of a patient with 3-hydroxy-3-methylglutaryl coenzyme A lyase deficiency].

    Science.gov (United States)

    Vilaseca Busca, M A; Ribes Rubio, A; Briones Godino, P; Cusi Sánchez, V; Baraíbar Castelló, R; Gairi Taull, J M

    1990-02-01

    A new case of neonatal 3-hydroxy-3-methylglutaric aciduria is described. 3-hydroxy-3-methylglutaryl CoA lyase activities in leukocytes demonstrated the patient's homozygosity and the heterozygous character of the parents and two other members of the family. Dietetic management with low fat high carbohydrate diet together with protein restriction and carnitine resulted in a good control of the metabolic acidosis, the hypoglycemia, and the physical and neurological development. Nevertheless, sudden death occurred at the age thirteen months without any previous apparent trouble and the necropsia showed neither signs of infection nor hepatic or cardiac derangement.

  11. Engineering methylaspartate ammonia lyase for the asymmetric synthesis of unnatural amino acids

    NARCIS (Netherlands)

    Raj, Hans; Szymanski, Wiktor; de Villiers, Jandre; Rozeboom, Henriëtte J.; Puthan Veetil, Vinod; Reis, Carlos R.; Villiers, Marianne de; Dekker, Frank J.; Wildeman, Stefaan de; Quax, Wim J.; Thunnissen, Andy-Mark W.H.; Feringa, Ben L.; Janssen, Dick B.; Poelarends, Gerrit J.

    The redesign of enzymes to produce catalysts for a predefined transformation remains a major challenge in protein engineering. Here, we describe the structure-based engineering of methylaspartate ammonia lyase (which in nature catalyses the conversion of 3-methylaspartate to ammonia and

  12. ¹³C metabolic flux analysis identifies an unusual route for pyruvate dissimilation in mycobacteria which requires isocitrate lyase and carbon dioxide fixation.

    Directory of Open Access Journals (Sweden)

    Dany J V Beste

    2011-07-01

    Full Text Available Mycobacterium tuberculosis requires the enzyme isocitrate lyase (ICL for growth and virulence in vivo. The demonstration that M. tuberculosis also requires ICL for survival during nutrient starvation and has a role during steady state growth in a glycerol limited chemostat indicates a function for this enzyme which extends beyond fat metabolism. As isocitrate lyase is a potential drug target elucidating the role of this enzyme is of importance; however, the role of isocitrate lyase has never been investigated at the level of in vivo fluxes. Here we show that deletion of one of the two icl genes impairs the replication of Mycobacterium bovis BCG at slow growth rate in a carbon limited chemostat. In order to further understand the role of isocitrate lyase in the central metabolism of mycobacteria the effect of growth rate on the in vivo fluxes was studied for the first time using ¹³C-metabolic flux analysis (MFA. Tracer experiments were performed with steady state chemostat cultures of BCG or M. tuberculosis supplied with ¹³C labeled glycerol or sodium bicarbonate. Through measurements of the ¹³C isotopomer labeling patterns in protein-derived amino acids and enzymatic activity assays we have identified the activity of a novel pathway for pyruvate dissimilation. We named this the GAS pathway because it utilizes the Glyoxylate shunt and Anapleurotic reactions for oxidation of pyruvate, and Succinyl CoA synthetase for the generation of succinyl CoA combined with a very low flux through the succinate--oxaloacetate segment of the tricarboxylic acid cycle. We confirm that M. tuberculosis can fix carbon from CO₂ into biomass. As the human host is abundant in CO₂ this finding requires further investigation in vivo as CO₂ fixation may provide a point of vulnerability that could be targeted with novel drugs. This study also provides a platform for further studies into the metabolism of M. tuberculosis using ¹³C-MFA.

  13. Changes in polyphenol profile of dried apricots containing SO2 at various concentrations during storage.

    Science.gov (United States)

    Altındağ, Melek; Türkyılmaz, Meltem; Özkan, Mehmet

    2018-05-01

    Changes in polyphenols have important effects on the quality (especially color) and health benefits of dried apricots. SO 2 concentration, storage and the activities of polyphenol oxidase (PPO) and phenylalanine ammonia lyase (PAL) were factors which had significant effects on polyphenols. Polyphenol profile and activities of PPO and PAL in sulfured dried apricots (SDAs, 0, 451, 832, 2112 and 3241 mg SO 2 kg -1 ) were monitored during storage at 4, 20 and 30 °C for 379 days for the first time. Even the lowest SO 2 concentration (451 mg kg -1 ) was sufficient to inactivate PPO during the entire storage period. However, while SO 2 led to the increase in PAL activity of the samples (r = 0.767) before storage, PAL activities of SDAs decreased during storage. After 90 days of storage, PAL activity was determined in only non-sulfured dried apricots (NSDAs) and dried apricots containing 451 mg SO 2 kg -1 . Although the major polyphenol in NSDAs was epicatechin (611.4 mg kg -1 ), that in SDAs was chlorogenic acid (455-1508 mg kg -1 ), followed by epicatechin (0-426.8 mg kg -1 ), rutin (148.9-477.3 mg kg -1 ), ferulic acid (23.3-55.3 mg kg -1 ) and gallic acid (2.4-43.6 mg kg -1 ). After storage at 30 °C for 379 days, the major polyphenol in SDAs was gallic acid (706-2324 mg kg -1 ). However, the major polyphenol in NSDAs did not change after storage. The highest total polyphenol content was detected in SDAs containing 2112 mg SO 2 kg -1 and stored at 30 °C. To produce dried apricots having high polyphenol content, ∼2000 mg SO 2 kg -1 should be used. Low storage temperature (<30 °C) was not necessary for the protection of polyphenols. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

  14. Clam-Gallasův palác - emblematická architektura?

    Czech Academy of Sciences Publication Activity Database

    Konečný, Lubomír

    2009-01-01

    Roč. 28, - (2009), s. 91-98 ISSN 0231-7443. [Život pražských paláců. Praha, 09.10.2007-11.10.2007] Institutional research plan: CEZ:AV0Z80330511 Keywords : Clam-Gallas Palace * Johann Bernhard Fischer von Erlach * architecture * emblem s * imprints Subject RIV: AL - Art, Architecture, Cultural Heritage

  15. Characterization of the TolB-Pal trans-envelope complex from Xylella fastidiosa reveals a dynamic and coordinated protein expression profile during the biofilm development process.

    Science.gov (United States)

    Santos, Clelton A; Janissen, Richard; Toledo, Marcelo A S; Beloti, Lilian L; Azzoni, Adriano R; Cotta, Monica A; Souza, Anete P

    2015-10-01

    The intriguing roles of the bacterial Tol-Pal trans-envelope protein complex range from maintenance of cell envelope integrity to potential participation in the process of cell division. In this study, we report the characterization of the XfTolB and XfPal proteins of the Tol-Pal complex of Xylella fastidiosa. X. fastidiosa is a major plant pathogen that forms biofilms inside xylem vessels, triggering the development of diseases in important cultivable plants around the word. Based on functional complementation experiments in Escherichia coli tolB and pal mutant strains, we confirmed the role of xftolB and xfpal in outer membrane integrity. In addition, we observed a dynamic and coordinated protein expression profile during the X. fastidiosa biofilm development process. Using small-angle X-ray scattering (SAXS), the low-resolution structure of the isolated XfTolB-XfPal complex in solution was solved for the first time. Finally, the localization of the XfTolB and XfPal polar ends was visualized via immunofluorescence labeling in vivo during bacterial cell growth. Our results highlight the major role of the components of the cell envelope, particularly the TolB-Pal complex, during the different phases of bacterial biofilm development. Copyright © 2015 Elsevier B.V. All rights reserved.

  16. Change in the Content of Salicylic Acid and in the Activities of Phenylalanine Ammonia-Lyase and Catalase in Wheat Seedling Roots Under the Effect of Azospirillum Lectins

    Directory of Open Access Journals (Sweden)

    Alen'kina S.A.

    2012-05-01

    Full Text Available We investigated the time course of changes in the endogenous content of salicylic acid, the ratio between the acid's free and bound forms, and changes in the activities of phenylalanine ammonia-lyase and catalase in wheat seedling roots under the effect of the lectins of two strains of the associative nitrogen-fixing bacterium Azospirillum: A. brasilense Sp7 and its mutant defective in lectin activity, A. brasilense Sp7.2.3. Differences in plant response to the action of the lectins from these two strains were established. On the basis of the obtained data, a model was proposed for lectin-assisted induction of resistance, according to which the lectin effect on the roots of seedlings results in accumulation of free salicylic acid, which inhibits catalase activity, ultimately leading to accumulation of hydrogen peroxide and to formation of induced resistance.

  17. Genetic and molecular analyses of Escherichia coli N-acetylneuraminate lyase gene.

    OpenAIRE

    Kawakami, B; Kudo, T; Narahashi, Y; Horikoshi, K

    1986-01-01

    Two plasmids containing the N-acetylneuraminate lyase (NALase) gene (nanA) of Escherichia coli, pNL1 and pNL4, were constructed. Immunoprecipitation analysis indicated that the 35,000-dalton protein encoded in pNL4 was NALase. The synthesis of NALase in E. coli carrying these plasmids was constitutive.

  18. Alleviation of isoproturon toxicity to wheat by exogenous application of glutathione.

    Science.gov (United States)

    Nemat Alla, Mamdouh M; Hassan, Nemat M

    2014-06-01

    Treatment with the recommended field dose of isoproturon to 7-d-old wheat seedlings significantly decreased shoot height, fresh and dry weights during the subsequent 15days. Meanwhile contents of carotenoids, chlorophylls and anthocyanin as well as activities of δ-aminolevulinate dehydratase (ALA-D), phenylalanine ammonia lyase (PAL) and tyrosine ammonia lyase (TAL) were significantly inhibited. On the other hand, the herbicide significantly increased malondialdehyde (MDA), a naturally occurring product of lipid peroxidation and H2O2, while it significantly decreased the contents of glutathione (GSH) and ascorbic acid (AsA) and reduced the activities of superoxide dismutase (SOD), catalase (CAT) and ascorbate peroxidase (APX). These findings indicate an induction of a stress status in wheat seedlings following isoproturon treatment. However, exogenous GSH appeared to limit the toxic effects of isoproturon and seemed to overcome this stress status. Most likely, contents of pigment and activities of enzymes were raised to approximate control levels. Moreover, antioxidants were elevated and the oxidative stress indices seemed to be alleviated by GSH application. These results indicate that exogenous GSH enhances enzymatic and nonenzymatic antioxidants to alleviate the effects of isoproturon. Copyright © 2014 Elsevier Inc. All rights reserved.

  19. A horizontal gene transfer at the origin of phenylpropanoid metabolism: a key adaptation of plants to land.

    Science.gov (United States)

    Emiliani, Giovanni; Fondi, Marco; Fani, Renato; Gribaldo, Simonetta

    2009-02-16

    The pioneering ancestor of land plants that conquered terrestrial habitats around 500 million years ago had to face dramatic stresses including UV radiation, desiccation, and microbial attack. This drove a number of adaptations, among which the emergence of the phenylpropanoid pathway was crucial, leading to essential compounds such as flavonoids and lignin. However, the origin of this specific land plant secondary metabolism has not been clarified. We have performed an extensive analysis of the taxonomic distribution and phylogeny of Phenylalanine Ammonia Lyase (PAL), which catalyses the first and essential step of the general phenylpropanoid pathway, leading from phenylalanine to p-Coumaric acid and p-Coumaroyl-CoA, the entry points of the flavonoids and lignin routes. We obtained robust evidence that the ancestor of land plants acquired a PAL via horizontal gene transfer (HGT) during symbioses with soil bacteria and fungi that are known to have established very early during the first steps of land colonization. This horizontally acquired PAL represented then the basis for further development of the phenylpropanoid pathway and plant radiation on terrestrial environments. Our results highlight a possible crucial role of HGT from soil bacteria in the path leading to land colonization by plants and their subsequent evolution. The few functional characterizations of sediment/soil bacterial PAL (production of secondary metabolites with powerful antimicrobial activity or production of pigments) suggest that the initial advantage of this horizontally acquired PAL in the ancestor of land plants might have been either defense against an already developed microbial community and/or protection against UV.

  20. Fast ion emission from the plasma produced by the PALS laser system

    Czech Academy of Sciences Publication Activity Database

    Wolowski, J.; Badziak, J.; Boody, F. P.; Hora, H.; Hnatowicz, Vladimír; Jungwirth, Karel; Krása, Josef; Láska, Leoš; Parys, P.; Peřina, Vratislav; Pfeifer, Miroslav; Rohlena, Karel; Ryc, L.; Ullschmied, Jiří; Woryna, E.

    2002-01-01

    Roč. 44, - (2002), s. 1277-1283 ISSN 0741-3335 Institutional research plan: CEZ:AV0Z1048901 Keywords : emission * plasma produced * PALS laser system ? Subject RIV: BG - Nuclear, Atomic and Molecular Physics, Colliders Impact factor: 2.121, year: 2002

  1. Feasibility and reliability of remote assessment of PALS psychomotor skills via interactive videoconferencing.

    Science.gov (United States)

    Weeks, Douglas L; Molsberry, Dianne M

    2009-03-01

    This study determined inter-rater agreement between skill assessments provided by on-site PALS evaluators with ratings from evaluators at a remote site viewing the same skill performance over a videoconferencing network. Judgments about feasibility of remote evaluation were also obtained from the evaluators and PALS course participants. Two remote and two on-site instructors independently rated performance of 27 course participants who performed cardiac and shock/respiratory emergency core cases. Inter-rater reliability was assessed with the intraclass correlation coefficient (ICC). Feasibility was assessed with surveys of evaluators and course participants. Core cases were under the direction of the remote evaluators. The ICC for overall agreement on pass/fail decisions was 0.997 for the cardiac cases and 0.998 for the shock/respiratory cases. Perfect agreement was reached on 52 of 54 pass/fail decisions. Across all evaluators, all core cases, and all participants, 2584 ratings of individual skill criteria were provided, of which 21 (0.8%) were ratings in which a single evaluator disagreed with the other three evaluators. No trends emerged for location of the disagreeing evaluator. Survey responses indicated that remote evaluation was acceptable and feasible to course participants and to the evaluators. Videoconferencing technology was shown to provide adequate spatial and temporal resolution for PALS evaluators at-a-distance from course participants to agree with ratings of on-site evaluators.

  2. Optimisation of trans-cinnamic acid and hydrocinnamyl alcohol production with recombinant Saccharomyces cerevisiae and identification of cinnamyl methyl ketone as a by-product.

    Science.gov (United States)

    Gottardi, Manuela; Grün, Peter; Bode, Helge B; Hoffmann, Thomas; Schwab, Wilfried; Oreb, Mislav; Boles, Eckhard

    2017-12-01

    Trans-cinnamic acid (tCA) and hydrocinnamyl alcohol (HcinOH) are valuable aromatic compounds with applications in the flavour, fragrance and cosmetic industry. They can be produced with recombinant yeasts from sugars via phenylalanine after expression of a phenylalanine ammonia lyase (PAL) and an aryl carboxylic acid reductase. Here, we show that in Saccharomyces cerevisiae a PAL enzyme from the bacterium Photorhabdus luminescens was superior to a previously used plant PAL enzyme for the production of tCA. Moreover, after expression of a UDP-glucose:cinnamate glucosyltransferase (FaGT2) from Fragaria x ananassa, tCA could be converted to cinnamoyl-D-glucose which is expected to be less toxic to the yeast cells. Production of tCA and HcinOH from glucose could be increased by eliminating feedback-regulated steps of aromatic amino acid biosynthesis and diminishing the decarboxylation step of the competing Ehrlich pathway. Finally, an unknown by-product resulting from further metabolisation of a carboligation product of cinnamaldehyde (cinALD) with activated acetaldehyde, mediated by pyruvate decarboxylases, could be identified as cinnamyl methyl ketone providing a new route for the biosynthesis of precursors, such as (2S,3R) 5-phenylpent-4-ene-2,3-diol, necessary for the chemical synthesis of specific biologically active drugs such as daunomycin. © FEMS 2017. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  3. Stress responses of duckweed (Lemna minor L.) and water velvet (Azolla filiculoides Lam.) to anionic surfactant sodium-dodecyl-sulphate (SDS).

    Science.gov (United States)

    Forni, C; Braglia, R; Harren, F J M; Cristescu, S M

    2012-04-01

    Surfactants are used for several purposes and recently they have attracted the attention for their ability to modify the behavior of other preexistent or co-disposed contaminants, although their use or discharge in wastewaters can represent a real or potential risk for the environment. Lemna minor L. and Azolla filiculoides Lam. are floating aquatic macrophytes, very effective in accumulating several pollutants including sodium dodecyl sulphate (SDS). In this work we evaluated the effects of SDS on these species by determining the stress ethylene production via laser-based trace gas detection, and the activities of enzymes involved in stress response, such as guaiacol peroxidase (G-POD), phenylalanine ammonia-lyase (PAL) and polyphenol-oxidase (PPO). Phenolics content was also determined. The macrophytes were treated with different concentrations of SDS for one week. SDS affected duckweed enzymatic activities and phenol content. While in the fern phenolics amount, PAL, G-POD and PPO activities were not affected by SDS except for 100 ppm SDS, the only concentration that was taken up and not completely degraded. Stress ethylene production was induced only in the fern treated with 50 and 100 ppm SDS. Copyright © 2011 Elsevier B.V. All rights reserved.

  4. Perturbation of formate pathway for hydrogen production by expressions of formate hydrogen lyase and its transcriptional activator in wild Enterobacter aerogenes and its mutants

    Energy Technology Data Exchange (ETDEWEB)

    Lu, Yuan; Zhao, Hongxin; Zhang, Chong; Lai, Qiheng; Xing, Xin-Hui [Institute of Biochemical Engineering, Department of Chemical Engineering, Tsinghua University, Beijing 100084 (China)

    2009-06-15

    To examine perturbation effects of formate pathway on hydrogen productivity in Enterobacter aerogenes (Ea), formate dehydrogenase FDH-H gene (fdhF) and formate hydrogen lyase activator protein FHLA gene (fhlA) originated from Escherichia coli, were overexpressed in the wild strain Ea, its hycA-deleted mutant (A) by knockout the formate hydrogen lyase repressor and hybO-deleted mutant (O) by knockout of the uptake hydrogenase, respectively. Overexpression of fdhF and fhlA promoted cell growth and volumetric hydrogen production rates of all the strains, and the hydrogen production per gram cell dry weight (CDW) for Ea, A and O was increased by 38.5%, 21.8% and 5.25%, respectively. The fdhF and fhlA overexpression improved the hydrogen yield per mol glucose of strains Ea and A, but declined that of strain O. The increase of hydrogen yield of the strain Ea with fdhF and fhlA expression was mainly attributed to the increase of formate pathway, while for the mutant A, the improved hydrogen yield with fdhF and fhlA expression was mainly due to the increase of NADH pathway. Analysis of the metabolites and ratio of ethanol-to-acetate showed that the cellular redox state balance and energy level were also changed for these strains by fdhF and fhlA expression. These findings demonstrated that the hydrogen production was not only dependent on the hydrogenase genes, but was also affected by the regulation of the whole metabolism. Therefore, fdhF and fhlA expression in different strains of E. aerogenes could exhibit different perturbation effects on the metabolism and the hydrogen productivity. (author)

  5. Water-Vapor Sorption Processes in Nanoporous MgO-Al2O3 Ceramics: the PAL Spectroscopy Study

    OpenAIRE

    Klym, Halyna; Ingram, Adam; Shpotyuk, Oleh; Hadzaman, Ivan; Solntsev, Viacheslav

    2016-01-01

    The water-vapor sorption processes in nanoporous MgO-Al2O3 ceramics are studied with positron annihilation lifetime (PAL) spectroscopy employing positron trapping and positronium (Ps)-decaying modes. It is demonstrated that the longest-lived components in the four-term reconstructed PAL spectra with characteristic lifetimes near 2 and 60?70?ns can be, respectively, attributed to ortho-positronium (o-Ps) traps in nanopores with 0.3- and 1.5?1.8-nm radii. The first o-Ps decaying process include...

  6. The Skin Bacterium Propionibacterium acnes Employs Two Variants of Hyaluronate Lyase with Distinct Properties

    DEFF Research Database (Denmark)

    Nazipi, Seven; Stødkilde-Jørgensen, Kristian; Scavenius, Carsten

    2017-01-01

    Hyaluronic acid (HA) and other glycosaminoglycans are extracellular matrix components in the human epidermis and dermis. One of the most prevalent skin microorganisms, Propionibacterium acnes, possesses HA-degrading activity, possibly conferred by the enzyme hyaluronate lyase (HYL). In this study......, we identified the HYL of P. acnes and investigated the genotypic and phenotypic characteristics. Investigations include the generation of a P. acneshyl knockout mutant and HYL activity assays to determine the substrate range and formed products. We found that P. acnes employs two distinct variants...... of the observed differences between P. acnes phylotype IA and IB/II strains. Whereas type IA strains are primarily found on the skin surface and associated with acne vulgaris, type IB/II strains are more often associated with soft and deep tissue infections, which would require elaborate tissue invasion...

  7. Endogenous salicylic acid shows different correlation with baicalin and baicalein in the medicinal plant Scutellaria baicalensis Georgi subjected to stress and exogenous salicylic acid.

    Directory of Open Access Journals (Sweden)

    Hu Su

    Full Text Available Salicylic acid (SA is synthesized via the phenylalanine lyase (PAL and isochorismate synthase (ICS pathways and can influence the stress response in plants by regulating certain secondary metabolites. However, the association between SA and particular secondary metabolites in the Chinese medicinal plant Scutellaria baicalensis Georgi is unclear. To elucidate the association between SA and the secondary metabolites baicalin and baicalein, which constitute the primary effective components of S. baicalensis, we subjected seedlings to drought and salt stress and exogenous SA treatment in a laboratory setting and tested the expression of PAL and ICS, as well as the content of free SA (FSA, total SA (TSA, baicalin, and baicalein. We also assessed the correlation of FSA and TSA with PAL and ICS, and with baicalin and baicalein accumulation, respectively. The results indicated that both FSA and TSA were positively correlated with PAL, ICS, and baicalin, but negatively correlated with baicalein. The findings of this study improve our understanding of the manner in which SA regulates secondary metabolites in S. baicalensis.

  8. Gender differences in physical activity, sedentary behavior, and their relation to body composition in active Brazilian adolescents.

    Science.gov (United States)

    Nogueira, Julia Aparecida Devide; Macedo da Costa, Teresa Helena

    2009-01-01

    Body weight and composition are determined by genotype, environment, and energy balance. Physical activity or sedentary behavior have different associations with body weight, fat mass, and fat-free mass, a relationship that is not clear in adolescents. The aim of this study was to test the associations between gender, physical activity, sedentary behavior, and body composition in physically active adolescents. Weight, height, and skinfold thickness were measured in 326 physically active boys and girls age 11 to 15 years. All subjects answered a questionnaire assessing their usual daily activities for the last month. Time spent on each activity was used to estimate the physical activity level (PAL). PAL was associated with body composition after adjustment for age and maturation, with differences between genders. For boys, PAL was positively and significantly associated with body mass index (BMI) and fat-free mass index (beta=0.14 and 0.15, respectively). For girls, PAL was negatively and significantly associated with BMI and fat mass index (beta=-0.11 and -0.75, respectively). Sedentary behavior, expressed by hours of TV, videogame, and computer use, was not associated with any body-composition outcome for either gender. The accumulated amount of physical activity, but not of sedentary behavior, was related to body composition in active adolescents.

  9. Production of endo-pectate lyase by two stage cultivation of Erwinia carotovora

    Energy Technology Data Exchange (ETDEWEB)

    Fukuoka, Satoshi; Kobayashi, Yoshiaki

    1987-02-26

    The productivity of endo-pectate lyase from Erwinia carotovora GIR 1044 was found to be greatly improved by two stage cultivation: in the first stage the bacterium was grown with an inducing carbon source, e.g., pectin, and in the second stage it was cultivated with glycerol, xylose, or fructose with the addition of monosodium L-glutamate as nitrogen source. In the two stage cultivation using pectin or glycerol as the carbon source the enzyme activity reached 400 units/ml, almost 3 times as much as that of one stage cultivation in a 10 liter fermentor. Using two stage cultivation in the 200 liter fermentor improved enzyme productivity over that in the 10 liter fermentor, with 500 units/ml of activity. Compared with the cultivation in Erlenmeyer flasks, fermentor cultivation improved enzyme productivity. The optimum cultivating conditions were agitation of 480 rpm with aeration of 0.5 vvm at 28 /sup 0/C. (4 figs, 4 tabs, 14 refs)

  10. An acidic pectin lyase from Aspergillus niger with favourable efficiency in fruit juice clarification.

    Science.gov (United States)

    Xu, S X; Qin, X; Liu, B; Zhang, D Q; Zhang, W; Wu, K; Zhang, Y H

    2015-02-01

    The pectin lyase gene pnl-zj5a from Aspergillus niger ZJ5 was identified and expressed in Pichia pastoris. PNL-ZJ5A was purified by ultrafiltration, anion exchange and gel chromatography. The Km and Vmax values determined using citrus pectin were 0.66 mg ml(-1) and 32.6 μmol min(-1) mg(-1) , respectively. PNL-ZJ5A exhibited optimal activity at 43°C and retained activity over 25-50°C. PNL-ZJ5A was optimally active at pH 5 and effective in apple juice clarification. Compared with controls, PNL-ZJ5A increased the fruit juice yield significantly. Furthermore, PNL-ZJ5A reduced the viscosity of apple juice by 38.8% and increased its transmittance by 86.3%. PNL-ZJ5A combined with a commercial pectin esterase resulted in higher juice volume. © 2014 The Society for Applied Microbiology.

  11. Development and Evaluation of vetPAL, a Student-Led, Peer-Assisted Learning Program.

    Science.gov (United States)

    Bates, Lucy S W; Warman, Sheena; Pither, Zoe; Baillie, Sarah

    Based on an idea from a final-year student, Bristol Veterinary School introduced vetPAL, a student-led, peer-assisted learning program. The program involved fifth-year (final-year) students acting as tutors and leading sessions for fourth-year students (tutees) in clinical skills and revision (review) topics. The initiative aimed to supplement student learning while also providing tutors with opportunities to further develop a range of skills. All tutors received training and the program was evaluated using questionnaires collected from tutees and tutors after each session. Tutees' self-rated confidence increased significantly in clinical skills and for revision topics. Advantages of being taught by students rather than staff included the informal atmosphere, the tutees' willingness to ask questions, and the relatability of the tutors. The small group size and the style of learning in the revision sessions (i.e., group work, discussions, and interactivity) were additional positive aspects identified by both tutees and tutors. Benefits for tutors included developing their communication and teaching skills. The training sessions were considered key in helping tutors feel prepared to lead sessions, although the most difficult aspects were the lack of teaching experience and time management. Following the successful pilot of vetPAL, plans are in place to make the program permanent and sustainable, while incorporating necessary changes based on the evaluation and the student leader's experiences running the program. A vetPAL handbook has been created to facilitate organization of the program for future years.

  12. Phosphate enhances levan production in the endophytic bacterium Gluconacetobacter diazotrophicus Pal5

    Science.gov (United States)

    Idogawa, Nao; Amamoto, Ryuta; Murata, Kousaku; Kawai, Shigeyuki

    2014-01-01

    Gluconacetobacter diazotrophicus is a gram-negative and endophytic nitrogen-fixing bacterium that has several beneficial effects in host plants; thus, utilization of this bacterium as a biofertilizer in agriculture may be possible. G. diazotrophicus synthesizes levan, a D-fructofuranosyl polymer with β-(2→6) linkages, as an exopolysaccharide and the synthesized levan improves the stress tolerance of the bacterium. In this study, we found that phosphate enhances levan production by G. diazotrophicus Pal5, a wild type strain that showed a stronger mucous phenotype on solid medium containing 28 mM phosphate than on solid medium containing 7 mM phosphate. A G. diazotrophicus Pal5 levansucrase disruptant showed only a weak mucous phenotype regardless of the phosphate concentration, indicating that the mucous phenotype observed on 28 mM phosphate medium was caused by levan. To our knowledge, this is the first report of the effect of a high concentration of phosphate on exopolysaccharide production. PMID:24717418

  13. Biocatalytic Enantioselective Synthesis of N-Substituted Aspartic Acids by Aspartate Ammonia Lyase

    NARCIS (Netherlands)

    Weiner, Barbara; Poelarends, Gerrit J.; Janssen, Dick B.; Feringa, Ben L.

    2008-01-01

    The gene encoding aspartate ammonia lyase (aspB) from Bacillus sp. YM55-1 has been cloned and overexpressed, and the recombinant enzyme containing a C-terminal His6 tag has been purified to homogeneity and subjected to kinetic characterization. Kinetic studies have shown that the His6 tag does not

  14. Study of RNA interference inhibiting rat ovarian androgen biosynthesis by depressing 17alpha-hydroxylase/17, 20-lyase activity in vivo

    Directory of Open Access Journals (Sweden)

    Yang Xing

    2009-07-01

    Full Text Available Abstract Background 17alpha-hydroxylase/17, 20-lyase encoded by CYP17 is the key enzyme in androgen biosynthesis pathway. Previous studies demonstrated the accentuation of the enzyme in patients with polycystic ovary syndrome (PCOS was the most important mechanism of androgen excess. We chose CYP17 as the therapeutic target, trying to suppress the activity of 17alpha-hydroxylase/17, 20-lyase and inhibit androgen biosynthesis by silencing the expression of CYP17 in the rat ovary. Methods Three CYP17-targeting and one negative control oligonucleotides were designed and used in the present study. The silence efficiency of lentivirus shRNA was assessed by qRT-PCR, Western blotting and hormone assay. After subcapsular injection of lentivirus shRNA in rat ovary, the delivery efficiency was evaluated by GFP fluorescence and qPCR. Total RNA was extracted from rat ovary for CYP17 mRNA determination and rat serum was collected for hormone measurement. Results In total, three CYP17-targeting lentivirus shRNAs were synthesized. The results showed that all of them had a silencing effect on CYP17 mRNA and protein. Moreover, androstenedione secreted by rat theca interstitial cells (TIC in the RNAi group declined significantly compared with that in the control group. Two weeks after rat ovarian subcapsular injection of chosen CYP17 shRNA, the GFP fluorescence of frozen ovarian sections could be seen clearly under fluorescence microscope. It also showed that the GFP DNA level increased significantly, and its relative expression level was 7.42 times higher than that in the control group. Simultaneously, shRNA treatment significantly decreased CYP17 mRNA and protein levels at 61% and 54%, respectively. Hormone assay showed that all the levels of androstenedione, 17-hydroxyprogesterone and testosterone declined to a certain degree, but progesterone levels declined significantly. Conclusion The present study proves for the first time that ovarian androgen

  15. Requirement of argininosuccinate lyase for systemic nitric oxide production.

    Science.gov (United States)

    Erez, Ayelet; Nagamani, Sandesh C S; Shchelochkov, Oleg A; Premkumar, Muralidhar H; Campeau, Philippe M; Chen, Yuqing; Garg, Harsha K; Li, Li; Mian, Asad; Bertin, Terry K; Black, Jennifer O; Zeng, Heng; Tang, Yaoping; Reddy, Anilkumar K; Summar, Marshall; O'Brien, William E; Harrison, David G; Mitch, William E; Marini, Juan C; Aschner, Judy L; Bryan, Nathan S; Lee, Brendan

    2011-11-13

    Nitric oxide (NO) is crucial in diverse physiological and pathological processes. We show that a hypomorphic mouse model of argininosuccinate lyase (encoded by Asl) deficiency has a distinct phenotype of multiorgan dysfunction and NO deficiency. Loss of Asl in both humans and mice leads to reduced NO synthesis, owing to both decreased endogenous arginine synthesis and an impaired ability to use extracellular arginine for NO production. Administration of nitrite, which can be converted into NO in vivo, rescued the manifestations of NO deficiency in hypomorphic Asl mice, and a nitric oxide synthase (NOS)-independent NO donor restored NO-dependent vascular reactivity in humans with ASL deficiency. Mechanistic studies showed that ASL has a structural function in addition to its catalytic activity, by which it contributes to the formation of a multiprotein complex required for NO production. Our data demonstrate a previously unappreciated role for ASL in NOS function and NO homeostasis. Hence, ASL may serve as a target for manipulating NO production in experimental models, as well as for the treatment of NO-related diseases.

  16. Study of micro-phase separation of two polystyrene-based copolymer mixture using the combination of PALS and FT-IR

    International Nuclear Information System (INIS)

    Jiang, Z.Y.; Jiang, X.Q.; Yang, Y.X.; Huang, Y.J.; Huang, H.B.; Hsia, Y.F.

    2005-01-01

    Positron annihilation lifetime (PAL) spectroscopy, Fourier transform infrared (FT-IR) and differential scanning calorimetry (DSC) have been applied to study the micro-phase separation in the blends of poly(styrene-co-methylmethacrylate) (SMMA) copolymer and poly(styrene-co-maleic anhydride) (SMA) copolymer. The DSC results indicate that the SMA/SMMA blends are miscible and weak intermolecular interactions exist between SMA and SMMA. The strength of intermolecular interactions to some degree exhibits somewhat non-monotonic variation with increasing of SMA component in the blends. The results of PAL measurement present the blend containing 20 wt% SMA is phase-separated in molecular level, which is interpreted by the results of FT-IR analysis. It was concluded that it is helpful to study the miscibility of polymer blends in molecular level by means of PAL method, accompanied with the requisite measurement of DSC and FT-IR

  17. Refeeding syndrome in a young woman with argininosuccinate lyase deficiency.

    Science.gov (United States)

    Stuy, M; Chen, G-F; Masonek, J M; Scharschmidt, B F

    2015-09-01

    A severely chronically protein and calorie restricted young woman with argininosuccinate lyase deficiency developed transient refeeding syndrome (RFS) and hyperammonemia after modest diet liberalization following initiation of glycerol phenylbutyrate (GPB). The patient required IV supportive care and supplementation with potassium, magnesium and calcium. She is now doing well on GPB and an appropriate maintenance diet. Susceptibility to RFS should be considered in chronically nutritionally restricted patients with metabolic disorders after liberalization of diet.

  18. Refeeding syndrome in a young woman with argininosuccinate lyase deficiency☆

    Science.gov (United States)

    Stuy, M.; Chen, G.-F.; Masonek, J.M.; Scharschmidt, B.F.

    2015-01-01

    A severely chronically protein and calorie restricted young woman with argininosuccinate lyase deficiency developed transient refeeding syndrome (RFS) and hyperammonemia after modest diet liberalization following initiation of glycerol phenylbutyrate (GPB). The patient required IV supportive care and supplementation with potassium, magnesium and calcium. She is now doing well on GPB and an appropriate maintenance diet. Susceptibility to RFS should be considered in chronically nutritionally restricted patients with metabolic disorders after liberalization of diet. PMID:26937403

  19. From the Last Interglacial to the Anthropocene: Modelling a Complete Glacial Cycle (PalMod)

    Science.gov (United States)

    Brücher, Tim; Latif, Mojib

    2017-04-01

    We will give a short overview and update on the current status of the national climate modelling initiative PalMod (Paleo Modelling, www.palmod.de). PalMod focuses on the understanding of the climate system dynamics and its variability during the last glacial cycle. The initiative is funded by the German Federal Ministry of Education and Research (BMBF) and its specific topics are: (i) to identify and quantify the relative contributions of the fundamental processes which determined the Earth's climate trajectory and variability during the last glacial cycle, (ii) to simulate with comprehensive Earth System Models (ESMs) the climate from the peak of the last interglacial - the Eemian warm period - up to the present, including the changes in the spectrum of variability, and (iii) to assess possible future climate trajectories beyond this century during the next millennia with sophisticated ESMs tested in such a way. The research is intended to be conducted over a period of 10 years, but with shorter funding cycles. PalMod kicked off in February 2016. The first phase focuses on the last deglaciation (app. the last 23.000 years). From the ESM perspective PalMod pushes forward model development by coupling ESM with dynamical ice sheet models. Computer scientists work on speeding up climate models using different concepts (like parallelisation in time) and one working group is dedicated to perform a comprehensive data synthesis to validate model performance. The envisioned approach is innovative in three respects. First, the consortium aims at simulating a full glacial cycle in transient mode and with comprehensive ESMs which allow full interactions between the physical and biogeochemical components of the Earth system, including ice sheets. Second, we shall address climate variability during the last glacial cycle on a large range of time scales, from interannual to multi-millennial, and attempt to quantify the relative contributions of external forcing and processes

  20. The Phenylpropanoid Pathway and Lignin in Defense against Ganoderma boninense Colonized Root Tissues in Oil Palm (Elaeis guineensis Jacq.

    Directory of Open Access Journals (Sweden)

    Nisha T. Govender

    2017-08-01

    Full Text Available Basal stem rot, caused by the basidiomycete fungus, Ganoderma boninense, is an economically devastating disease in Malaysia. Our study investigated the changes in lignin content and composition along with activity and expression of the phenylpropanoid pathway enzymes and genes in oil palm root tissues during G. boninense infection. We sampled control (non-inoculated and infected (inoculated seedlings at seven time points [1, 2, 3, 4, 8, and 12 weeks post-inoculation (wpi] in a randomized design. The expression profiles of phenylalanine ammonia lyase (PAL, cinnamyl alcohol dehydrogenase (CAD, and peroxidase (POD genes were monitored at 1, 2, and 3 wpi using real-time quantitative polymerase chain reaction. Seedlings at 4, 8, and 12 wpi were screened for lignin content, lignin composition, enzyme activities (PAL, CAD, and POD, growth (weight and height, and disease severity (DS. Gene expression analysis demonstrated up-regulation of PAL, CAD, and POD genes in the infected seedlings, relative to the control seedlings at 1, 2, and 3 wpi. At 2 and 3 wpi, CAD showed highest transcript levels compared to PAL and POD. DS increased progressively throughout sampling, with 5, 34, and 69% at 4, 8, and 12 wpi, respectively. Fresh weight and height of the infected seedlings were significantly lower compared to the control seedlings at 8 and 12 wpi. Lignin content of the infected seedlings at 4 wpi was significantly higher than the control seedlings, remained elicited with no change at 8 wpi, and then collapsed with a significant reduction at 12 wpi. The nitrobenzene oxidation products of oil palm root lignin yielded both syringyl and guaiacyl monomers. Accumulation of lignin in the infected seedlings was in parallel to increased syringyl monomers, at 4 and 8 wpi. The activities of PAL and CAD enzymes in the infected seedlings at DS = 5–34% were significantly higher than the control seedlings and thereafter collapsed at DS = 69%.

  1. Agreement between activPAL and ActiGraph for assessing children's sedentary time

    Directory of Open Access Journals (Sweden)

    Ridgers Nicola D

    2012-02-01

    Full Text Available Abstract Background Accelerometers have been used to determine the amount of time that children spend sedentary. However, as time spent sitting may be detrimental to health, research is needed to examine whether accelerometer sedentary cut-points reflect the amount of time children spend sitting. The aim of this study was to: a examine agreement between ActiGraph (AG cut-points for sedentary time and objectively-assessed periods of free-living sitting and sitting plus standing time using the activPAL (aP; and b identify cut-points to determine time spent sitting and sitting plus standing. Methods Forty-eight children (54% boys aged 8-12 years wore a waist-mounted AG and thigh-mounted aP for two consecutive school days (9-3:30 pm. AG data were analyzed using 17 cut-points between 50-850 counts·min-1 in 50 counts·min-1 increments to determine sedentary time during class-time, break time and school hours. Sitting and sitting plus standing time were obtained from the aP for these periods. Limits of agreement were computed to evaluate bias between AG50 to AG850 sedentary time and sitting and sitting plus standing time. Receiver Operator Characteristic (ROC analyses identified AG cut-points that maximized sensitivity and specificity for sitting and sitting plus standing time. Results The smallest mean bias between aP sitting time and AG sedentary time was AG150 for class time (3.8 minutes, AG50 for break time (-0.8 minutes, and AG100 for school hours (-5.2 minutes. For sitting plus standing time, the smallest bias was observed for AG850. ROC analyses revealed an optimal cut-point of 96 counts·min-1 (AUC = 0.75 for sitting time, which had acceptable sensitivity (71.7% and specificity (67.8%. No optimal cut-point was obtained for sitting plus standing (AUC = 0.51. Conclusions Estimates of free-living sitting time in children during school hours can be obtained using an AG cut-point of 100 counts·min-1. Higher sedentary cut-points may capture both

  2. Refeeding syndrome in a young woman with argininosuccinate lyase deficiency

    Directory of Open Access Journals (Sweden)

    M. Stuy

    2015-09-01

    Full Text Available A severely chronically protein and calorie restricted young woman with argininosuccinate lyase deficiency developed transient refeeding syndrome (RFS and hyperammonemia after modest diet liberalization following initiation of glycerol phenylbutyrate (GPB. The patient required IV supportive care and supplementation with potassium, magnesium and calcium. She is now doing well on GPB and an appropriate maintenance diet. Susceptibility to RFS should be considered in chronically nutritionally restricted patients with metabolic disorders after liberalization of diet.

  3. L’opinion dissidente de Pal et le révisionnisme historique dans le Japon d’après-guerre The dissentient judgment of Pal and historical revisionism in Postwar Japan

    Directory of Open Access Journals (Sweden)

    Takeshi Nakajima

    2010-07-01

    Full Text Available Dans son opinion dissidente au procès de Tôkyô (le Tribunal militaire international pour l’Extrême-Orient, le juge indien Radhabinhod Pal avait contesté le bien-fondé des notions de « crime contre la paix » et de « crimes contre l’humanité » élaborées par la Charte du tribunal qui furent utilisées pour condamner les hauts responsables japonais comme « criminels de guerre de classe A ». Considérant que ces notions relèvent d’une législation ex post facto, il estima qu’il eût été préférable de s’en tenir à la notion existante de « crimes de guerre ». Faute de quoi, au lieu de servir la justice, le procès de Tokyo cautionna selon lui l’idée qu’il suffirait de sortir vainqueur d’un conflit armé pour pouvoir imposer la loi de manière arbitraire. Au Japon, cette opinion se vit peu à peu durcie, puis détournée de sa visée initiale, par un courant de plus en plus outrageusement révisionniste. Cet article réexamine les points essentiels de l’argumentation de Pal avant d’analyser la façon dont les révisionnistes japonais s’en sont emparée.In his dissenting judgment in the Tokyo trial (the International Military Tribunal for the Far East, the Indian judge Radhabinhod Pal had challenged the validity of the concepts of "crime against peace" and "crime against humanity" developed by the Charter of the court and which were used to condemn the Japanese officials as "war criminals category A". Whereas these concepts are subject to ex post facto legislation, he felt it was better to stick to the existing concept of "war crimes". Failing that, instead of justice, the trial of Tokyo according to endorse the idea that it was enough to win an armed conflict to impose the law in an arbitrary manner. In Japan, this opinion is gradually tightened, then diverted from its original purpose, by a stream of more and more outrageously revisionist. This article reviews the essential points of the argument of

  4. Regulation of expression of pectate lyase genes pelA, pelD, and pelE in Erwinia chrysanthemi.

    Science.gov (United States)

    Reverchon, S; Robert-Baudouy, J

    1987-06-01

    The regulation of pelA, pelD, and pelE genes encoding three of the five major pectate lyase isoenzymes (PLa, PLd, and PLe) in Erwinia chrysanthemi B374 was analyzed by using genetic fusions to lacZ. These three genes are clustered on a 5-kilobase DNA fragment in the order pelD-pelE-pelA and constitute three independent transcriptional units. We localized the pelDEA cluster near the pro-1 marker on the genetic map of B374 by chromosomal mobilization with RP4::mini-Mu plasmid pULB110. Three classes of regulatory mutations responsible for constitutive pectate lyase synthesis have been described (kdgR, gpiR, and cri). We studied the effects of each mutation on pelE, pelD, and pelA expression independently. The mutations kdgR and gpiR mainly affect the expression of pelE and pelD, although PLa synthesis is slightly increased. The cri mutation results in a low level of constitutive expression of the three pel genes, but it is a pleiotropic mutation since other genes not involved in pectinolysis are also affected. In addition, we demonstrated that exuR, a negative regulatory gene governing the catabolism of hexuronates, does not modify the expression of pel genes. The frequency of gpiR or cri mutations (about 10(-8)) and the resulting constitutivity of pectate lyase synthesis suggest that these genes act as negative regulatory genes in addition to kdgR, which is already known to encode a repressor. Moreover, we found that expression of pel-lac fusions carried on pBR322 derivatives was higher in E. chrysanthemi than in Escherichia coli; this fact suggests the existence of positive regulation of pectate lyase synthesis in E. chrysanthemi.

  5. Molecular cloning and characterization of an Erwinia carotovora subsp. carotovora pectin lyase gene that responds to DNA-damaging agents.

    OpenAIRE

    McEvoy, J L; Murata, H; Chatterjee, A K

    1990-01-01

    recA-mediated production of pectin lyase (PNL) and the bacteriocin carotovoricin occurs in Erwinia carotovora subsp. carotovora 71 when this organism is subjected to agents that damage or inhibit the synthesis of DNA. The structural gene pnlA was isolated from a strain 71 cosmid gene library following mobilization of the cosmids into a moderate PNL producer, strain 193. The cosmid complemented pnl::Tn5 but not ctv::Tn5 mutations. A constitutive level of PNL activity was detected in RecA+ and ...

  6. Biochemical study of leaf browning in minimally processed leaves of lettuce (Lactuca sativa L. var. acephala).

    Science.gov (United States)

    Degl'Innocenti, E; Guidi, L; Pardossi, A; Tognoni, F

    2005-12-28

    A series of biochemical parameters, including the concentration of total ascorbic acid (ASA(tot)) and the activities of phenylalanine ammonia lyase (PAL), polyphenol oxidase (PPO), and peroxidases (PODs), was investigated during cold storage (72 h at 4 degrees C in the dark) in fresh-cut (minimally processed) leaves of two lettuce (Lactuca sativa L. var. acephala) cultivars differing in the susceptibility to tissue browning: Green Salade Bowl (GSB), susceptible, and Red Salade Bowl (RSB), resistant. The two cultivars showed differences also at the biochemical level. The content in ASA(tot) increased in RSB, as a consequence of increased DHA concentration; conversely, ASA(tot) diminished in GSB, in which ASA was not detectable after 72 h of storage, thus suggesting a disappearance of ascorbate (both ASA and DHA) into nonactive forms. The antioxidant capacity (as determined by using FRAP analysis) decreased significantly during storage in RSB, while a strong increase was observed in GSB. PAL activity increased soon after processing reaching a maximum by 3 h, then it declined to a relatively constant value in RSB, while in GSB it showed a tendency to decrease in the first few hours from harvest and processing. POD activity, at least for chlorogenic acid, increased significantly during storage only in GSB.

  7. Oxidative and Molecular Responses in Capsicum annuum L. after Hydrogen Peroxide, Salicylic Acid and Chitosan Foliar Applications

    Science.gov (United States)

    Mejía-Teniente, Laura; de Dalia Durán-Flores, Flor; Chapa-Oliver, Angela María; Torres-Pacheco, Irineo; Cruz-Hernández, Andrés; González-Chavira, Mario M.; Ocampo-Velázquez, Rosalía V.; Guevara-González, Ramón G.

    2013-01-01

    Hydrogen peroxide (H2O2) is an important ROS molecule (Reactive oxygen species) that serves as a signal of oxidative stress and activation of signaling cascades as a result of the early response of the plant to biotic stress. This response can also be generated with the application of elicitors, stable molecules that induce the activation of transduction cascades and hormonal pathways, which trigger induced resistance to environmental stress. In this work, we evaluated the endogenous H2O2 production caused by salicylic acid (SA), chitosan (QN), and H2O2 elicitors in Capsicum annuum L. Hydrogen peroxide production after elicitation, catalase (CAT) and phenylalanine ammonia lyase (PAL) activities, as well as gene expression analysis of cat1, pal, and pathogenesis-related protein 1 (pr1) were determined. Our results displayed that 6.7 and 10 mM SA concentrations, and, 14 and 18 mM H2O2 concentrations, induced an endogenous H2O2 and gene expression. QN treatments induced the same responses in lesser proportion than the other two elicitors. Endogenous H2O2 production monitored during several days, showed results that could be an indicator for determining application opportunity uses in agriculture for maintaining plant alert systems against a stress. PMID:23676352

  8. Structural insights into RipC, a putative citrate lyase β subunit from a Yersinia pestis virulence operon

    International Nuclear Information System (INIS)

    Torres, Rodrigo; Chim, Nicholas; Sankaran, Banumathi; Pujol, Céline; Bliska, James B.; Goulding, Celia W.

    2011-01-01

    Comparison of the 2.45 Å resolution crystal structure of homotrimeric RipC, a putative citrate lyase β subunit from Y. pestis, with structural homologs reveals conserved RipC residues that are implicated in CoA binding. Yersinia pestis remains a threat, with outbreaks of plague occurring in rural areas and its emergence as a weapon of bioterrorism; thus, an improved understanding of its various pathogenicity pathways is warranted. The rip (required for intracellular proliferation) virulence operon is required for Y. pestis survival in interferon-γ-treated macrophages and has been implicated in lowering macrophage-produced nitric oxide levels. RipC, one of three gene products from the rip operon, is annotated as a citrate lyase β subunit. Furthermore, the Y. pestis genome lacks genes that encode citrate lyase α and γ subunits, suggesting a unique functional role of RipC in the Y. pestisrip-mediated survival pathway. Here, the 2.45 Å resolution crystal structure of RipC revealed a homotrimer in which each monomer consists of a (β/α) 8 TIM-barrel fold. Furthermore, the trimeric state was confirmed in solution by size-exclusion chromatography. Through sequence and structure comparisons with homologous proteins, it is proposed that RipC is a putative CoA- or CoA-derivative binding protein

  9. [Assessment of IDC-Pal as a Diagnostic Tool for Family Physicians to Identify Patients with Complex Palliative Care Needs in Germany: a Pilot Study].

    Science.gov (United States)

    Comino, María Rosa Salvador; Garcia, Victor Regife; López, Maria Auxiliadora Fernández; Feddersen, Berend; Roselló, María Luisa Martin; Sanftenberg, Linda; Schelling, Jörg

    2017-07-11

    Background Palliative medicine is an essential component of the health care system. Basic palliative care should be provided by primary care services (family physician and home nursing) with palliative-medical basic qualification. Often it is very difficult to identify patients that would profit from a specialized palliative care team. For the evaluation of the case complexity of a palliative patient, we present a Spanish diagnostic tool IDC-Pal, which tries to specify when, why and where a palliative patient should be referred. The aims of this study were the translation and cultural adaptation of the diagnostic tool for complexity in palliative care IDC-Pal to the German language, and the measurement of its feasibility and face validity. Methods During the first phase, a forward-backward translation with linguistic and cultural adaptation of the tool IDC-Pal as well as the validation of its content by a review committee was performed. During the second phase, the preliminary version of the tool was tested by 38 family physicians that were asked for a qualitative assessment using a 10-points Likert scale (1 = "strongly disagree" and 10 = "totally agree"). Finally, a definitive version was developed. Results The translation and adaptation were achieved without major problems. Both feasibility and apparent validity of the tool IDC-Pal were rated as high. The mean response in the Likert scale was 7.79, with a SD of 0.36. Participants strongly agreed on the apparent validity of the tool with a mean of 7.82 and a SD of 0.26 and on its feasibility with a mean of 7.79, and a SD of 0.39. Conclusions A conceptually, culturally and linguistically equivalent version of the original instrument IDC-Pal was obtained. German family physicians agreed on the usability of IDC-Pal as a tool for rating the case complexity of palliative patients. These results indicate that physicians in Bavaria and eventually in Germany could benefit of the full validation of IDC-Pal. © Georg Thieme

  10. Structural insights into the bacterial carbon - phosphorus lyase machinery

    DEFF Research Database (Denmark)

    Seweryn, Paulina; Van, Lan Bich; Kjeldgaard, Morten

    2015-01-01

    Phosphorus is required for all life and microorganisms can extract it from their environment through several metabolic pathways. When phosphate is in limited supply, some bacteria are able to use phosphonate compounds, which require specialized enzymatic machinery to break the stable carbon......–phosphorus (C–P) bond. Despite its importance, the details of how this machinery catabolizes phosphonates remain unknown. Here we determine the crystal structure of the 240-kilodalton Escherichia coli C–P lyase core complex (PhnG–PhnH–PhnI–PhnJ; PhnGHIJ), and show that it is a two-fold symmetric hetero...

  11. Step detection and activity recognition accuracy of seven physical activity monitors.

    Directory of Open Access Journals (Sweden)

    Fabio A Storm

    Full Text Available The aim of this study was to compare the seven following commercially available activity monitors in terms of step count detection accuracy: Movemonitor (Mc Roberts, Up (Jawbone, One (Fitbit, ActivPAL (PAL Technologies Ltd., Nike+ Fuelband (Nike Inc., Tractivity (Kineteks Corp. and Sensewear Armband Mini (Bodymedia. Sixteen healthy adults consented to take part in the study. The experimental protocol included walking along an indoor straight walkway, descending and ascending 24 steps, free outdoor walking and free indoor walking. These tasks were repeated at three self-selected walking speeds. Angular velocity signals collected at both shanks using two wireless inertial measurement units (OPAL, ADPM Inc were used as a reference for the step count, computed using previously validated algorithms. Step detection accuracy was assessed using the mean absolute percentage error computed for each sensor. The Movemonitor and the ActivPAL were also tested within a nine-minute activity recognition protocol, during which the participants performed a set of complex tasks. Posture classifications were obtained from the two monitors and expressed as a percentage of the total task duration. The Movemonitor, One, ActivPAL, Nike+ Fuelband and Sensewear Armband Mini underestimated the number of steps in all the observed walking speeds, whereas the Tractivity significantly overestimated step count. The Movemonitor was the best performing sensor, with an error lower than 2% at all speeds and the smallest error obtained in the outdoor walking. The activity recognition protocol showed that the Movemonitor performed best in the walking recognition, but had difficulty in discriminating between standing and sitting. Results of this study can be used to inform choice of a monitor for specific applications.

  12. Step detection and activity recognition accuracy of seven physical activity monitors.

    Science.gov (United States)

    Storm, Fabio A; Heller, Ben W; Mazzà, Claudia

    2015-01-01

    The aim of this study was to compare the seven following commercially available activity monitors in terms of step count detection accuracy: Movemonitor (Mc Roberts), Up (Jawbone), One (Fitbit), ActivPAL (PAL Technologies Ltd.), Nike+ Fuelband (Nike Inc.), Tractivity (Kineteks Corp.) and Sensewear Armband Mini (Bodymedia). Sixteen healthy adults consented to take part in the study. The experimental protocol included walking along an indoor straight walkway, descending and ascending 24 steps, free outdoor walking and free indoor walking. These tasks were repeated at three self-selected walking speeds. Angular velocity signals collected at both shanks using two wireless inertial measurement units (OPAL, ADPM Inc) were used as a reference for the step count, computed using previously validated algorithms. Step detection accuracy was assessed using the mean absolute percentage error computed for each sensor. The Movemonitor and the ActivPAL were also tested within a nine-minute activity recognition protocol, during which the participants performed a set of complex tasks. Posture classifications were obtained from the two monitors and expressed as a percentage of the total task duration. The Movemonitor, One, ActivPAL, Nike+ Fuelband and Sensewear Armband Mini underestimated the number of steps in all the observed walking speeds, whereas the Tractivity significantly overestimated step count. The Movemonitor was the best performing sensor, with an error lower than 2% at all speeds and the smallest error obtained in the outdoor walking. The activity recognition protocol showed that the Movemonitor performed best in the walking recognition, but had difficulty in discriminating between standing and sitting. Results of this study can be used to inform choice of a monitor for specific applications.

  13. PALS investigations of matrix Vycor glass doped with molecules of luminescent dye and silver nanoparticles. Discrepancies from the ETE model

    Directory of Open Access Journals (Sweden)

    Gorgol Marek

    2015-12-01

    Full Text Available A thermal stability of three materials: undoped reference Vycor glass, glass filled with ROT-305 red dye, and silver nanoparticles was investigated by positron annihilation lifetime spectroscopy (PALS in a broad temperature range (from 93 to 473 K. The attempt of pore size calculations from the ortho-positronium lifetime data was performed using the extended Tao-Eldrup (ETE model. Below room temperature, a significant decrease in lifetime values of the longest-lived component was found for all the samples. This effect could not be explained by thermal shrinkage of the material and is probably caused by interaction of o-Ps with a Vycor glass matrix. The greatest discrepancy from the ETE model predictions was observed for the reference glass. Doping the base material with dye molecules and silver nanoparticles resulted in similar small decrease in this discrepancy. After reheating the samples to the room temperature, the PALS components returned to the initial values. In the temperature range of 293–473 K, quite good agreement between PALS results and the ETE model predictions was observed for the reference glass and the glass incorporated with dye molecules. The observed small discrepancy in this range could possibly be partly explained by thermal expansion of the material. For the glass doped with silver nanoparticles, a significant change in PALS parameters was observed in the temperature range from 403 to 473 K.

  14. Investigations of mechanism of laser radiation absorption at PALS

    Czech Academy of Sciences Publication Activity Database

    Kalinowska, Z.; Kasperczuk, A.; Pisarczyk, T.; Chodukowski, T.; Gus’kov, S.Yu.; Demchenko, N. N.; Ullschmied, Jiří; Krouský, Eduard; Pfeifer, Miroslav; Skála, Jiří

    2012-01-01

    Roč. 57, č. 2 (2012), s. 227-230 ISSN 0029-5922. [International Conference on Research and Applications of Plasmas (PLASMA)2011. Warsaw, 12.09.2011-16.09.2011] R&D Projects: GA MŠk(CZ) LC528 Institutional research plan: CEZ:AV0Z20430508 Keywords : collisional absorption * crater volume * electron density distribution * interferometry * iodine laser * resonance absorption * Laser radiation * inverse bremsstrahlung * laser interferometry * PALS laser Subject RIV: BL - Plasma and Gas Discharge Physics Impact factor: 0.507, year: 2012 http://www.nukleonika.pl/www/back/full/vol57_2012/v57n2p227f.pdf

  15. 1sn06-56 Dogbo.p65

    African Journals Online (AJOL)

    Sci-Nat

    résistantes aux microorganismes. ... la lumière naturelle. La photopériode au cours de l'expérimentation a varié entre 12 h et 13 h (mai, juin, juillet 2005) et la température entre 28 °C et. 32 °C. La ..... Tableau 1: Effets de l'acide salicylique sur l'activité de la phénylalanine ammonia-lyase (PAL) dans les feuilles de plants de ...

  16. Crystal structure and mechanism of the Staphylococcus cohnii virginiamycin B lyase (Vgb).

    Science.gov (United States)

    Lipka, Magdalena; Filipek, Renata; Bochtler, Matthias

    2008-04-08

    The semisynthetic streptogramin antibiotic quinupristin/dalfopristin (trade name Synercid, Aventis Pharma) is a mixture of the A-type streptogramin dalfopristin and the B-type streptogramin quinupristin, a capped hexapeptide macrolactone. Quinupristin/dalfopristin was developed to combat multidrug resistant pathogens, but suffers from its own problems with drug resistance. Virginiamycin B lyase (Vgb) inactivates the quinupristin component of Synercid by lactone ring opening. Remarkably, the enzyme promotes this reaction by intramolecular beta-elimination without the involvement of a water molecule. Recently, structures of S. aureus Vgb in the presence and absence of substrate were reported and used together with detailed mutagenesis data to suggest a catalytic mechanism. Here, we report an independent determination of the S. cohnii Vgb crystal structure and a biochemical characterization of the enzyme. As expected, the S. cohnii and S. aureus Vgb structures and active sites are very similar. Moreover, both enzymes catalyze quinupristin lactone ring opening with similar rate constants, albeit perhaps with different dependencies on divalent metal ions. Replacement of the conserved active site residues His228, Glu268, or His270 with alanine reduces or abolishes S. cohnii Vgb activity. Residue Lys285 in S. cohnii Vgb is spatially equivalent to the S. aureus Vgb active site residue Glu284. A glutamate but not an alanine residue can substitute for the lysine without significant loss of activity.

  17. A horizontal gene transfer at the origin of phenylpropanoid metabolism: a key adaptation of plants to land

    Directory of Open Access Journals (Sweden)

    Gribaldo Simonetta

    2009-02-01

    Full Text Available Abstract Background The pioneering ancestor of land plants that conquered terrestrial habitats around 500 million years ago had to face dramatic stresses including UV radiation, desiccation, and microbial attack. This drove a number of adaptations, among which the emergence of the phenylpropanoid pathway was crucial, leading to essential compounds such as flavonoids and lignin. However, the origin of this specific land plant secondary metabolism has not been clarified. Results We have performed an extensive analysis of the taxonomic distribution and phylogeny of Phenylalanine Ammonia Lyase (PAL, which catalyses the first and essential step of the general phenylpropanoid pathway, leading from phenylalanine to p-Coumaric acid and p-Coumaroyl-CoA, the entry points of the flavonoids and lignin routes. We obtained robust evidence that the ancestor of land plants acquired a PAL via horizontal gene transfer (HGT during symbioses with soil bacteria and fungi that are known to have established very early during the first steps of land colonization. This horizontally acquired PAL represented then the basis for further development of the phenylpropanoid pathway and plant radiation on terrestrial environments. Conclusion Our results highlight a possible crucial role of HGT from soil bacteria in the path leading to land colonization by plants and their subsequent evolution. The few functional characterizations of sediment/soil bacterial PAL (production of secondary metabolites with powerful antimicrobial activity or production of pigments suggest that the initial advantage of this horizontally acquired PAL in the ancestor of land plants might have been either defense against an already developed microbial community and/or protection against UV. Reviewers This article was reviewed by Purificación López-García, Janet Siefert, and Eugene Koonin.

  18. Des villages au Népal se préparent à des conditions climatiques ...

    International Development Research Centre (IDRC) Digital Library (Canada)

    4 avr. 2012 ... La grande diversité ethnique, culturelle et linguistique, ainsi que l'inaccessibilité de nombreuses communautés rurales, particulièrement dans les régions montagneuses, font partie des facteurs venant compliquer les choses. La recherche. En 2010, l'Institute for Social and Environmental Transition – Népal ...

  19. Rhodotorulaglutinis phenylalanine/tyrosine ammonia lyase enzyme catalyzed synthesis of the methyl ester of para-hydroxycinnamic acid and its potential antibacterial activity

    Directory of Open Access Journals (Sweden)

    Marybeth C MacDonald

    2016-03-01

    Full Text Available Biotransformation of L-tyrosine methyl ester (L-TM to the methyl ester of para- hydroxycinnamic acid (p-HCAM using Rhodotorula glutinis yeast phenylalanine/tyrosine ammonia lyase (PTAL; EC 4.3.1.26 enzyme was successfully demonstrated for the first time; progress of the reaction was followed by spectrophotometric determination at 315 nm. The following conditions were optimized for maximal formation of p-HCAM: pH (8.5, temperature (37 C, speed of agitation (50 rpm, enzyme concentration (0.080 µM, and substrate concentration (0.50 mM. Under these conditions, the yield of the reaction was ~15% in 1 h incubation period and ~63% after an overnight (~18 h incubation period. The product (p-HCAM of the reaction of PTAL with L-TM was confirmed using Nuclear Magnetic Resonance spectroscopy (NMR. Fourier Transform Infra-Red spectroscopy (FTIR was carried out to rule out potential hydrolysis of p-HCAM during overnight incubation. Potential antibacterial activity of p-HCAM was tested against several strains of Gram positive and Gram negative bacteria. This study describes a synthetically useful transformation, and could have future clinical and industrial applications.

  20. Analysis and prediction of daily physical activity level data using autoregressive integrated moving average models

    NARCIS (Netherlands)

    Long, Xi; Pauws, S.C.; Pijl, M.; Lacroix, J.; Goris, A.H.C.; Aarts, R.M.

    2009-01-01

    Results are provided on predicting daily physical activity level (PAL) data from past data of participants of a physical activity lifestyle program aimed at promoting a healthier lifestyle consisting of more physical exercise. The PAL data quantifies the level of a person’s daily physical activity

  1. Cell differentiation during sexual development of the fungus Sordaria macrospora requires ATP citrate lyase activity.

    Science.gov (United States)

    Nowrousian, M; Masloff, S; Pöggeler, S; Kück, U

    1999-01-01

    During sexual development, mycelial cells from most filamentous fungi differentiate into typical fruiting bodies. Here, we describe the isolation and characterization of the Sordaria macrospora developmental mutant per5, which exhibits a sterile phenotype with defects in fruiting body maturation. Cytological investigations revealed that the mutant strain forms only ascus precursors without any mature spores. Using an indexed cosmid library, we were able to complement the mutant to fertility by DNA-mediated transformation. A single cosmid clone, carrying a 3.5-kb region able to complement the mutant phenotype, has been identified. Sequencing of the 3.5-kb region revealed an open reading frame of 2.1 kb interrupted by a 66-bp intron. The predicted polypeptide (674 amino acids) shows significant homology to eukaryotic ATP citrate lyases (ACLs), with 62 to 65% amino acid identity, and the gene was named acl1. The molecular mass of the S. macrospora ACL1 polypeptide is 73 kDa, as was verified by Western blot analysis with a hemagglutinin (HA) epitope-tagged ACL1 polypeptide. Immunological in situ detection of the HA-tagged polypeptide demonstrated that ACL is located within the cytosol. Sequencing of the mutant acl1 gene revealed a 1-nucleotide transition within the coding region, resulting in an amino acid substitution within the predicted polypeptide. Further evidence that ACL1 is essential for fruiting body maturation comes from experiments in which truncated and mutated versions of the acl1 gene were used for transformation. None of these copies was able to reconstitute the fertile phenotype in transformed per5 recipient strains. ACLs are usually involved in the formation of cytosolic acetyl coenzyme A (acetyl-CoA), which is used for the biosynthesis of fatty acids and sterols. Protein extracts from the mutant strain showed a drastic reduction in enzymatic activity compared to values obtained from the wild-type strain. Investigation of the time course of ACL

  2. Characterization of a new (R)-hydroxynitrile lyase from the Japanese apricot Prunus mume and cDNA cloning and secretory expression of one of the isozymes in Pichia pastoris.

    Science.gov (United States)

    Fukuta, Yasuhisa; Nanda, Samik; Kato, Yasuo; Yurimoto, Hiroya; Sakai, Yasuyoshi; Komeda, Hidenobu; Asano, Yasuhisa

    2011-01-01

    PmHNL, a hydroxynitrile lyase from Japanese apricot ume (Prunus mume) seed was purified to homogeneity by ammonium sulfate fractionation and chromatographic steps. The purified enzyme was a monomer with molecular mass of 58 kDa. It was a flavoprotein similar to other hydroxynitrile lyases of the Rosaceae family. It was active over a broad temperature, and pH range. The N-terminal amino acid sequence (20 amino acids) was identical with that of the enzyme from almond (Prunus dulcis). Based on the N-terminal sequence of the purified enzyme and the conserved amino acid sequences of the enzymes from Pr. dulcis, inverse PCR method was used for cloning of a putative PmHNL (PmHNL2) gene from a Pr. mume seedling. Then the cDNA for the enzyme was cloned. The deduced amino acid sequence was found to be highly similar (95%) to that of an enzyme from Pr. serotina, isozyme 2. The recombinant Pichia pastoris transformed with the PmHNL2 gene secreted an active enzyme in glycosylated form.

  3. One Year Experience of Pheburane® (Sodium Phenylbutyrate) Treatment in a Patient with Argininosuccinate Lyase Deficiency

    OpenAIRE

    Uçar, Sema Kalkan; Ozbaran, Burcu; Altinok, Yasemin Atik; Kose, Melis; Canda, Ebru; Kagnici, Mehtap; Coker, Mahmut

    2015-01-01

    Argininosuccinate lyase deficiency (ASLD) is a urea cycle disorder (UCD) treated with dietary adjustment and nitrogen scavenging agents. “Pheburane®” is a new tasteless and odour-free formulation of sodium phenylbutyrate, indicated in the treatment of UCD.

  4. Alterations in antioxidant metabolism in coffee leaves infected by Cercospora coffeicola

    Directory of Open Access Journals (Sweden)

    Camila Cristina Lage de Andrade

    2016-01-01

    Full Text Available ABSTRACT: Brown eye spot (BE caused by Cercospora coffeicola is the main disease of coffee crop. A variation in symptoms of BE has been reported in the field, raising suspicion of occurrence of new species. However, information about coffee- C. coffeicola interaction is still limited. This research aimed to determine the difference between antioxidant metabolism of coffee plants cultivar Mundo Novo inoculated with a strain isolated from a common BE lesion (CML 2984 and a strain isolated from a black BE lesion (CML 2985. The enzyme activity of peroxidase (POX, catalase (CAT, superoxide dismutase (SOD, ascorbate peroxidase (APX and phenylalanine ammonia lyase (PAL were determined. Activities of POX, APX, and PAL increased in plants inoculated with both strains compared to non-inoculated plants at 12 and 24 hours post inoculation (hpi. CAT activity increased in inoculated plants with black BE strain at 24 hpi and both strains at 48 hpi. The SOD activity only increased in inoculated plants with both strains at 48 hpi. These results show that an elevated antioxidant response was observed when the plants were challenged with both strains of C. coffeicola. Both strains produced lesions of the common type, suggesting that other factors lead to the development of black BE lesion type under field conditions and further investigation is needed.

  5. Increased expression of endothelial antigen PAL-E in human diabetic retinopathy correlates with microvascular leakage

    NARCIS (Netherlands)

    Schlingemann, R. O.; Hofman, P.; Vrensen, G. F.; Blaauwgeers, H. G.

    1999-01-01

    AIMS/HYPOTHESIS: The Pathologische Anatomie Leiden-Endothelium (PAL-E) antigen is a marker for loss of the blood-brain barrier function in brain tumours. It is endothelium specific and is associated with the endothelial plasmalemmal vesicles (caveolae) involved in transcellular transport. To test

  6. Correlates of physical activity level among hemodialysis patients in Selangor, Malaysia.

    Science.gov (United States)

    Wong, S W; Chan, Y M; Lim, T S

    2011-12-01

    There is mounting evidence demonstrating the importance of adequate physical activity to promote better well-being among hemodialysis patients. Available data pertaining to the levels of physical activity and its determinants among hemodialysis patients is, however, scarce in Malaysia. The objectives of this study are hence to determine the levels of physical activity and it associated factors among hemodialysis patients. A total of 70 subjects were recruited from three dialysis centres in Selangor. A face-to-face interview was conducted to obtain socio-demographic data and subjects' knowledge on dietary sources. Medical history, biochemical parameters and weight status were obtained from medical records. Physical activity level (PAL) was assessed using the Global Physical Activity Questionnaire (GPAQ). A total of 81.4% and 18.6% of the respondents had low and moderate PALs, respectively. Thus, none of the respondents had high PAL. Serum creatinine, education level, personal income and knowledge score on potassium-related medical complications were factors found to correlate significantly with PAL. Multiple linear regression analysis showed that higher PAL was predicted by a lower knowledge score on dietary sodium source, higher education and higher serum creatinine. Despite consistent documentation of the potential positive impact of physical exercise on hemodialysis outcomes, the level of physical activity remains low among these patients. It is hoped that these findings can add to the existing body of knowledge and serve as a supporting document for the formulation of appropriate interventions to improve the status of physical activity among hemodialysis patients in Malaysia.

  7. Salicylic Acid Alleviates Aluminum Toxicity in Soybean Roots through Modulation of Reactive Oxygen Species Metabolism

    Directory of Open Access Journals (Sweden)

    Ning Liu

    2017-11-01

    Full Text Available As an important signal molecule, salicylic acid (SA improves plant tolerance to aluminum (Al stress. The objective of this study was to investigate the effects of exogenous SA application on the dynamics of endogenous SA and reactive oxygen species in soybean (Glycine max L. exposed to Al stress. The roots of soybean seedlings were exposed to a combination of AlCl3 (30 μM and SA (10 μM/PAC (100 μM, paclobutrazol, SA biosynthesis inhibitor for 3, 6, 9, and 12 h. Al stress induced an increase in endogenous SA concentration in a time-dependent manner, also verified by the up-regulated expression of GmNPR1, an SA-responsive gene. Al stress increased the activities of phenylalanine ammonia-lyase (PAL and benzoic acid 2-hydroxylase (BA2H, and the contents of SA, O2- and malondialdehyde (MDA in the root apex. The application of exogenous SA increased PAL and BA2H, and reduced O2- and MDA contents in soybean roots under Al stress. PAC inhibited the SA induced increase in BA2H activity. In addition, the SA application resulted in a rapid increase in hydrogen peroxide (H2O2 concentration under Al stress, followed by a sharp decrease. Compared with the plants exposed to Al alone, Al+SA plants possessed higher activities of superoxide dismutase, peroxidase, and ascorbate peroxidase, and lower catalase activity, indicating that SA alleviated Al-induced oxidative damage. These results suggested that PAL and BA2H were involved in Al-induced SA production and showed that SA alleviated the adverse effects of Al toxicity by modulating the cellular H2O2 level and the antioxidant enzyme activities in the soybean root apex.

  8. Salicylic acid alleviates aluminum toxicity in soybean roots through modulation of reactive oxygen species metabolism

    Science.gov (United States)

    Liu, Ning; Song, Fengbin; Zhu, Xiancan; You, Jiangfeng; Yang, Zhenming; Li, Xiangnan

    2017-11-01

    As an important signal molecule, salicylic acid (SA) improves plant tolerance to aluminum (Al) stress. The objective of this study was to investigate the effects of exogenous SA application on the dynamics of endogenous SA and reactive oxygen species in soybean (Glycine max L.) exposed to Al stress. The roots of soybean seedlings were exposed to a combination of AlCl3 (30 μM) and SA (10 μM)/PAC (100 μM, paclobutrazol, SA biosynthesis inhibitor) for 3, 6, 9 and 12 h. Al stress induced an increase in endogenous SA concentration in a time-dependent manner, also verified by the up-regulated expression of GmNPR1, an SA-responsive gene. Al stress increased the activities of phenylalanine ammonia-lyase (PAL) and benzoic acid 2-hydroxylase (BA2H), and the contents of SA, O2- and malondialdehyde (MDA) in the root apex. The application of exogenous SA increased PAL and BA2H, and reduced O2- and MDA contents in soybean roots under Al stress. PAC inhibited the SA induced increase in BA2H activity. In addition, the SA application resulted in a rapid increase in hydrogen peroxide (H2O2) concentration under Al stress, followed by a sharp decrease. Compared with the plants exposed to Al alone, Al+SA plants possessed higher activities of superoxide dismutase, peroxidase and ascorbate peroxidase, and lower catalase activity, indicating that SA alleviated Al-induced oxidative damage. These results suggested that PAL and BA2H were involved in Al-induced SA production and showed that SA alleviated the adverse effects of Al toxicity by modulating the cellular H2O2 level and the antioxidant enzyme activities in the soybean root apex.

  9. Effects of sodium nitroprusside (SNP) pretreatment on UV-B stress tolerance in lettuce (Lactuca sativa L.) seedlings.

    Science.gov (United States)

    Esringu, Aslıhan; Aksakal, Ozkan; Tabay, Dilruba; Kara, Ayse Aydan

    2016-01-01

    Ultraviolet-B (UV-B) radiation is one of the most important abiotic stress factors that could influence plant growth, development, and productivity. Nitric oxide (NO) is an important plant growth regulator involved in a wide variety of physiological processes. In the present study, the possibility of enhancing UV-B stress tolerance of lettuce seedlings by the exogenous application of sodium nitroprusside (SNP) was investigated. UV-B radiation increased the activities of superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), peroxidase (POD) and total phenolic concentrations, antioxidant capacity, and expression of phenylalanine ammonia lyase (PAL) gene in seedlings, but the combination of SNP pretreatment and UV-B enhanced antioxidant enzyme activities, total phenolic concentrations, antioxidant capacity, and PAL gene expression even more. Moreover, UV-B radiation significantly inhibited chlorophylls, carotenoid, gibberellic acid (GA), and indole-3-acetic acid (IAA) contents and increased the contents of abscisic acid (ABA), salicylic acid (SA), malondialdehyde (MDA), hydrogen peroxide (H2O2), and superoxide radical (O2•(-)) in lettuce seedlings. When SNP pretreatment was combined with the UV-B radiation, we observed alleviated chlorophylls, carotenoid, GA, and IAA inhibition and decreased content of ABA, SA, MDA, H2O2, and O2•(-) in comparison to non-pretreated stressed seedlings.

  10. Are physical activity levels linked to nutrient adequacy? Implications for cancer risk.

    Science.gov (United States)

    Csizmadi, Ilona; Kelemen, Linda E; Speidel, Thomas; Yuan, Yan; Dale, Laura C; Friedenreich, Christine M; Robson, Paula J

    2014-01-01

    Cancer prevention guidelines recommend a healthy body mass index, physical activity, and nutrient intake from food rather than supplements. Sedentary individuals may restrict energy intake to prevent weight gain and in so doing may compromise nutritional intake. We conducted a cross-sectional analysis to determine if adequacy of micronutrients is linked to physical activity levels (PALs) in healthy-weight adults. Tomorrow Project participants in Alberta, Canada (n = 5333), completed past-year diet and physical activity questionnaires. The percent meeting Dietary Reference Intakes (DRIs) was reported across low and high PAL groups, and the relation between PAL and percent achieved DRI was determined using multiple linear regression analyses. Overall, genders (P physical activity to include having a more favorable impact on nutrient adequacy.

  11. In situ localization of phenylpropanoid biosynthetic mRNAs and proteins in Parsley (Petroselinum crispum)

    International Nuclear Information System (INIS)

    Reinold, S.; Hahlbrock, K.

    1997-01-01

    Using in situ RNA/RNA hybridization, enzyme immunolocalization, and histochemical techniques, several phenylpropanoid biosynthetic activities and products were localized in tissue sections from various aerial parts of parsley (Petroselinum crispum) plants at different developmental stages. The enzymes and corresponding mRNAs analyzed included two representatives of general phenylpropanoid metabolism: phenylalanine ammonia-lyase (PAL) and 4-coumarate: CoA ligase (4CL), and one representative each from two distinct branch pathways: chalcone synthase (CHS; flavonoids) and S-adenosyl-L-methionine: bergaptol O-methyltransferase (BMT; furanocoumarins). In almost all cases, the relative timing of accumulation differed greatly for mRNA and protein and indicated short expression periods and short half-lives for all mRNAs as compared to the proteins. PAL and 4CL occurred almost ubiquitously in cell type-specific patterns, and their mRNAs and proteins were always coordinately expressed, whereas the cell type-specific localization of flavonoid and furanocoumarin biosynthetic activities was to a large extent mutually exclusive. However, the distribution patterns of CHS and BMT, when superimposed, closely matched those of PAL and 4CL in nearly all tissues analysed, suggesting that the flavonoid and furanocoumarin pathways together constituted a large majority of the total phenylpropanoid biosynthetic activity. Differential sites of synthesis and accumulation indicating intercellular translocation were observed both for flavonoids and for furanocoumarins in oil ducts and the surrounding tissue. The widespread occurrence of both classes of compounds, as well as selected, pathway-specific mRNAs and enzymes, in many cell types of all parsley organs including various flower parts suggests additional functions beyond the previously established roles of flavonoids in UV protection and furanocoumarins in pathogen defence. (author)

  12. Structural analysis of poly(vinyl alcohol hydrogels) produced by freezing and thawing process Positron annihilation lifetime spectroscopy (PALS)

    International Nuclear Information System (INIS)

    Paranhos, Caio M.; Oliveira, Renata N.; Soares, Bluma G.

    2005-01-01

    In this work, poly(vinyl alcohol) hydrogels with a entrapped polyelectrolyte were characterized through Positron Annihilation Lifetime Spectroscopy (PALS). PALS was used to determine the average pore radii (R), average free volume (V f ) and the intensity of ortho-positronium (I 3 ). The free volume estimated for the hydrogels increases with increasing polyelectrolyte content, while the intensity of o-Ps decreases with the increasing polyelectrolyte content. Degree of crystallinity of the samples decreases with the presence of polyelectrolyte. The structural changes observed can be discussed in terms of the changes in hydrogels structure due to a hindrance capacity of the polyelectrolyte on the crystalline behavior and the different polymer-polymer interactions. (author)

  13. Cystathionine γ-lyase deficiency mediates neurodegeneration in Huntington’s disease

    Science.gov (United States)

    Paul, Bindu D.; Sbodio, Juan I.; Xu, Risheng; Vandiver, M. Scott; Cha, Jiyoung Y.; Snowman, Adele M.; Snyder, Solomon H.

    2015-01-01

    Huntington’s disease is an autosomal dominant disease associated with a mutation in the gene encoding huntingtin (Htt) leading to expanded polyglutamine repeats of mutant Htt (mHtt) that elicit oxidative stress, neurotoxicity, and motor and behavioural changes1. Huntington’s disease is characterized by highly selective and profound damage to the corpus striatum, which regulates motor function. Striatal selectivity of Huntington’s disease may reflect the striatally selective small G protein Rhes binding to mHtt and enhancing its neurotoxicity2. Specific molecular mechanisms by which mHtt elicits neurodegeneration have been hard to determine. Here we show a major depletion of cystathionine γ-lyase (CSE), the biosynthetic enzyme for cysteine, in Huntington’s disease tissues, which may mediate Huntington’s disease pathophysiology. The defect occurs at the transcriptional level and seems to reflect influences of mHtt on specificity protein 1, a transcriptional activator for CSE. Consistent with the notion of loss of CSE as a pathogenic mechanism, supplementation with cysteine reverses abnormalities in cultures of Huntington’s disease tissues and in intact mouse models of Huntington’s disease, suggesting therapeutic potential. PMID:24670645

  14. Finding the Subcellular Location of Barley, Wheat, Rice and Maize Proteins: The Compendium of Crop Proteins with Annotated Locations (cropPAL).

    Science.gov (United States)

    Hooper, Cornelia M; Castleden, Ian R; Aryamanesh, Nader; Jacoby, Richard P; Millar, A Harvey

    2016-01-01

    Barley, wheat, rice and maize provide the bulk of human nutrition and have extensive industrial use as agricultural products. The genomes of these crops each contains >40,000 genes encoding proteins; however, the major genome databases for these species lack annotation information of protein subcellular location for >80% of these gene products. We address this gap, by constructing the compendium of crop protein subcellular locations called crop Proteins with Annotated Locations (cropPAL). Subcellular location is most commonly determined by fluorescent protein tagging of live cells or mass spectrometry detection in subcellular purifications, but can also be predicted from amino acid sequence or protein expression patterns. The cropPAL database collates 556 published studies, from >300 research institutes in >30 countries that have been previously published, as well as compiling eight pre-computed subcellular predictions for all Hordeum vulgare, Triticum aestivum, Oryza sativa and Zea mays protein sequences. The data collection including metadata for proteins and published studies can be accessed through a search portal http://crop-PAL.org. The subcellular localization information housed in cropPAL helps to depict plant cells as compartmentalized protein networks that can be investigated for improving crop yield and quality, and developing new biotechnological solutions to agricultural challenges. © The Author 2015. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oup.com.

  15. Positive effects of temperature and growth conditions on enzymatic and antioxidant status in lettuce plants.

    Science.gov (United States)

    Boo, Hee-Ock; Heo, Buk-Gu; Gorinstein, Shela; Chon, Sang-Uk

    2011-10-01

    The contents of two bioactive compounds (polyphenols and flavonoids) and their antioxidant and enzyme activities were determined in the leaves of six lettuce (Latuca sativa L.) cultivars subjected to 4 different day/night temperatures for 6 weeks. The total polyphenol and anthocyanin contents and the corresponding antioxidant activities were the highest at 13/10°C and 20/13°C, followed by 25/20°C and 30/25°C. The enzymatic activities of polyphenol oxidase (PPO) and phenylalanine ammonia-lyase (PAL) were also the highest at low day/night temperatures, but the peroxidase (POD) activity was decreased at low day/night temperatures and increased at high day/night temperatures. The most significant positive correlation existed between anthocyanin content and PPO activity, total polyphenols and their antioxidant activities. The results showed that at relatively low temperatures, lettuce plants have a high antioxidant and enzymatic status. These results provide additional information for the lettuce growers. Copyright © 2011 Elsevier Ireland Ltd. All rights reserved.

  16. Experimental and theoretical investigations of mechanisms responsible for plasma jets formation at PALS

    Czech Academy of Sciences Publication Activity Database

    Kasperczuk, A.; Pisarczyk, T.; Demchenko, N. N.; Gus'kov, S. Yu.; Kálal, M.; Ullschmied, Jiří; Krouský, Eduard; Mašek, Karel; Pfeifer, Miroslav; Rohlena, Karel; Skála, Jiří; Pisarczyk, P.

    2009-01-01

    Roč. 27, č. 3 (2009), s. 415-427 ISSN 0263-0346 R&D Projects: GA MŠk(CZ) LC528 Institutional research plan: CEZ:AV0Z20430508; CEZ:AV0Z10100523 Keywords : Laser-plasma interaction * laser produced-plasma jet * radiative cooling * target irradiation geometry * PALS laser * laser interferometry Subject RIV: BL - Plasma and Gas Discharge Physics Impact factor: 4.420, year: 2008

  17. Mode of action of pectin lyase A of Aspergillus niger on differently C6-substituted oligogalacturonides

    NARCIS (Netherlands)

    Alebeek, van G.J.W.M.; Christensen, T.M.I.E.; Schols, H.A.; Mikkelsen, J.D.; Voragen, A.G.J.

    2002-01-01

    A thorough investigation of the mode of action of Aspergillus niger (4M-147) pectin lyase A (PLA) on differently C6-substituted oligogalacturonides is described. PLA appeared to be very specific for fully methyl-esterified oligogalacturonides: removal of the methyl-ester or changing the type of

  18. Kinetic Resolution and Stereoselective Synthesis of 3-Substituted Aspartic Acids by Using Engineered Methylaspartate Ammonia Lyases

    NARCIS (Netherlands)

    Raj, Hans; Szymanski, Wiktor; Villiers, Jandré de; Puthan Veetil, Vinod; Quax, Wim J.; Shimamoto, Keiko; Janssen, Dick B.; Feringa, Ben L.; Poelarends, Gerrit J.

    2013-01-01

    Kinetic resolution and asymmetric synthesis of various valuable 3-substituted aspartic acids, which were obtained in fair to good yields with diastereomeric ratio values of up to >98:2 and enantiomeric excess values of up to >99 %, by using engineered methylaspartate ammonia lyases are described.

  19. Feasibility of Radiation Processing for Post-Harvest Storage of Potatoes Under Tropical Conditions

    Energy Technology Data Exchange (ETDEWEB)

    Thomas, P.; Srirangarajan, A. N.; Padwal-Desai, S. R.; Ghanekar, A. S.; Shirsat, S. G.; Pendharkar, M. B.; Nair, P. M.; Nadkarni, G. B. [Biochemistry and Food Technology Division, Bhabha Atomic Research Centre, Bombay (India)

    1978-04-15

    Storage changes after gamma irradiation (10 krad) of five commercial varieties of potatoes, Kufri Chandramukhi, K. Kuber, K. Alankar, K. Sheetman and K. Sinduri, grown in India were ascertained to arrive at optimal storage conditions. The radiation treatment could completely inhibit sprouting in all the varieties, irrespective of storage temperature, in addition to eliminating the tuber moth Pthorimaea operculella (Zeller). The temperature-dependent total losses could be attributed to microbial rotting and dehydration. The organisms causing soft rot at temperatures above 15 Degree-Sign C belonged to Erwinia spp., while the rot caused below 10 Degree-Sign C was essentially due to Micrococcus spp. Irradiation followed by storage at cool temperatures (10-15 Degree-Sign C) offers an alternative method, comparable with conventional refrigerated (2-4 Degree-Sign C) storage. The irradiated potatoes were not amenable to the attack of toxigenic fungi Aspergillus flavus and A. parasiticus during storage. This could be attributed to phenolic compounds present in the tubers. Phenylalanine ammonia lyase (PAL), involved in the metabolism of phenolic compounds, showed enhancement in the irradiated potatoes. The extent of radiation induced activation of PAL is suggested as a biochemical parameter for identifying irradiated tubers. (author)

  20. Feasibility of radiation processing for post-harvest storage of potatoes under tropical conditions

    International Nuclear Information System (INIS)

    Thomas, P.; Srirangarajan, A.N.; Padwal-Desai, S.R.; Ghanekar, A.S.; Shirsat, S.G.; Pendharkar, M.B.; Nair, P.M.; Nadkarni, G.B.

    1978-01-01

    Storage changes after gamma irradiation (10krad) of five commercial varieties of potatoes, Kufri Chandramukhi, K. Kuber, K. Alankar, K. Sheetman and K. Sinduri, grown in India were ascertained to arrive at optimal storage conditions. The radiation treatment could completely inhibit sprouting in all the varieties, irrespective of storage temperature, in addition to eliminating the tuber moth Pthorimaea operculella (Zeller). The temperature-dependent total losses could be attributed to microbial rotting and dehydration. The organisms causing soft rot at temperatures above 15 0 C belonged to Erwinia spp., while the rot caused below 10 0 C was essentially due to Micrococcus spp. Irradiation followed by storage at cool temperatures (10-15 0 C) offers an alternative method, comparable with conventional refrigerated (2-4 0 C) storage. The irradiated potatoes were not amenable to the attack of toxigenic fungi Aspergillus flavus and A. parasiticus during storage. This could be attributed to phenolic compounds present in the tubers. Phenylalanine ammonia lyase (PAL), involved in the metabolism of phenolic compounds, showed enhancement in the irradiated potatoes. The extent of radiation-induced activation of PAL is suggested as a biochemical parameter for identifying irradiated tubers. (author)

  1. Characterization of control mesoporous glasses (CPGs) using positron annihilation lifetime spectroscopy (PALS)

    Energy Technology Data Exchange (ETDEWEB)

    Sayed Ahmed, Essmat Mahmoud Hassan

    2008-01-30

    This thesis has two main goals. Firstly and for the first time, the positron annihilation lifetime technique (PALS) is used to characterize the control porous glasses (CPGs) media. All the positron annihilation spectroscopy (PAS) investigations have been interested in the commercial Vycor glass media of pores size {<=} 4 nm. Therefore, the PALS is used to establish basic correlations between the important physical properties of the CPG (pore size, surface area, and porosity) and the orthopositronium (o-Ps) lifetime. The second goal is to use these correlations to verify the validity of some suggested models and theories to discover possible deviations from the expected behaviour and to discuss the physical point of view for these deviations. Chapter 1 presents an overview of positron, positronium and their interaction with solid materials. Chapter 2 of this thesis is concerned with characterization of porous glass materials. This chapter discusses in detail the preparation methods of porous glasses specially control porous glass and Vycor glass. The gas adsorption phenomenon in porous glass and the related definitions are discussed. The most important terms of the porous glass such as pore size, shape, volume, pore size distribution, surface area, and porosity are discussed. Chapter 3 presents in detail the well known models and the derived equations which correlate between the lifetime of the o-Ps long-lived component and the pore size, pore shape and the temperature dependence of the porous media. Chapter 4 presents an overview of the main experimental techniques used in this thesis, namely positron annihilation lifetime spectroscopy. Chapter 5 explains the sample preparation for the measurements. The chapter represents also the experimental data for the correlation between the o-Ps lifetime and pore sizes for a system of CPG media (from 1 nm to 64 nm). I also studied the temperature dependence of the o-Ps long-lived component in different pore sizes. In this

  2. Characterization of control mesoporous glasses (CPGs) using positron annihilation lifetime spectroscopy (PALS)

    International Nuclear Information System (INIS)

    Sayed Ahmed, Essmat Mahmoud Hassan

    2008-01-01

    This thesis has two main goals. Firstly and for the first time, the positron annihilation lifetime technique (PALS) is used to characterize the control porous glasses (CPGs) media. All the positron annihilation spectroscopy (PAS) investigations have been interested in the commercial Vycor glass media of pores size ≤ 4 nm. Therefore, the PALS is used to establish basic correlations between the important physical properties of the CPG (pore size, surface area, and porosity) and the orthopositronium (o-Ps) lifetime. The second goal is to use these correlations to verify the validity of some suggested models and theories to discover possible deviations from the expected behaviour and to discuss the physical point of view for these deviations. Chapter 1 presents an overview of positron, positronium and their interaction with solid materials. Chapter 2 of this thesis is concerned with characterization of porous glass materials. This chapter discusses in detail the preparation methods of porous glasses specially control porous glass and Vycor glass. The gas adsorption phenomenon in porous glass and the related definitions are discussed. The most important terms of the porous glass such as pore size, shape, volume, pore size distribution, surface area, and porosity are discussed. Chapter 3 presents in detail the well known models and the derived equations which correlate between the lifetime of the o-Ps long-lived component and the pore size, pore shape and the temperature dependence of the porous media. Chapter 4 presents an overview of the main experimental techniques used in this thesis, namely positron annihilation lifetime spectroscopy. Chapter 5 explains the sample preparation for the measurements. The chapter represents also the experimental data for the correlation between the o-Ps lifetime and pore sizes for a system of CPG media (from 1 nm to 64 nm). I also studied the temperature dependence of the o-Ps long-lived component in different pore sizes. In this

  3. Transcriptional regulation of Saccharomyces cerevisiaeCYS3 encoding cystathionine γ-lyase

    Science.gov (United States)

    Hiraishi, Hiroyuki; Miyake, Tsuyoshi

    2008-01-01

    In studying the regulation of GSH11, the structural gene of the high-affinity glutathione transporter (GSH-P1) in Saccharomyces cerevisiae, a cis-acting cysteine responsive element, CCGCCACAC (CCG motif), was detected. Like GSH-P1, the cystathionine γ-lyase encoded by CYS3 is induced by sulfur starvation and repressed by addition of cysteine to the growth medium. We detected a CCG motif (−311 to −303) and a CGC motif (CGCCACAC; −193 to −186), which is one base shorter than the CCG motif, in the 5′-upstream region of CYS3. One copy of the centromere determining element 1, CDE1 (TCACGTGA; −217 to −210), being responsible for regulation of the sulfate assimilation pathway genes, was also detected. We tested the roles of these three elements in the regulation of CYS3. Using a lacZ-reporter assay system, we found that the CCG/CGC motif is required for activation of CYS3, as well as for its repression by cysteine. In contrast, the CDE1 motif was responsible for only activation of CYS3. We also found that two transcription factors, Met4 and VDE, are responsible for activation of CYS3 through the CCG/CGC and CDE1 motifs. These observations suggest a dual regulation of CYS3 by factors that interact with the CDE1 motif and the CCG/CGC motifs. PMID:18317767

  4. Enzyme Activities in Oleaginous Yeasts Accumulating and Utilizing Exogenous or Endogenous Lipids

    NARCIS (Netherlands)

    Holdsworth, Jane E.; Veenhuis, Marten; Ratledge, Colin

    1988-01-01

    The activities of ATP:citrate lyase (ACL; EC 4.1.3.8), carnitine acetyltransferase (CAT; EC 2.3.1.7), NADP+-dependent isocitrate dehydrogenase (ICDH; EC 1.1.1.42), isocitrate lyase (ICL; EC 4.1.3.1) and malic enzyme (malate dehydrogenase; EC 1.1.1.40) were measured in four oleaginous yeasts, Candida

  5. Comparison of expression, purification and characterization of a new pectate lyase from Phytophthora capsici using two different methods

    Directory of Open Access Journals (Sweden)

    Zhang Xiuguo

    2011-04-01

    Full Text Available Abstract Background Pectate lyases (PELs play an important role in the infection process of plant pathogens and also have a commercial significance in industrial applications. Most of the PELs were expressed as soluble recombinant proteins, while a few recombinant proteins were insoluble. The production of a large-scale soluble recombinant PEL would allow not only a more detailed structural and functional characterization of this enzyme but also may have important applications in the food industry. Results We cloned a new pectate lyase gene (Pcpel2 from Phytophthora capsici. Pcpel2 was constructed by pET system and pMAL system, and both constructs were used to express the PCPEL2 in Escherichia coli BL21 (DE3 pLysS. The expressed products were purified using affinity chromatography and gel filtration chromatography. The purity, specific activity and pathogenicity of the purified PCPEL2 expressed by the pMAL system were higher than the purified PCPEL2 expressed by the pET system. In addition, some other characteristics of the purified PCPEL2 differed from the two systems, such as crystallographic features. Purified PCPEL2 expressed by the pMAL system was crystallized by the hanging-drop vapour-diffusion method at 289 K, and initial crystals were grown. Conclusion The two different methods and comparison presented here would be highly valuable in obtaining an ideal enzyme for the downstream experiments, and supply an useful alternative to purify some insoluble recombinant proteins.

  6. Pseudomonas aeruginosa 4-amino-4-deoxychorismate lyase: spatial conservation of an active site tyrosine and classification of two types of enzyme.

    Directory of Open Access Journals (Sweden)

    Patrick E F O'Rourke

    Full Text Available 4-Amino-4-deoxychorismate lyase (PabC catalyzes the formation of 4-aminobenzoate, and release of pyruvate, during folate biosynthesis. This is an essential activity for the growth of gram-negative bacteria, including important pathogens such as Pseudomonas aeruginosa. A high-resolution (1.75 Å crystal structure of PabC from P. aeruginosa has been determined, and sequence-structure comparisons with orthologous structures are reported. Residues around the pyridoxal 5'-phosphate cofactor are highly conserved adding support to aspects of a mechanism generic for enzymes carrying that cofactor. However, we suggest that PabC can be classified into two groups depending upon whether an active site and structurally conserved tyrosine is provided from the polypeptide that mainly forms an active site or from the partner subunit in the dimeric assembly. We considered that the conserved tyrosine might indicate a direct role in catalysis: that of providing a proton to reduce the olefin moiety of substrate as pyruvate is released. A threonine had previously been suggested to fulfill such a role prior to our observation of the structurally conserved tyrosine. We have been unable to elucidate an experimentally determined structure of PabC in complex with ligands to inform on mechanism and substrate specificity. Therefore we constructed a computational model of the catalytic intermediate docked into the enzyme active site. The model suggests that the conserved tyrosine helps to create a hydrophobic wall on one side of the active site that provides important interactions to bind the catalytic intermediate. However, this residue does not appear to participate in interactions with the C atom that undergoes an sp(2 to sp(3 conversion as pyruvate is produced. The model and our comparisons rather support the hypothesis that an active site threonine hydroxyl contributes a proton used in the reduction of the substrate methylene to pyruvate methyl in the final stage of

  7. PecS and PecT coregulate the synthesis of HrpN and pectate lyases, two virulence determinants in Erwinia chrysanthemi 3937.

    Science.gov (United States)

    Nasser, William; Reverchon, Sylvie; Vedel, Regine; Boccara, Martine

    2005-11-01

    Erwinia chrysanthemi strain 3937 is a necrotrophic bacterial plant pathogen. Pectinolytic enzymes and, in particular, pectate lyases play a key role in soft rot symptoms; however, the efficient colonization of plants by E. chrysanthemi requires additional factors. These factors include HrpN (harpin), a heat-stable, glycine-rich hydrophilic protein, which is secreted by the type III secretion system. We investigated the expression of hrpN in E. chrysanthemi 3937 in various environmental conditions and different regulatory backgrounds. Using lacZ fusions, hrpN expression was markedly influenced by the carbon source, osmolarity, growth phase, and growth substrate. hrpN was repressed when pectinolysis started and negatively regulated by the repressors of pectate lyase synthesis, PecS and PecT. Primer extension data and in vitro DNA-protein interaction experiments support a model whereby PecS represses hrpN expression by binding to the hrpN regulatory region and inhibiting transcript elongation. The results suggest coordinated regulation of HrpN and pectate lyases by PecS and PecT. A putative model of the synthesis of these two virulence factors in E. chrysanthemi during pathogenesis is presented.

  8. Free volume changes in mechanically milled PS and PC studied by positron annihilation lifetime spectroscopy (PALS)

    NARCIS (Netherlands)

    Günther-Schade, K.; Castricum, H.L.; Ziegler, H.J.; Bakker, H.; Faupel, F.

    2004-01-01

    The effect of mechanical milling on free volume was studied by means of positron annihilation lifetime spectroscopy (PALS) in polystyrene (PS) as a typical brittle polymer and in polycarbonate (PC) as a tough representative. Long-time milling increases the free volume, while a decrease is observed

  9. Azimuthal Signature of Coincidental Brightness Temperature and Normalized Radar Cross-Section Obtained Using Airborne PALS Instrument

    Science.gov (United States)

    Colliander, Andreas; Kim, Seungbum; Yueh, Simon; Cosh, Mike; Jackson, Tom; Njoku, Eni

    2010-01-01

    Coincidental airborne brightness temperature (TB) and normalized radar-cross section (NRCS) measurements were carried out with the PALS (Passive and Active L- and S-band) instrument in the SMAPVEX08 (SMAP Validation Experiment 2008) field campaign. This paper describes results obtained from a set of flights which measured a field in 45(sup o) steps over the azimuth angle. The field contained mature soy beans with distinct row structure. The measurement shows that both TB and NRCS experience modulation effects over the azimuth as expected based on the theory. The result is useful in development and validation of land surface parameter forward models and retrieval algorithms, such as the soil moisture algorithm for NASA's SMAP (Soil Moisture Active and Passive) mission. Although the footprint of the SMAP will not be sensitive to the small resolution scale effects as the one presented in this paper, it is nevertheless important to understand the effects at smaller scale.

  10. Influence of anthropometric factors and physical activity on blood pressure in adolescents from Taguatinga, Federal District, Brazil

    Directory of Open Access Journals (Sweden)

    Sebastião Lobo Silva

    2013-04-01

    Full Text Available This study aimed to analyze the influence of body mass index (BMI and physical activity level (PAL on blood pressure (BP in adolescents. The sample was composed of 799 students of both genders, aged 12.09 (± 0.80. The weight, height and BP were assessed according to internationally established criteria, and BMI was calculated and classified according to the cutoff points proposed by Cole, Flegal and Dietz (2000. To evaluate the PAL, the modified lifestyle questionnaire was used (Silva, Silva, & Martins, 2006. We found that 9.1% of the subjects evaluated have high blood pressure, 11.8% were overweight, 5.4% were obese and 47.2% of the people presented low PAL. The fact of being inactive (OR = 1.99, insufficiently active (OR = 1.87 and obese (OR = 5.1 was associated with an increased risk of developing hypertension. Being inactive or insufficiently active was strongly associated with the development of the obesity (OR = 7.97 and 4.35 respectively. In the studied sample, the overweight, the obesity and low PAL are associated with the development of arterial hypertension. In addition, a low PAL is associated with an increasing number of overweight adolescents.

  11. Mitochondrial pAL2-1 plasmid homologs are senescence factors in Podospora anserina independent of intrinsic senescence

    NARCIS (Netherlands)

    Diepeningen, van A.D.; Debets, A.J.M.; Slakhorst-Wandel, S.M.; Hoekstra, R.F.

    2008-01-01

    Since the first description of a linear mitochondrial plasmid in Podospora anserina, pAL2-1, and homologous plasmids have gone from being considered beneficial longevity plasmids, via neutral genetic elements, toward mutator plasmids causing senescence. The plasmid has an invertron structure, with

  12. Mitochondrial pAL2-1 plasmid homologs are senescence factors in Podospora anserina independent of intrinsic senescence

    NARCIS (Netherlands)

    van Diepeningen, Anne D; Debets, Alfons J M; Slakhorst, S Marijke; Hoekstra, Rolf F

    Since the first description of a linear mitochondrial plasmid in Podospora anserina, pAL2-1, and homologous plasmids have gone from being considered beneficial longevity plasmids, via neutral genetic elements, toward mutator plasmids causing senescence. The plasmid has an invertron structure, with

  13. Water-Vapor Sorption Processes in Nanoporous MgO-Al2O3 Ceramics: the PAL Spectroscopy Study.

    Science.gov (United States)

    Klym, Halyna; Ingram, Adam; Shpotyuk, Oleh; Hadzaman, Ivan; Solntsev, Viacheslav

    2016-12-01

    The water-vapor sorption processes in nanoporous MgO-Al2O3 ceramics are studied with positron annihilation lifetime (PAL) spectroscopy employing positron trapping and positronium (Ps)-decaying modes. It is demonstrated that the longest-lived components in the four-term reconstructed PAL spectra with characteristic lifetimes near 2 and 60-70 ns can be, respectively, attributed to ortho-positronium (o-Ps) traps in nanopores with 0.3- and 1.5-1.8-nm radii. The first o-Ps decaying process includes "pick-off" annihilation in the "bubbles" of liquid water, while the second is based on o-Ps interaction with physisorbed water molecules at the walls of the pores. In addition, the water vapor modifies structural defects located at the grain boundaries in a vicinity of pores, this process being accompanied by void fragmentation during water adsorption and agglomeration during water desorption after drying.

  14. Water-Vapor Sorption Processes in Nanoporous MgO-Al2O3 Ceramics: the PAL Spectroscopy Study

    Science.gov (United States)

    Klym, Halyna; Ingram, Adam; Shpotyuk, Oleh; Hadzaman, Ivan; Solntsev, Viacheslav

    2016-03-01

    The water-vapor sorption processes in nanoporous MgO-Al2O3 ceramics are studied with positron annihilation lifetime (PAL) spectroscopy employing positron trapping and positronium (Ps)-decaying modes. It is demonstrated that the longest-lived components in the four-term reconstructed PAL spectra with characteristic lifetimes near 2 and 60-70 ns can be, respectively, attributed to ortho-positronium (o-Ps) traps in nanopores with 0.3- and 1.5-1.8-nm radii. The first o-Ps decaying process includes "pick-off" annihilation in the "bubbles" of liquid water, while the second is based on o-Ps interaction with physisorbed water molecules at the walls of the pores. In addition, the water vapor modifies structural defects located at the grain boundaries in a vicinity of pores, this process being accompanied by void fragmentation during water adsorption and agglomeration during water desorption after drying.

  15. Insights into the structural characteristics and substrate binding analysis of chondroitin AC lyase (PsPL8A) from Pedobacter saltans.

    Science.gov (United States)

    Rani, Aruna; Dhillon, Arun; Sharma, Kedar; Goyal, Arun

    2018-04-01

    The structure of chondroitin AC lyase (PsPL8A) of family 8 polysaccharide lyase was characterized. Modeled PsPL8A structure showed, it contains N-terminal (α/α) 6 incomplete toroidal fold and a layered β sandwich structure at C-terminal. Ramchandran plot displayed 98.5% residues in favoured and 1.2% in generously allowed region. Secondary structure of PsPL8A by CD revealed 27.31% α helices 22.7% β sheets and 49.9% random coils. Protein melting study showed, PsPL8A completely unfolds at 60°C. SAXS analysis showed, PsPL8A is fully folded in solution form. The ab initio derived dummy model of PsPL8A superposed well with its modeled structure excluding some α-helices and loop region. Structural superposition and docking analysis showed, N153, W105, H203, Y208, Y212, R266 and E349 were involved in catalysis. Mutants N153A, H203A, Y212F, R266A and E349A created by SDM revealed no residual activity. Isothermal titration calorimetry analysis of Y212F and H203A with C4S polysaccharide, showed moderate binding by Y212F (Ka=9.56±3.81×10 5 ) and no binding with H203A, showing active contribution of Y212 in substrate binding. Residues Y212 and H203 or R266 might act as general base and general acid respectively. Residues N153 and E349 are likely contributing in charge neutralization and stabilizing enolate anion intermediate during β-elimination. Copyright © 2017 Elsevier B.V. All rights reserved.

  16. Thyme oil vapour and modified atmosphere packaging reduce anthracnose incidence and maintain fruit quality in avocado.

    Science.gov (United States)

    Sellamuthu, Periyar Selvam; Mafune, Mpho; Sivakumar, Dharini; Soundy, Puffy

    2013-09-01

    Postharvest application of prochloraz fungicide is commercially practiced to control anthracnose, a postharvest disease in avocado. Increasing consumer concern regarding food safety and demand for organically produced fruits make it necessary to search for natural environmentally friendly alternative products and processes for the fruit industry. A combination of modified atmosphere packaging (MAP; ∼8% CO₂, 2% O₂) plus thyme oil (TO) was evaluated on the incidence and severity of anthracnose, physiological disorders (grey pulp, vascular browning), fruit quality parameters (L*, h°, firmness, weight loss) and sensory parameters (taste, texture, flavour and overall acceptance), phenylalanine ammonia-lyase (PAL) enzyme activity, total phenolic compounds, flavonoid contents and antioxidant activity in avocados ('Fuerte' and 'Hass' cultivars) held at 10 °C cold storage for 18 days and thereafter, ripened at 25 °C for 5-10 days. Stand-alone MAP, commercial treatment (prochloraz 0.05%) and untreated (control) fruit were included for comparison. MAP + TO treatment significantly (P anthracnose, grey pulp, vascular browning, weight loss and loss of fruit firmness, and showed acceptable taste, flavour, texture and higher overall acceptance, increased PAL activity, total phenolic compounds, flavonoid contents and antioxidant activity, after ripening at 25 °C followed by cold storage at 10 °C. This investigation recommends MAP + TO combination treatment as a suitable alternative to the currently adopted prochloraz application. © 2013 Society of Chemical Industry.

  17. Sphingosine-1-phosphate (S1P) displays sustained S1P1 receptor agonism and signaling through S1P lyase-dependent receptor recycling.

    Science.gov (United States)

    Gatfield, John; Monnier, Lucile; Studer, Rolf; Bolli, Martin H; Steiner, Beat; Nayler, Oliver

    2014-07-01

    The sphingosine-1-phosphate (S1P) type 1 receptor (S1P1R) is a novel therapeutic target in lymphocyte-mediated autoimmune diseases. S1P1 receptor desensitization caused by synthetic S1P1 receptor agonists prevents T-lymphocyte egress from secondary lymphoid organs into the circulation. The selective S1P1 receptor agonist ponesimod, which is in development for the treatment of autoimmune diseases, efficiently reduces peripheral lymphocyte counts and displays efficacy in animal models of autoimmune disease. Using ponesimod and the natural ligand S1P, we investigated the molecular mechanisms leading to different signaling, desensitization and trafficking behavior of S1P1 receptors. In recombinant S1P1 receptor-expressing cells, ponesimod and S1P triggered Gαi protein-mediated signaling and β-arrestin recruitment with comparable potency and efficiency, but only ponesimod efficiently induced intracellular receptor accumulation. In human umbilical vein endothelial cells (HUVEC), ponesimod and S1P triggered translocation of the endogenous S1P1 receptor to the Golgi compartment. However, only ponesimod treatment caused efficient surface receptor depletion, receptor accumulation in the Golgi and degradation. Impedance measurements in HUVEC showed that ponesimod induced only short-lived Gαi protein-mediated signaling followed by resistance to further stimulation, whereas S1P induced sustained Gαi protein-mediated signaling without desensitization. Inhibition of S1P lyase activity in HUVEC rendered S1P an efficient S1P1 receptor internalizing compound and abrogated S1P-mediated sustained signaling. This suggests that S1P lyase - by facilitating S1P1 receptor recycling - is essential for S1P-mediated sustained signaling, and that synthetic agonists are functional antagonists because they are not S1P lyase substrates. Copyright © 2014 Elsevier Inc. All rights reserved.

  18. Systematic review of acute physically active learning and classroom movement breaks on children's physical activity, cognition, academic performance and classroom behaviour: understanding critical design features.

    Science.gov (United States)

    Daly-Smith, Andy J; Zwolinsky, Stephen; McKenna, Jim; Tomporowski, Phillip D; Defeyter, Margaret Anne; Manley, Andrew

    2018-01-01

    To examine the impact of acute classroom movement break (CMB) and physically active learning (PAL) interventions on physical activity (PA), cognition, academic performance and classroom behaviour. Systematic review. PubMed, EBSCO, Academic Search Complete, Education Resources Information Center, PsycINFO, SPORTDiscus, SCOPUS and Web of Science. Studies investigating school-based acute bouts of CMB or PAL on (PA), cognition, academic performance and classroom behaviour. The Downs and Black checklist assessed risk of bias. Ten PAL and eight CMB studies were identified from 2929 potentially relevant articles. Risk of bias scores ranged from 33% to 64.3%. Variation in study designs drove specific, but differing, outcomes. Three studies assessed PA using objective measures. Interventions replaced sedentary time with either light PA or moderate-to-vigorous PA dependent on design characteristics (mode, duration and intensity). Only one study factored individual PA outcomes into analyses. Classroom behaviour improved after longer moderate-to-vigorous (>10 min), or shorter more intense (5 min), CMB/PAL bouts (9 out of 11 interventions). There was no support for enhanced cognition or academic performance due to limited repeated studies. Low-to-medium quality designs predominate in investigations of the acute impacts of CMB and PAL on PA, cognition, academic performance and classroom behaviour. Variable quality in experimental designs, outcome measures and intervention characteristics impact outcomes making conclusions problematic. CMB and PAL increased PA and enhanced time on task. To improve confidence in study outcomes, future investigations should combine examples of good practice observed in current studies. CRD42017070981.

  19. Enhanced Arabidopsis disease resistance against Botrytis cinerea induced by sulfur dioxide.

    Science.gov (United States)

    Xue, Meizhao; Yi, Huilan

    2018-01-01

    Sulfur dioxide (SO 2 ) is a common air pollutant that has complex impacts on plants. The effect of prior exposure to 30mgm -3 SO 2 on defence against Botrytis cinerea (B. cinerea) in Arabidopsis thaliana and the possible mechanisms of action were investigated. The results indicated that pre-exposure to 30mgm -3 SO 2 resulted in significantly enhanced resistance to B. cinerea infection. SO 2 pre-treatment significantly enhanced the activities of defence-related enzymes including phenylalanine ammonia-lyase (PAL), polyphenol oxidase (PPO), β-1,3-glucanase (BGL) and chitinase (CHI). Transcripts of the defence-related genes PAL, PPO, PR2, and PR3, encoding PAL, PPO, BGL and CHI, respectively, were markedly elevated in Arabidopsis plants pre-exposed to SO 2 and subsequently inoculated with B. cinerea (SO 2 + treatment group) compared with those that were only treated with SO 2 (SO 2 ) or inoculated with B. cinerea (CK+). Moreover, SO 2 pre-exposure also led to significant increases in the expression levels of MIR393, MIR160 and MIR167 in Arabidopsis. Meanwhile, the expression of known targets involved in the auxin signalling pathway, was negatively correlated with their corresponding miRNAs. Additionally, the transcript levels of the primary auxin-response genes GH3-like, BDL/IAA12, and AXR3/IAA17 were markedly repressed. Our findings indicate that 30mgm -3 SO 2 pre-exposure enhances disease resistance against B. cinerea in Arabidopsis by priming defence responses through enhancement of defence-related gene expression and enzyme activity, and miRNA-mediated suppression of the auxin signalling pathway. Copyright © 2017 Elsevier Inc. All rights reserved.

  20. The Richmond Agitation-Sedation Scale modified for palliative care inpatients (RASS-PAL): a pilot study exploring validity and feasibility in clinical practice.

    Science.gov (United States)

    Bush, Shirley H; Grassau, Pamela A; Yarmo, Michelle N; Zhang, Tinghua; Zinkie, Samantha J; Pereira, José L

    2014-03-31

    The Richmond Agitation-Sedation Scale (RASS), which assesses level of sedation and agitation, is a simple observational instrument which was developed and validated for the intensive care setting. Although used and recommended in palliative care settings, further validation is required in this patient population. The aim of this study was to explore the validity and feasibility of a version of the RASS modified for palliative care populations (RASS-PAL). A prospective study, using a mixed methods approach, was conducted. Thirteen health care professionals (physicians and nurses) working in an acute palliative care unit assessed ten consecutive patients with an agitated delirium or receiving palliative sedation. Patients were assessed at five designated time points using the RASS-PAL. Health care professionals completed a short survey and data from semi-structured interviews was analyzed using thematic analysis. The inter-rater intraclass correlation coefficient range of the RASS-PAL was 0.84 to 0.98 for the five time points. Professionals agreed that the tool was useful for assessing sedation and was easy to use. Its role in monitoring delirium however was deemed problematic. Professionals felt that it may assist interprofessional communication. The need for formal education on why and how to use the instrument was highlighted. This study provides preliminary validity evidence for the use of the RASS-PAL by physicians and nurses working in a palliative care unit, specifically for assessing sedation and agitation levels in the management of palliative sedation. Further validity evidence should be sought, particularly in the context of assessing delirium.

  1. The Richmond Agitation-Sedation Scale modified for palliative care inpatients (RASS-PAL): a pilot study exploring validity and feasibility in clinical practice

    Science.gov (United States)

    2014-01-01

    Background The Richmond Agitation-Sedation Scale (RASS), which assesses level of sedation and agitation, is a simple observational instrument which was developed and validated for the intensive care setting. Although used and recommended in palliative care settings, further validation is required in this patient population. The aim of this study was to explore the validity and feasibility of a version of the RASS modified for palliative care populations (RASS-PAL). Methods A prospective study, using a mixed methods approach, was conducted. Thirteen health care professionals (physicians and nurses) working in an acute palliative care unit assessed ten consecutive patients with an agitated delirium or receiving palliative sedation. Patients were assessed at five designated time points using the RASS-PAL. Health care professionals completed a short survey and data from semi-structured interviews was analyzed using thematic analysis. Results The inter-rater intraclass correlation coefficient range of the RASS-PAL was 0.84 to 0.98 for the five time points. Professionals agreed that the tool was useful for assessing sedation and was easy to use. Its role in monitoring delirium however was deemed problematic. Professionals felt that it may assist interprofessional communication. The need for formal education on why and how to use the instrument was highlighted. Conclusion This study provides preliminary validity evidence for the use of the RASS-PAL by physicians and nurses working in a palliative care unit, specifically for assessing sedation and agitation levels in the management of palliative sedation. Further validity evidence should be sought, particularly in the context of assessing delirium. PMID:24684942

  2. Interferometric investigation of an early stage of plasma expansion with the high-power laser system PALS

    Czech Academy of Sciences Publication Activity Database

    Kasperczuk, A.; Pisarczyk, P.; Pisarczyk, T.; Králiková, Božena; Mašek, Karel; Pfeifer, Miroslav; Rohlena, Karel; Skála, Jiří; Ullschmied, Jiří; Kálal, M.

    2002-01-01

    Roč. 52, č. 3 (2002), s. 395-404 ISSN 0011-4626 R&D Projects: GA MŠk LN00A100 Institutional research plan: CEZ:AV0Z2043910 Keywords : laser plasma, PALS laser system, laser interferometry Subject RIV: BH - Optics, Masers, Lasers Impact factor: 0.311, year: 2002

  3. Increased protein expression of LHCG receptor and 17a-hydroxylase/17,20-lyase in human polycystic ovaries

    NARCIS (Netherlands)

    Comim, F.V.; Teerds, K.J.; Hardy, K.; Franks, S.

    2013-01-01

    STUDY QUESTION Does the expression of LHCG receptor (LHCGR) protein and key enzymes in the androgen biosynthetic pathway differ in normal human versus polycystic ovarian tissue? SUMMARY ANSWER LHCGR and 17a-hydroxylase/17-20-lyase (CYP17A1) protein levels are increased in polycystic ovaries (PCOs).

  4. Metabolically engineered cells for the production of resveratrol or an oligomeric or glycosidically-bound derivative thereof

    DEFF Research Database (Denmark)

    2006-01-01

    A from said 4-coumaric acid, and resveratrol synthase (VST) produces said resveratrol from said 4- coumaroyl CoA, or in which L-phenylalanine- or tyrosine- ammonia lyase (PAL/TAL) produces 4-coumaric acid, 4- coumarate-CoA ligase (4CL) produces 4-coumaroyl CoA from said 4-coumaric acid, and resveratrol...... synthase (VST) produces said resveratrol from said 4-coumaroyl CoA. The micro-organism may be a yeast, fungus or bacterium including Saccharomyces cerevisiae, E. coli, Lactococcus lactis, Aspergillus niger, or Aspergillus oryzae....

  5. Simple low cost differentiation of Candida auris from Candida haemulonii complex using CHROMagar Candida medium supplemented with Pal's medium.

    Science.gov (United States)

    Kumar, Anil; Sachu, Arun; Mohan, Karthika; Vinod, Vivek; Dinesh, Kavitha; Karim, Shamsul

    Candida auris is unique due to its multidrug resistance and misidentification as Candida haemulonii by commercial systems. Its correct identification is important to avoid inappropriate treatments. To develop a cheap method for differentiating C. auris from isolates identified as C. haemulonii by VITEK2. Fifteen C. auris isolates, six isolates each of C. haemulonii and Candida duobushaemulonii, and one isolate of Candida haemulonii var. vulnera were tested using CHROMagar Candida medium supplemented with Pal's agar for better differentiation. On CHROMagar Candida medium supplemented with Pal's agar all C. auris strains showed confluent growth of white to cream colored smooth colonies at 37°C and 42°C after 24 and 48h incubation and did not produce pseudohyphae. The isolates of the C. haemulonii complex, on the contrary, showed poor growth of smooth, light-pink colonies at 24h while at 48h the growth was semiconfluent with the production of pseudohyphae. C. haemulonii complex failed to grow at 42°C. We report a rapid and cheap method using CHROMagar Candida medium supplemented with Pal's agar for differentiating C. auris from isolates identified as C. haemulonii by VITEK2. Copyright © 2017 Asociación Española de Micología. Publicado por Elsevier España, S.L.U. All rights reserved.

  6. DietPal: a Web-based dietary menu-generating and management system.

    Science.gov (United States)

    Noah, Shahrul A; Abdullah, Siti Norulhuda; Shahar, Suzana; Abdul-Hamid, Helmi; Khairudin, Nurkahirizan; Yusoff, Mohamed; Ghazali, Rafidah; Mohd-Yusoff, Nooraini; Shafii, Nik Shanita; Abdul-Manaf, Zaharah

    2004-01-30

    Attempts in current health care practice to make health care more accessible, effective, and efficient through the use of information technology could include implementation of computer-based dietary menu generation. While several of such systems already exist, their focus is mainly to assist healthy individuals calculate their calorie intake and to help monitor the selection of menus based upon a prespecified calorie value. Although these prove to be helpful in some ways, they are not suitable for monitoring, planning, and managing patients' dietary needs and requirements. This paper presents a Web-based application that simulates the process of menu suggestions according to a standard practice employed by dietitians. To model the workflow of dietitians and to develop, based on this workflow, a Web-based system for dietary menu generation and management. The system is aimed to be used by dietitians or by medical professionals of health centers in rural areas where there are no designated qualified dietitians. First, a user-needs study was conducted among dietitians in Malaysia. The first survey of 93 dietitians (with 52 responding) was an assessment of information needed for dietary management and evaluation of compliance towards a dietary regime. The second study consisted of ethnographic observation and semi-structured interviews with 14 dietitians in order to identify the workflow of a menu-suggestion process. We subsequently designed and developed a Web-based dietary menu generation and management system called DietPal. DietPal has the capability of automatically calculating the nutrient and calorie intake of each patient based on the dietary recall as well as generating suitable diet and menu plans according to the calorie and nutrient requirement of the patient, calculated from anthropometric measurements. The system also allows reusing stored or predefined menus for other patients with similar health and nutrient requirements. We modeled the workflow of

  7. Cloning and construction of recombinant palI gene from Klebsiella oxytoca on pET-32b into E. coli BL21 (DE3) pLysS for production of isomaltulose, a new generation of sugar

    Energy Technology Data Exchange (ETDEWEB)

    Moeis, Maelita R., E-mail: sony@sith.itb.ac.id; Berlian, Liska, E-mail: sony@sith.itb.ac.id; Suhandono, Sony, E-mail: sony@sith.itb.ac.id; Prima, Alex, E-mail: sony@sith.itb.ac.id; Komalawati, Eli, E-mail: sony@sith.itb.ac.id; Kristianti, Tati, E-mail: sony@sith.itb.ac.id

    2014-03-24

    Klebsiella oxytoca produces sucrose isomerase which catalyses the conversion of sucrose to isomaltulose, a new generation of sugar. From the previous study, palI gene from Klebsiella oxytoca was succesfully isolated from sapodilla fruit (Manilkara zapota). The full-length palI gene sequence of Klebsiella oxytoca was cloned in E. coli DH5α. The deduced amino acid sequence shows 498 residues which includes conserved motif for sucrose isomerisation {sup 325}RLDRD{sup 329} and 97% identical to palI gene from Klebsiella sp. LX3 (GenBank:AAK82938.1). This fragment was succesfullly ligated into the expression vector pET-32b using overlap-extension PCR and cloned in Escherichia coli BL21 (DE3) pLysS. DNA sequencing result shows that palI gene of Klebsiella oxytoca was inserted in-frame in pET-32b. This is the first report on cloning of palI gene from Klebsiella oxytoca.

  8. Engineering of Saccharomyces cerevisiae for production of resveratrol and its derivatives

    DEFF Research Database (Denmark)

    Li, Mingji

    were published in Applied and Environmental Microbiology (Jendresen et al., 2015). I then constructed the TAL pathway to resveratrol by expressing the two TAL genes in combination with 4-coumaryl-CoA ligase (4CL) from Arabidopsis thaliana and resveratrol synthase (VST) from Vitis vinifera in S...... carbon sources. I adopted rational metabolic engineering rational design strategies, synthetic biology techniques and system biology approaches to engineer and analyse yeast cell factories. There are two biosynthesis pathways of resveratrol, starting from tyrosine or phenylalanine, which in this thesis...... are defined as the TAL pathway and PAL pathway respectively. For the TAL pathway, I collaborated with Christian Bille Jendresen, Steen Gustav Stahlhut to screen fourteen diverse heterologous tyrosine ammonia lyases (TALs) for their activity in yeast and E. coli. I expressed the 14 TALs in S. cerevisiae...

  9. Congenital Adrenal Hyperplasia due to 17-alpha-hydoxylase/17,20-lyase Deficiency Presenting with Hypertension and Pseudohermaphroditism: First Case Report from Oman

    Directory of Open Access Journals (Sweden)

    Waad-Allah S. Mula-Abed

    2014-01-01

    Full Text Available This is the first report of congenital adrenal hyperplasia (CAH due to combined 17α-hydroxylase/17,20 lyase deficiency in an Omani patient who was initially treated for many years as a case of hypertension. CAH is an uncommon disorder that results from a defect in steroid hormones biosynthesis in the adrenal cortex. The clinical presentation depends on the site of enzymatic mutations and the types of accumulated steroid precursors. A 22-year-old woman who was diagnosed to have hypertension since the age of 10 years who was treated with anti-hypertensive therapy was referred to the National Diabetes and Endocrine Centre, Royal Hospital, Oman. The patient also had primary amenorrhea and features of sexual infantilism. Full laboratory and radio-imaging investigations were done. Adrenal steroids, pituitary function and karyotyping study were performed and the diagnosis was confirmed by molecular mutation study. Laboratory investigations revealed adrenal steroids and pituitary hormones profile in addition to 46XY karyotype that are consistent with the diagnosis of CAH due to 17α-hydroxylase deficiency. Extensive laboratory workup revealed low levels of serum cortisol (and its precursors 17α-hydroxyprogesterone and 11-deoxycortisol, adrenal androgens (dehydroepiandrosterone sulfate and androstenedione, and estrogen (estradiol; and high levels of mineralocorticoids precursors (11-deoxycorticosterone and corticosterone with high levels of ACTH, FSH and LH. Mutation analysis revealed CYP17A1-homozygous mutation (c.287G>A p.Arg96Gln resulting in the complete absence of 17α-hydroxylase/17,20-lyase activity. The patient was treated with dexamethasone and ethinyl estradiol with cessation of anti-hypertensive therapy. A review of the literature was conducted to identify previous studies related to this subtype of CAH. This is the first biochemically and genetically proven case of CAH due to 17α-hydroxylase/17,20-lyase deficiency in Oman and in the Arab

  10. Regulation of Flavonoid Biosynthetic Genes in Germinating Arabidopsis Seedlings.

    Science.gov (United States)

    Kubasek, WL; Shirley, BW; McKillop, A; Goodman, HM; Briggs, W; Ausubel, FM

    1992-01-01

    Many higher plants, including Arabidopsis, transiently display purple anthocyanin pigments just after seed germination. We observed that steady state levels of mRNAs encoded by four flavonoid biosynthetic genes, PAL1 (encoding phenylalanine ammonia-lyase 1), CHS (encoding chalcone synthase), CHI (encoding chalcone isomerase), and DFR (encoding dihydroflavonol reductase), were temporally regulated, peaking in 3-day-old seedlings grown in continuous white light. Except for the case of PAL1 mRNA, mRNA levels for these flavonoid genes were very low in seedlings grown in darkness. Light induction studies using seedlings grown in darkness showed that PAL1 mRNA began to accumulate before CHS and CHI mRNAs, which, in turn, began to accumulate before DFR mRNA. This order of induction is the same as the order of the biosynthetic steps in flavonoid biosynthesis. Our results suggest that the flavonoid biosynthetic pathway is coordinately regulated by a developmental timing mechanism during germination. Blue light and UVB light induction experiments using red light- and dark-grown seedlings showed that the flavonoid biosynthetic genes are induced most effectively by UVB light and that blue light induction is mediated by a specific blue light receptor. PMID:12297632

  11. Elimination of hydrogen sulphide and β substitution in cystein, catalyzed by the cysteine-lyase of hens yolk-sac and yolk (1961)

    International Nuclear Information System (INIS)

    Chapeville, F.; Fromageot, P.

    1961-01-01

    The yolk of incubated hen's eggs contains a pyridoxal phosphate activated enzyme, free of iron, copper, magnesium and calcium. This enzyme activates the β-carbon atom of cysteine. Its reactivity is demonstrated by the ease with which this β-carbon fixes various sulfur containing substances in which the sulfur has reducing properties: inorganic sulfide, sulfide or cysteine itself. In the absence of substances able to react with the β-carbon atom, the active complex, consisting of the enzyme and the aminated tri-carbon chain, is hydrolysed to pyruvic acid and ammonia. The liberation of hydrogen sulfide thus appears to be the consequence either of the substitution of the β-carbon atom of cysteine or of the decomposition of the complex which this aminoacid forms with the enzyme studied. The latter seems therefore to possess an activity which differs from the activity of the desulfhydrases as yet known. We suggest to call this enzyme cystein-lyase. (authors) [fr

  12. The anaerobic chytridiomycete fungus Piromyces sp. E2 produces ethanol via pyruvate:formate lyase and an alcohol dehydrogenase E.

    NARCIS (Netherlands)

    Boxma, B.; Voncken, F.L.M.; Jannink, S.A.; Alen, T.A. van; Akhmanova, A.S.; Weelden, S.W. van; Hellemond, J.J. van; Ricard, G.N.S.; Huynen, M.A.; Tielens, A.G.; Hackstein, J.H.P.

    2004-01-01

    Anaerobic chytridiomycete fungi possess hydrogenosomes, which generate hydrogen and ATP, but also acetate and formate as end-products of a prokaryotic-type mixed-acid fermentation. Notably, the anaerobic chytrids Piromyces and Neocallimastix use pyruvate:formate lyase (PFL) for the catabolism of

  13. Zur Geschichte der Geowissenschaften im Museum für Naturkunde zu Berlin. Teil 6: Geschichte des Geologisch-Paläontologischen Instituts und Museums der Universität Berlin 1910–2004

    Directory of Open Access Journals (Sweden)

    W. Gross

    2004-01-01

    Full Text Available Die Entwicklung des Geologisch-Paläontologischen Instituts und Museums der Universität Berlin von einer Institution, die Geologie zusammen mit Paläontologie als eine Einheit vertrat, über eine Institution, die eine geotektonische Ausrichtung hatte, zu einer auf Paläontologie konzentrierten Institution wird nachvollzogen. Die beiden Institutsdirektoren am Anfang des 20sten Jahrhunderts waren Vertreter der allumfassenden Geologie des 19ten Jahrhunderts, während die beiden folgenden Direktoren eine Geologie ohne Paläontologie vertraten. Das führte zu einer Trennung der beiden Richtungen, und nach der III. Hochschulreform der DDR 1968 verblieb allein die sammlungsbezogene Paläontologie am Museum. Nach der Wiedervereinigung wurde ein Institut für Paläontologie mit biologischer Ausrichtung mit zwei Professuren, einer für Paläozoologie und einer für Paläobotanik, eingerichtet. The development of the Geologisch-Paläontologisches Institut und Museum of the Museum für Naturkunde at the Humboldt University (formerly Friedrich-Wilhelm-Universität in Berlin from a geology-paleontology institution to a pure paleontology institution is described. The first two directors of the department in the beginning of the 20th century, Prof, von Branca and Prof. Pompeckj, represented a 19th century concept of a geology, which included paleontology, even vertebrate paleontology as the crown jewel of geology. They fought sometimes vigorously against a separation of paleontology from geology. The next two directors. Prof. Stille and Prof, von Bubnoff, were the leading geologists in Germany; to be a student of Stille was a special trade mark in geology of Germany. They represented a geology centered on tectonics. The separation of paleontology as separate section was prepared. The destructions of the Second World War, the following restaurations and the division of Germany into two States influenced strongly their directorships. The education of geologists

  14. Identification, expression, and characterization of a novel bacterial RGI Lyase enzyme for the production of bio-functional fibers

    DEFF Research Database (Denmark)

    da Silva, Ines Isabel Cardoso Rodrigues; Larsen, Dorte Møller; Meyer, Anne S.

    2011-01-01

    A gene encoding a putative rhamnogalacturonan I (RGI) Lyase (EC 4.2.2.-) from Bacillus licheniformis (DSM13) was selected after a homology search and phylogenetic analysis and optimized with respect to codon usage. The designed gene was transformed into Pichia pastoris and the enzyme was produced...

  15. Involvement of plasma membrane peroxidases and oxylipin pathway in the recovery from phytoplasma disease in apple (Malus domestica).

    Science.gov (United States)

    Patui, Sonia; Bertolini, Alberto; Clincon, Luisa; Ermacora, Paolo; Braidot, Enrico; Vianello, Angelo; Zancani, Marco

    2013-06-01

    Apple trees (Malus domestica Borkh.) may be affected by apple proliferation (AP), caused by 'Candidatus Phytoplasma mali'. Some plants can spontaneously recover from the disease, which implies the disappearance of symptoms through a phenomenon known as recovery. In this article it is shown that NAD(P)H peroxidases of leaf plasma membrane-enriched fractions exhibited a higher activity in samples from both AP-diseased and recovered plants. In addition, an increase in endogenous SA was characteristic of the symptomatic plants, since its content increased in samples obtained from diseased apple trees. In agreement, phenylalanine ammonia lyase (PAL) activity, a key enzyme of the phenylpropanoid pathway, was increased too. Jasmonic acid (JA) increased only during recovery, in a phase subsequent to the pathological state, and in concomitance to a decline of salicylic acid (SA). Oxylipin pathway, responsible for JA synthesis, was not induced during the development of AP-disease, but it appeared to be stimulated when the recovery occurred. Accordingly, lipoxygenase (LOX) activity, detected in plasma membrane-enriched fractions, showed an increase in apple leaves obtained from recovered plants. This enhancement was paralleled by an increase of hydroperoxide lyase (HPL) activity, detected in leaf microsomes, albeit the latter enzyme was activated in either the disease or recovery conditions. Hence, a reciprocal antagonism between SA- and JA-pathways could be suggested as an effective mechanism by which apple plants react to phytoplasma invasions, thereby providing a suitable defense response leading to the establishment of the recovery phenomenon. Copyright © Physiologia Plantarum 2012.

  16. Health-related factors correlate with behavior trends in physical activity level in old age: longitudinal results from a population in São Paulo, Brazil

    Directory of Open Access Journals (Sweden)

    Ribeiro Manoel CSA

    2010-11-01

    Full Text Available Abstract Background Physical inactivity in leisure time is common among elderly in Brazil and this finding is particularly alarming considering that this population is greatly affected by chronic diseases. The identification of health factors that influence physical activity level (PAL will help in the development of strategies for increasing PAL older adults. The current research aimed to identify variables that independently affect behavior trends in PAL over the course of two years among elderly. Methods A survey entitled the Epidoso Project ("Epidemiology of aging" studied 1,667 community-based older individuals in São Paulo city, Brazil over the course of two years. Physical activity level was determined through questions about frequency and duration of physical activities. Body Mass Index was calculated; functional capacity was assessed through the ADL (activities of daily living scale; cognition was assessed by Mini-Mental State Examination; and mental health was assessed through the Dysthymia Screening. Experiences of falls and fractures were also assessed. Subjects were divided into three groups according to their self-report of Physical Activity Level: a - Regularly Active; b - Insufficiently Active and c - Physically Inactive. Behavior trends in PAL were also measured after two years. Multivariate regression model methodology was used to test associations longitudinally. Results Results from the final model demonstrated that the risk of a not favorable behavior trend in PAL, which included the group who remained physically inactive and the group that displayed decreased PAL, in this cohort of older adults was significantly increased if the individual was female (OR = 2.50; 95% CI = 1.60-3.89; P , older (80 y vs. 65 y, OR = 6.29, 95% CI = 2.69-14.67; P , dependent on help from others for activities in the ADL scale (moderate-severe = 4-7+ vs. 0 ADLs (OR = 2.25, 95% CI = 1.20-4.21; P or had experienced a history of falls with

  17. Five phosphonate operon gene products as components of a multi-subunit complex of the carbon-phosphorus lyase pathway

    DEFF Research Database (Denmark)

    Jochimsen, Bjarne; Lolle, Signe; McSorley, Fern R.

    2011-01-01

    Organophosphonate utilization by Escherichia coli requires the 14 cistrons of the phnCDEFGHIJKLMNOP operon, of which the carbon-phosphorus lyase has been postulated to consist of the seven polypeptides specified by phnG to phnM. A 5,660-bp DNA fragment encompassing phnGHIJKLM is cloned, followed...

  18. The pectin lyase-encoding gene (pnl) family from Glomerella cingulata: characterization of pnlA and its expression in yeast.

    Science.gov (United States)

    Templeton, M D; Sharrock, K R; Bowen, J K; Crowhurst, R N; Rikkerink, E H

    1994-05-03

    Oligodeoxyribonucleotide primers were designed from conserved amino acid (aa) sequences between pectin lyase D (PNLD) from Aspergillus niger and pectate lyases A and E (PELA/E) from Erwinia chrysanthemi. The polymerase chain reaction (PCR) was used with these primers to amplify genomic DNA from the plant pathogenic fungus Glomerella cingulata. Three different 220-bp fragments with homology to PNL-encoding genes from A. niger, and a 320-bp fragment with homology to PEL-encoding genes from Nicotiana tabacum and E. carotovora were cloned. One of the 220-bp PCR products (designated pnlA) was used as a probe to isolate a PNL-encoding gene from a lambda genomic DNA library prepared from G. cingulata. Nucleotide (nt) sequence data revealed that this gene has seven exons and codes for a putative 380-aa protein. The nt sequence of a cDNA clone, prepared using PCR, confirmed the presence of the six introns. The positions of the introns were different from the sites of the five introns present in the three PNL-encoding genes previously sequenced from A. niger. PNLA was synthesised in yeast by cloning the cDNA into the expression vector, pEMBLYex-4, and enzymatically active protein was secreted into the culture medium. Significantly higher expression was achieved when the context of the start codon, CACCATG, was mutated to CAAAATG, a consensus sequence commonly found in highly expressed yeast genes. The produced protein had an isoelectric point (pI) of 9.4, the same as that for the G. cingulata pnlA product.(ABSTRACT TRUNCATED AT 250 WORDS)

  19. Longitudinal Relationship Among Physical Fitness, Walking-Related Physical Activity, and Fatigue in Children With Cerebral Palsy

    NARCIS (Netherlands)

    Balemans, A.C.J.; van Wely, L.; Becher, J.G.; Dallmeijer, A.J.

    2015-01-01

    Background. A vicious circle of decreased physical fitness, early fatigue, and low physical activity levels (PAL) is thought to affect children with cerebral palsy (CP). However, the relationship of changes in physical fitness to changes in PAL and fatigue is unclear. Objective. The objective of

  20. Gene expression in the lignin biosynthesis pathway during soybean seed development.

    Science.gov (United States)

    Baldoni, A; Von Pinho, E V R; Fernandes, J S; Abreu, V M; Carvalho, M L M

    2013-02-28

    The study of gene expression in plants is fundamental, and understanding the molecular mechanisms involved in important biological processes, such as biochemical pathways or signaling that are used or manipulated in improvement programs, are key for the production of high-quality soybean seeds. Reports related to gene expression of lignin in seeds are scarce in the literature. We studied the expression of the phenylalanine ammonia-lyase (PAL), cinnamate 4-hydroxylase, 4-hydroxycinnamate 3-hydroxylase, and cinnamyl alcohol dehydrogenase genes involved in lignin biosynthesis during the development of soybean (Glycine max L. Merrill) seeds. As the endogenous control, the eukaryotic elongation factor 1-beta gene was used in two biological replicates performed in triplicate. Relative quantitative expression of these genes during the R4, R5, R6, and R7 development stages was analyzed. Real-time polymerase chain reaction was used for the gene expression study. The analyses were carried out in an ABI PRISM 7500 thermocycler using the comparative Ct method and SYBR Green to detect amplification. The seed samples at the R4 stage were chosen as calibrators. Increased expression of the cinnamate-4-hydroxylase and PAL genes occurred in soybean seeds at the R5 and R6 development stages. The cinnamyl alcohol dehydrogenase gene was expressed during the final development phases of soybean seeds. In low-lignin soybean cultivars, the higher expression of the PAL gene occurs at development stages R6 and R7. Activation of the genes involved in the lignin biosynthesis pathway occurs at the beginning of soybean seed development.

  1. Ammonium-related metabolic changes affect somatic embryogenesis in pumpkin (Cucurbita pepo L.).

    Science.gov (United States)

    Mihaljević, Snježana; Radić, Sandra; Bauer, Nataša; Garić, Rade; Mihaljević, Branka; Horvat, Gordana; Leljak-Levanić, Dunja; Jelaska, Sibila

    2011-11-01

    Somatic embryogenesis in pumpkin can be induced on auxin-containing medium and also on hormone-free medium containing 1mM ammonium (NH(4)(+)) as the sole source of nitrogen. Growth of NH(4)(+)-induced embryogenic tissue was slow and caused considerable acidification of the culture medium. Small spherical cells with dense cytoplasma formed proembryogenic cell clusters that could not develop into late stage embryos. Buffering of NH(4)(+) medium with 25mM 2-(N-morpholino)-ethane-sulfonic acid enhanced tissue proliferation, but no further differentiation was observed. Later stage embryos developed only after re-supply of nitrogen in form of nitrate or l-glutamine. Effects of nitrogen status and pH of culture media on ammonium assimilation were analyzed by following the activity of glutamine synthetase (GS) in relation to phenylalanine ammonia-lyase (PAL). Increased activity of GS and PAL in NH(4)(+) induced tissue coincided with significantly higher activity of stress-related enzymes superoxide dismutase (SOD) and soluble peroxidase (POD), indicating oxidative stress response of embryogenic tissue to NH(4)(+) as the sole source of nitrogen. In addition, considerable increase was observed in callose accumulation and esterase activity, the early markers of somatic embryogenesis. Activity of stress-related enzymes decreased after the re-supply of nitrate (20mM) or Gln (10mM) in combination with NH(4)(+) (1mM), which subsequently triggered globular embryo development. Together, these results suggest that stress responses, as affected by nitrogen supply, contribute to the regulation of embryogenic competence in pumpkin. Copyright © 2011 Elsevier GmbH. All rights reserved.

  2. Prunus domestica pathogenesis-related protein-5 activates the defense response pathway and enhances the resistance to fungal infection.

    Directory of Open Access Journals (Sweden)

    Ashraf El-kereamy

    Full Text Available Pathogenesis-related protein-5 (PR-5 has been implicated in plant disease resistance and its antifungal activity has been demonstrated in some fruit species. However, their roles, especially their interactions with the other defense responses in plant cells, are still not fully understood. In this study, we have cloned and characterized a new PR-5 cDNA named PdPR5-1 from the European plum (Prunus domestica. Expression of PdPR5-1 was studied in different cultivars varying in resistance to the brown rot disease caused by the necrotrophic fungus Monilinia fructicola. In addition transgenic Arabidopsis, ectopically expressing PdPR5-1 was used to study its role in other plant defense responses after fungal infection. We show that the resistant cultivars exhibited much higher levels of transcripts than the susceptible cultivars during fruit ripening. However, significant rise in the transcript levels after infection with M. fructicola was observed in the susceptible cultivars too. Transgenic Arabidopsis plants exhibited more resistance to Alternaria brassicicola. Further, there was a significant increase in the transcripts of genes involved in the phenylpropanoid biosynthesis pathway such as phenylalanine ammonia-lyase (PAL and phytoalexin (camalexin pathway leading to an increase in camalexin content after fungal infection. Our results show that PdPR5-1 gene, in addition to its anti-fungal properties, has a possible role in activating other defense pathways, including phytoalexin production.

  3. Evidence for some signal transduction elements involved in UV-light-dependent responses in parsley protoplasts

    International Nuclear Information System (INIS)

    Frohnmeyer, H.; Bowler, C.; Schäfer, E.

    1997-01-01

    The signalling pathways used by UV-light are largely unknown. Using protoplasts from a heterotrophic parsley (Petroselinum crispum L.) cell culture that exclusively respond to UV-B light between 300 and 350 nm with a fast induction of genes encoding flavonoid biosynthetic enzymes, information was obtained about the UV-light signal transduction pathway for chalcone synthase (CHS) and phenylalanine ammonia-lyase (PAL) gene expression. Pharmacological effectors which influence intracellular calcium levels, calmodulin and the activity of serine/threonine kinases also changed the UV-light-dependent expression of these genes. This evaluation indicated the participation of these components on the UV-B-mediated signal transduction cascade to CHS. In contrast, neither membrane-permeable cyclic GMP nor the tyrosine kinase inhibitor genistein affected CHS or PAL expression. Similar results were obtained in protoplasts, which have been transiently transformed with CHS-promoter/GUS (β-glucuronidase) reporter fusion constructs. The involvement of calcium and calmodulin was further indicated in a cell-free light-responsive in vitro transcription system from evacuolated parsley protoplasts. In conclusion, there is evidence now that components of the UV-light-dependent pathway leading to the CHS-promoter are different from the previously characterized cGMP-dependent pathway to CHS utilized by phytochrome in soybean (Glycine max) and tomato seedlings (Lycopersicon esculentum). (author)

  4. Chitosan and grape secondary metabolites: A proteomics and metabolomics approach

    Directory of Open Access Journals (Sweden)

    Bavaresco Luigi

    2017-01-01

    Full Text Available Chitosan is a polysaccharide obtained by deacetylation of chitin, and it is involved in defence mechanisms of plants toward diseases. In the present work, V. vinifera L. cv. Ortrugo, grafted on 420A rootstock was grown in pot and treated, at veraison, by 0.03% chitosan solution at cluster level. Just before the treatment (T0 and 24 hours (T1, 48 hours (T2, 72 hours (T3 and 10 days (T4 later, the concentration of stilbenic compounds was detected, and at T1 proteomics and metabolomics analyses were done. Proteomics relies on the analysis of the complete set of proteins existing in a given substrate, while metabolomics relies on the analyses of the complete set of metabolites in a given substrate. The treatment improved the stilbene concentration over the control at T1. Proteomic analysis showed that superoxide dismutase (SOD and phenylalanine ammonia-lyase (PAL were overexpressed in the treated grapes. SOD is known to be an enzyme active against reactive oxygen species (ROS while PAL is a key enzyme in the phenylpropanoids pathway. Metabolomics analysis highlighted the positive role of the treatment in improving the triperpenoid concentration (betulin, erythrodiol, uvaol, oleanolate; these compounds are known to be effective against microbes, insects and fungi.

  5. Identification of transcription factors ZmMYB111and ZmMYB148 involved in phenylpropanoid metabolism

    Directory of Open Access Journals (Sweden)

    Junjie eZhang

    2016-02-01

    Full Text Available Maize is the leading crop worldwide in terms of both planting area and total yields, but environmental stresses cause significant losses in productivity. Phenylpropanoid compounds play an important role in plant stress resistance; however, the mechanism of their synthesis is not fully understood, especially in regard to the expression and regulation of key genes. Phenylalanine ammonia-lyase (PAL is the first key enzyme involved in phenylpropanoid metabolism, and it has a significant effect on the synthesis of important phenylpropanoid compounds. According to the results of sequence alignments and functional prediction, we selected two conserved R2R3-MYB transcription factors as candidate genes for the regulation of phenylpropanoid metabolism. The two candidate R2R3-MYB genes, which we named ZmMYB111and ZmMYB148, were cloned, and then their structural characteristics and phylogenetic placement were predicted and analyzed. In addition, a series of evaluations were performed, including expression profiles, subcellular localization, transcription activation, protein-DNA interaction, and transient expression in maize endosperm. Our results indicated that both ZmMYB111 and ZmMYB148 are indeed R2R3-MYB transcription factors and that they may play a regulatory role in PAL gene expression.

  6. Growth and phenolic compounds of Lactuca sativa L. grown in a closed-type plant production system with UV-A, -B, or -C lamp.

    Science.gov (United States)

    Lee, Min-Jeong; Son, Jung Eek; Oh, Myung-Min

    2014-01-30

    The production of high-quality crops based on phytochemicals is a strategy for accelerating the practical use of plant factories. Previous studies have demonstrated that ultraviolet (UV) light is effective in improving phytochemical production. This study aimed to determine the effect of various UV wavelengths on growth and phenolic compound accumulation in lettuce (Lactuca sativa L.) grown in a closed-type plant production system. Seven days, 1 day and 0.25 day were determined as the upper limit of the irradiation periods for UV-A, -B, and -C, respectively, in the lettuce based on physiological disorders and the fluorescence parameter F(v)/F(m). Continuous UV-A treatment significantly induced the accumulation of phenolic compounds and antioxidants until 4 days of treatment without growth inhibition, consistent with an increase in phenylalanine ammonia lyase (PAL) gene expression and PAL activity. Repeated or gradual UV-B exposure yielded approximately 1.4-3.6 times more total phenolics and antioxidants, respectively, than the controls did 2 days after the treatments, although both treatments inhibited lettuce growth. Repeated UV-C exposure increased phenolics but severely inhibited the growth of lettuce plants. Our data suggest that UV irradiation can improve the accumulation of phenolic compounds with antioxidant properties in lettuce cultivated in plant factories. © 2013 Society of Chemical Industry.

  7. Differential metabolism of L-phenylalanine in the formation of aromatic volatiles in melon (Cucumis melo L.) fruit.

    Science.gov (United States)

    Gonda, Itay; Davidovich-Rikanati, Rachel; Bar, Einat; Lev, Shery; Jhirad, Pliaa; Meshulam, Yuval; Wissotsky, Guy; Portnoy, Vitaly; Burger, Joseph; Schaffer, Arthur A; Tadmor, Yaakov; Giovannoni, James J; Fei, Zhangjun; Fait, Aaron; Katzir, Nurit; Lewinsohn, Efraim

    2018-04-01

    Studies on the active pathways and the genes involved in the biosynthesis of L-phenylalanine-derived volatiles in fleshy fruits are sparse. Melon fruit rinds converted stable-isotope labeled L-phe into more than 20 volatiles. Phenylpropanes, phenylpropenes and benzenoids are apparently produced via the well-known phenylpropanoid pathway involving phenylalanine ammonia lyase (PAL) and being (E)-cinnamic acid a key intermediate. Phenethyl derivatives seemed to be derived from L-phe via a separate biosynthetic route not involving (E)-cinnamic acid and PAL. To explore for a biosynthetic route to (E)-cinnamaldehyde in melon rinds, soluble protein cell-free extracts were assayed with (E)-cinnamic acid, CoA, ATP, NADPH and MgSO 4 , producing (E)-cinnamaldehyde in vitro. In this context, we characterized CmCNL, a gene encoding for (E)-cinnamic acid:coenzyme A ligase, inferred to be involved in the biosynthesis of (E)-cinnamaldehyde. Additionally we describe CmBAMT, a SABATH gene family member encoding a benzoic acid:S-adenosyl-L-methionine carboxyl methyltransferase having a role in the accumulation of methyl benzoate. Our approach leads to a more comprehensive understanding of L-phe metabolism into aromatic volatiles in melon fruit. Copyright © 2018 Elsevier Ltd. All rights reserved.

  8. Physical activity level and exercise in patients with diabetes mellitus.

    Science.gov (United States)

    Duarte, Camila Kümmel; Almeida, Jussara Carnevale de; Merker, Aline Juliana Schneider; Brauer, Fabiane de Oliveira; Rodrigues, Ticiana da Costa

    2012-01-01

    To compare physical activity level (PAL) and care related to exercise in patients with diabetes mellitus (DM). DM outpatients (adult, insulin-user patients) were assessed for PAL (international questionnaire; moderate- and high-level activities, as well as walking, over a typical week) and questioned about formal exercise practice, self-care, and hypoglycemic episodes related to exercise or reasons for not exercising. Two hundred twenty five patients were assessed: 107 (47.6%) had type 2 diabetes mellitus (DM2) and 118 (52.4%) had type 1 diabetes mellitus (DM1), with a larger percentage of patients with DM2 being classified as poorly active [33 (30.7%) versus 12 (10.3%)] and a lower percentage being classified as highly active [9 (8.7%) versus 29 (25%)], compared with patients having DM1. Patients who do not exercise (n = 140) gave different reasons for not doing so: patients with DM2 claimed that they "felt uncomfortable", "presented medical restrictions", and "did not like it"; DM1 patients claimed that they "had no time to exercise", "were lazy", and "had hypoglycemic episodes". Only 85 patients exercised regularly, regardless of the PAL, and 38.8% performed self-care, such as eating, stretching, and capillary glucose monitoring. Patients with DM2 [5 (14.3%)] reported a lower number of hypoglycemic episodes related to exercise than those with DM1 [17 (34%)]. Patients with DM2 have different PAL and behavior related to exercise than those seen in DM1 patients.

  9. Pengaruh Karakteristik Budaya Organisasi, Locus of Control dan Disiplin Kerja Terhadap Kinerja Karyawan Studi Pada Perusahaan Daerah Pengelola Air Limbah (PD PAL Kota Banjarmasin.

    Directory of Open Access Journals (Sweden)

    Dessy Rosiana

    2016-09-01

    Full Text Available This research was aimed to simultaneously analyze the significantly positive effect of organizational culture characteristics, locus of control and work discipline towards employee performance at PD PAL Banjarmasin; also to partially analyze the significantly positive effect of organizational culture characteristics, locus of control, and work discipline towards employee performance at PD PAL Banjarmasin. The type of this research was explanatory research. All population became the samples in this research. The information collection of 53 employees who acted as the respondents in this research was done by using questionnaire arranged in 5 Likert Scale. The data analysis used multiple linear regression analysis, multiple determination coefficient, as well as F test and t test. The result of this research showed that the variable of organizational culture characteristics, locus of control and work discipline simultaneously has a significantly positive effect towards employee performance at PD PAL Banjarmasin. The organizational culture characteristics partially does not  have a positive and significant effect towards employee performance. Partially   the locus control has a significantly positive effect towards employee performance. Work discipline partially has a significantly positive effect towards employee performance. The implication to the management, based on the result of this research, is (1 the implementation of the organizational culture characteristics needs to be socialized among employee of PD PAL, and (2 a strong control on the locus control and discipline is needed to improve the employee performance. Keywords: Organizational Culture Characteristics, Locus of Control, Work Discipline, Employee Performance

  10. Estimation of physical activity levels using cell phone questionnaires: a comparison with accelerometry for evaluation of between-subject and within-subject variations.

    Science.gov (United States)

    Bexelius, Christin; Sandin, Sven; Trolle Lagerros, Ylva; Litton, Jan-Eric; Löf, Marie

    2011-09-25

    Physical activity promotes health and longevity. Further elaboration of the role of physical activity for human health in epidemiological studies on large samples requires accurate methods that are easy to use, cheap, and possible to repeat. The use of telecommunication technologies such as cell phones is highly interesting in this respect. In an earlier report, we showed that physical activity level (PAL) assessed using a cell phone procedure agreed well with corresponding estimates obtained using the doubly labeled water method. However, our earlier study indicated high within-subject variation in relation to between-subject variations in PAL using cell phones, but we could not assess if this was a true variation of PAL or an artifact of the cell phone technique. Our objective was to compare within- and between-subject variations in PAL by means of cell phones with corresponding estimates using an accelerometer. In addition, we compared the agreement of daily PAL values obtained using the cell phone questionnaire with corresponding data obtained using an accelerometer. PAL was measured both with the cell phone questionnaire and with a triaxial accelerometer daily during a 2-week study period in 21 healthy Swedish women (20 to 45 years of age and BMI from 17.7 kg/m² to 33.6 kg/m²). The results were evaluated by fitting linear mixed effect models and descriptive statistics and graphs. With the accelerometer, 57% (95% confidence interval [CI] 40%-66%) of the variation was within subjects, while with the cell phone, within-subject variation was 76% (95% CI 59%-83%). The day-to-day variations in PAL observed using the cell phone questions agreed well with the corresponding accelerometer results. Both the cell phone questionnaire and the accelerometer showed high within-subject variations. Furthermore, day-to-day variations in PAL within subjects assessed using the cell phone agreed well with corresponding accelerometer values. Consequently, our cell phone

  11. The ketogenic diet is well tolerated and can be effective in patients with argininosuccinate lyase deficiency and refractory epilepsy

    NARCIS (Netherlands)

    Peuscher, Rosanne; Dijsselhof, Monique E.; Abeling, Nico G.; van Rijn, Margreet; van Spronsen, Francjan J.; Bosch, Annet M.

    2012-01-01

    Argininosuccinate lyase (ASL) deficiency (MIM 608310, McKusick 207900) is a rare disorder of the urea cycle, which leads to a deficiency of arginine and hyperammonemia. Epilepsy is a frequent complication of this disorder. A ketogenic diet (KD) can be a very effective therapy for refractory

  12. Measurements of Transverse Emittance for RF Photocathode Gun at the PAL

    CERN Document Server

    Park Jang Ho; Park, Sung-Ju; Soo Ko In; Wang, Xijie; Woon Parc, Yong; Xiang, Dao

    2005-01-01

    A BNL GUN-IV type RF photo-cathode gun is under fabrication for use in the FIR (Far Infra-Red) facility being built at the Pohang Accelerator Laboratory (PAL). Performance test of the gun will include the measurement of transverse emittance profile along the longitudinal direction. Successful measurement of the emittance profile will provide powerful tool for the commissioning of the 4GLS (4th generation light source) injectors based on the emittance compensation principle. We are going to achieve this withthe use of pepper-pot based emittance meters that can be moved along the longitudinal direction. In this article, we present design considerations on the emittance meter with the resolution of 1 mm mrad.

  13. The Salmonella effector protein SpvC, a phosphothreonine lyase is functional in plant cells

    Directory of Open Access Journals (Sweden)

    Christina eNeumann

    2014-10-01

    Full Text Available Salmonella is one of the most prominent causes of food poisoning and growing evidence indicates that contaminated fruits and vegetables are an increasing concern for human health. Successful infection demands the suppression of the host immune system, which is often achieved via injection of bacterial effector proteins into host cells. In this report we present the function of Salmonella effector protein in plant cell, supporting the new concept of trans-kingdom competence of this bacterium. We screened a range of Salmonella Typhimurium effector proteins for interference with plant immunity. Among these, the phosphothreonine lyase SpvC attenuated the induction of immunity-related genes when present in plant cells. Using in vitro and in vivo systems we show that this effector protein interacts with and dephosphorylates activated Arabidopsis Mitogen-activated Protein Kinase 6 (MPK6, thereby inhibiting defense signaling. Moreover, the requirement of Salmonella SpvC was shown by the decreased proliferation of the ΔspvC mutant in Arabidopsis plants. These results suggest that some Salmonella effector proteins could have a conserved function during proliferation in different hosts. The fact that Salmonella and other Enterobacteriaceae use plants as hosts strongly suggests that plants represent a much larger reservoir for animal pathogens than so far estimated.

  14. The Salmonella effector protein SpvC, a phosphothreonine lyase is functional in plant cells

    KAUST Repository

    Neumann, Christina

    2014-10-17

    Salmonella is one of the most prominent causes of food poisoning and growing evidence indicates that contaminated fruits and vegetables are an increasing concern for human health. Successful infection demands the suppression of the host immune system, which is often achieved via injection of bacterial effector proteins into host cells. In this report we present the function of Salmonella effector protein in plant cell, supporting the new concept of trans-kingdom competence of this bacterium. We screened a range of Salmonella Typhimurium effector proteins for interference with plant immunity. Among these, the phosphothreonine lyase SpvC attenuated the induction of immunity-related genes when present in plant cells. Using in vitro and in vivo systems we show that this effector protein interacts with and dephosphorylates activated Arabidopsis Mitogen-activated Protein Kinase 6 (MPK6), thereby inhibiting defense signaling. Moreover, the requirement of Salmonella SpvC was shown by the decreased proliferation of the ΔspvC mutant in Arabidopsis plants. These results suggest that some Salmonella effector proteins could have a conserved function during proliferation in different hosts. The fact that Salmonella and other Enterobacteriaceae use plants as hosts strongly suggests that plants represent a much larger reservoir for animal pathogens than so far estimated.

  15. Naringenin inhibits the growth and stimulates the lignification of soybean root

    Directory of Open Access Journals (Sweden)

    Graciene de Souza Bido

    2010-06-01

    Full Text Available The flavanone naringenin, an intermediate in flavonoid biosynthesis, was tested for its effect on root growth, phenylalanine ammonia-lyase (PAL and peroxidase (POD activities, as well as phenolic compounds and lignin contents in soybean (Glycine max L. Merrill seedlings. Three-day-old seedlings were cultivated in half-strength Hoagland nutrient solution (pH 6.0, with or without 0.1 to 0.4 mM naringenin in a growth chamber (25°C, 12-h photoperiod, irradiance of 280 µmol m-2 s-1 for 24 h. Inhibitory effects on root growth (length, weight, cell viability, PAL and soluble POD activities were detected after naringenin treatments. These effects were associated with stimulatory activity of the cell wall-bound POD followed by an increase in the lignin contents, suggesting that naringenin-induced inhibition in soybean roots could be due to the lignification process.Os efeitos de naringenina, um intermediário da biossíntese de flavonóides, foram avaliados sobre o crescimento das raízes, as atividades da fenilalanina amônia liase (PAL e peroxidases, bem como sobre os teores de compostos fenólicos e de lignina em plântulas de soja (Glycine max L. Merrill. Plântulas de três dias foram cultivadas em solução nutritiva de Hoagland, meia-força (pH 6,0, contendo ou não, naringenina 0,1 a 0,4 mM, em uma câmara de germinação (25°C, fotoperíodo de 12 h, 280 µmol m-2 s-1 durante 24 h. Efeitos inibitórios no crescimento das raízes (comprimento, massa e viabilidade celular e nas atividades da PAL e POD solúvel foram constatados após os tratamentos com naringenina. Estes efeitos foram associados com atividade estimulatória da POD ligada à parede celular, seguido por aumento nos teores de lignina, sugerindo que a inibição do crescimento das raízes pode ser devido ao processo de lignificação.

  16. Positron annihilation lifetime spectroscopy (PALS) application in metal barrier layer integrity for porous low- k materials

    CERN Document Server

    Simon, Lin; Gidley, D W; Wetzel, J T; Monnig, K A; Ryan, E T; Simon, Jang; Douglas, Yu; Liang, M S; En, W G; Jones, E C; Sturm, J C; Chan, M J; Tiwari, S C; Hirose, M

    2002-01-01

    Positron Annihilation Lifetime Spectroscopy (PALS) is a useful tool to pre-screen metal barrier integrity for Si-based porous low-k dielectrics. Pore size of low-k, thickness of metal barrier Ta, positronium (Ps) leakage from PALS, trench sidewall morphology, electrical test from one level metal (1LM) pattern wafer and Cu diffusion analysis were all correlated. Macro-porous low-k (pore size >=200 AA) and large scale meso-porous low-k (>50~200 AA) encounter both Ps leakage and Cu diffusion into low-k dielectric in the 0.25 mu mL/0.3 mu mS structures when using SEMATECH in-house PVD Ta 250 AA as barrier layer. For small scale meso-porous (>20~50 AA) and micro- porous (<=20 AA) low-k, no Ps leakage and no Cu diffusion into low-k were observed even with PVD Ta 50 AA, which is proved also owing to sidewall densification to seal all sidewall pores due to plasma etch and ash. For future technology, smaller pore size of porous Si-based low-k (=<50 AA) will be preferential for dense low-k like trench sidewall to...

  17. Crystal Structure of α-1,4-Glucan Lyase, a Unique Glycoside Hydrolase Family Member with a Novel Catalytic Mechanism

    NARCIS (Netherlands)

    Rozeboom, Henriëtte J.; Yu, Shukun; Madrid, Susan; Kalk, Kor H.; Zhang, Ran; Dijkstra, Bauke W.

    2013-01-01

    α-1,4-Glucan lyase (EC 4.2.2.13) from the red seaweed Gracilariopsis lemaneiformis cleaves α-1,4-glucosidic linkages in glycogen, starch, and malto-oligosaccharides, yielding the keto-monosaccharide 1,5-anhydro-D-fructose. The enzyme belongs to glycoside hydrolase family 31 (GH31) but degrades

  18. Fast and slow plasma components produced by the PALS facility - comparsion of interferometric and ion diagnostic measurements

    Czech Academy of Sciences Publication Activity Database

    Pisarcyk, T.; Badziak, J.; Kasperczuk, A.; Parys, P.; Wolowski, J.; Woryna, E.; Jungwirth, Karel; Králiková, Božena; Krása, Josef; Láska, Leoš; Mašek, Karel; Pfeifer, Miroslav; Rohlena, Karel; Skála, Jiří; Ullschmied, Jiří

    2002-01-01

    Roč. 52, Suppl. D (2002), D310-D316 ISSN 0011-4626. [Plasma Physics and Technology. Prague, 10.06.2002-13.06.2002] Grant - others:PALS/005(6)(XE) HPRI-CT-1999-00053 Institutional research plan: CEZ:AV0Z1010921 Keywords : laser plasma Subject RIV: BL - Plasma and Gas Discharge Physics Impact factor: 0.311, year: 2002

  19. Physiochemical and Phytochemical Properties of Wax Apple (Syzygium samarangense [Blume] Merrill & L. M. Perry var. Jambu Madu as Affected by Growth Regulator Application

    Directory of Open Access Journals (Sweden)

    Mohammad Moneruzzaman Khandaker

    2012-01-01

    Full Text Available This study represents the first paper of the effects of growth regulators on the physiochemical and phytochemical properties of the wax apple fruit, a widely cultivated fruit tree in southeast Asia. Net photosynthesis, sucrose phosphate synthase (SPS activity, peel color, fruit firmness, juice content, pH value, total soluble solids (TSSs, and the sugar acid ratio were all significantly increased in growth regulators (PGRs treated fruits. The application of gibberellin (GA3, naphthalene acetic acid (NAA, and 2,4-dichlorophenoxy acetic acid (2,4-D significantly reduced titratable acidity and increased total sugar and carbohydrate content compared to the control. The 50 mg/L GA3, 10 mg/L NAA, and 5 mg/L 2,4-D treatments produced the greatest increases in phenol and flavonoid content; vitamin C content was also higher for these treatments. PGR treatment significantly affected chlorophyll, anthocyanin, and carotene content and produced higher phenylalanine ammonia lyase (PAL and antioxidant activity levels. There was a positive correlation between peel color and TSS and antioxidant activity and both phenol and flavonoid content and PAL activity and anthocyanin formation. A taste panel assessment was also performed, and the highest scores were given to fruits that had been treated with GA3 or auxin. The study showed that application of 50 mg/L GA3, 10 mg/L NAA, and 5 mg/L 2,4-D once a week from bud development to fruit maturation increased the physiochemical and phytochemical properties of wax apple fruits.

  20. Characterization of the peptidylglycine α-amidating monooxygenase (PAM) from the venom ducts of neogastropods, Conus bullatus and Conus geographus.

    Science.gov (United States)

    Ul-Hasan, Sabah; Burgess, Daniel M; Gajewiak, Joanna; Li, Qing; Hu, Hao; Yandell, Mark; Olivera, Baldomero M; Bandyopadhyay, Pradip K

    2013-11-01

    Cone snails, genus Conus, are predatory marine snails that use venom to capture their prey. This venom contains a diverse array of peptide toxins, known as conotoxins, which undergo a diverse set of posttranslational modifications. Amidating enzymes modify peptides and proteins containing a C-terminal glycine residue, resulting in loss of the glycine residue and amidation of the preceding residue. A significant fraction of peptides present in the venom of cone snails contain C-terminal amidated residues, which are important for optimizing biological activity. This study describes the characterization of the amidating enzyme, peptidylglycine α-amidating monooxygenase (PAM), present in the venom duct of cone snails, Conus bullatus and Conus geographus. PAM is known to carry out two functions, peptidyl α-hydroxylating monooxygenase (PHM) and peptidylamido-glycolate lyase (PAL). In some animals, such as Drosophila melanogaster, these two functions are present in separate polypeptides, working as individual enzymes. In other animals, such as mammals and in Aplysia californica, PAM activity resides in a single, bifunctional polypeptide. Using specific oligonucleotide primers and reverse transcription-polymerase chain reaction we have identified and cloned from the venom duct cDNA library, a cDNA with 49% homology to PAM from A. californica. We have determined that both the PHM and PAL activities are encoded in one mRNA polynucleotide in both C. bullatus and C. geographus. We have directly demonstrated enzymatic activity catalyzing the conversion of dansyl-YVG-COOH to dansyl-YV-NH2 in cloned cDNA expressed in Drosophila S2 cells. Copyright © 2013 Elsevier Ltd. All rights reserved.

  1. Changes in energy metabolism accompanying pitting in blueberries stored at low temperature.

    Science.gov (United States)

    Zhou, Qian; Zhang, Chunlei; Cheng, Shunchang; Wei, Baodong; Liu, Xiuying; Ji, Shujuan

    2014-12-01

    Low-temperature storage and transport of blueberries is widely practiced in commercial blueberry production. In this research, the storage life of blueberries was extended at low temperature, but fruit stored for 30 d at 0°C pitted after 2d at room-temperature. Fruit cellular structure and physiological parameters accompanying pitting in blueberries were changed. The objective of this research was to characterise properties of energy metabolism accompanying pitting in blueberries during storage, including adenosine phosphates and mitochondrial enzymes involved in stress responses. Physiological and metabolic disorders, changes in cell ultrastructure, energy content and ATPase enzyme activity were observed in pitting blueberries. Energy shortages and increased activity of phenylalanine ammonia lyase (PAL) and lipoxygenase (LOX) were observed in fruit kept at shelf life. The results suggested that sufficient available energy status and a stable enzymatic system in blueberries collectively contribute to improve chilling tolerance, thereby alleviating pitting and maintaining quality of blueberry fruit in long-term cold storage. Copyright © 2014 Elsevier Ltd. All rights reserved.

  2. Review: Biflavonoid compounds of Selaginella Pal. Beauv. and its benefit

    Directory of Open Access Journals (Sweden)

    AHMAD DWI SETYAWAN

    2008-10-01

    Full Text Available In the present day, medicinal plants increasingly play important role in human health, while about 60-75% of world population depending on plants and their extracts for medication. Selaginella Pal. Beauv. (Selaginellaceae Reichb. is a potent medicinal plant source. This plant contains biflavonoid compounds, a dimeric form of flavonoids. Flavonoids are secondary metabolite that is most used in medical purposes. They are often occurred in daily consumed vegetables and fruits. Chemical constituents of flavonoids are very diverse, because they are generated in many biosynthetic methods. There are many biflavonoid compounds of Selaginella, i.e. amentoflavone, 2',8''-biapigenin, ginkgetin, heveaflavone, hinokiflavone, isocryptomerine, kayaflavone, podocarpusflavone A, robustaflavone, sumaflavone, and taiwaniaflavone. For the plants, these biflavonoid are used to response environmental condition such as defense against pests, diseases, herbivory, and competitions; while for the people, these compounds are used as antioxidant, anti-inflammatory, anti cancer, antimicrobial (antivirus, antibacterial, anti fungal, antiprotozoan, neuroprotective, vasorelaxant, anti UV-irradiation, antispasmodic, anti allergic, antihaemorrhagic, antinociceptive, etc. The antioxidant purpose of biflavonoid is the most important activities of this secondary metabolite. It is related to the anti cancer and anti-inflammatory functions. The antioxidant of biflavonoid is more powerful than β-carotene, vitamin C and E. In the future, Selaginella research exhaustively needs to be conducted on morphological and molecular characteristics. It deeply needs to explore on biflavonoid and other natural products constituents, and it also needs to search on their bioactivities.

  3. Effects of Salicylic Acid on the Growth and Physiological Characteristics in Cyclocarya Paliurus Seedlings

    International Nuclear Information System (INIS)

    Xie, Y.; Guo, N.; Zhang, Y.; Liu, N.; Wang, T.; Yang, W.; Fang, S.

    2016-01-01

    Field studies were conducted to examine the effects of salicylic acid (SA) on the growth and physiological characteristics of Cyclocarya paliurus seedlings by spraying the foliage with 0.0 (control), 0.2, 1.0, and 2.0 mM salicylic acid (SA). Proper concentrations of SA improved the relative growth yield of seedling stems and the soluble protein and sugar content of the leaves. It also increased the content of secondary metabolites including triterpenoids, flavonoids, quercetin and kaempferol, mineral elements K, Ca, Mg, Mn, Fe and Cu. Moreover, it stimulated the activities of superoxide dismutase (SOD), peroxidase (POX) and phenylalanine ammonia lyase (PAL) in the leaves. The effects of SA on these indices were dose dependent. The relative growth of seedling stem diameter and quercetin content increased gradually with an increase in concentration of SA from 0.0-2.0 mM. A concentration of 0.2 mM was optimal to promote content of soluble protein, sugars, Ca, Mg, Mn, Fe, and Cu, and SOD activity and significantly increased by 38.6 percentage, 22.1 percentage, 17.7 percentage, 8.2 percentage, 20.3 percentage, 23.2 percentage, 15.6 percentage, and 52.4 percentage, respectively, as compared with the control (CK). However, the maximal increase in activities of PAL, POX, and content of triterpenoids, kaempferol, and flavonoids was attained at 1.0 mM treatment, which significantly increased by 76.5 percentage, 78.4 percentage, 76.4 percentage, 96.3 percentage, and 107.4 percentage, respectively, compared with CK. Correlation analysis revealed positive relationships between activities of PAL, POX and content of triterpenoids, quercetin, kaempferol, and flavonoids within a certain concentration range of SA. These results suggested that an appropriate concentration (0.2-1.0 mM) of SA was not only effective in the improvement of physiological function of C. paliurus, but also increased seedling resistance; additionally, it helped to stimulate the synthesis of medicinal

  4. Joint Association of Dietary Pattern and Physical Activity Level with Cardiovascular Disease Risk Factors among Chinese Men: A Cross-Sectional Study.

    Directory of Open Access Journals (Sweden)

    Dong Wang

    Full Text Available The purpose of this cross-sectional study was to investigate the joint associations of physical activity level (PAL and dietary patterns in relation to cardiovascular disease (CVD risk factors among Chinese men. The study population consisted of 13 511 Chinese males aged 18-59 years from the 2002 China National Nutrition and Health Survey. Based on dietary data collected by a food frequency questionnaire, four dietary patterns were identified and labeled as "Green Water" (high consumption of rice, vegetables, seafood, pork, and poultry, "Yellow Earth" (high consumption of wheat flour products and starchy tubers, "New Affluent" (high consumption of animal sourced foods and soybean products, and "Western Adopter" (high consumption of animal sourced foods, cakes, and soft drinks. From the information collected by a 1-year physical activity questionnaire, PAL was calculated and classified into 4 categories: sedentary, low active, active, and very active. As compared with their counterparts from the New Affluent pattern, participants who followed the Green Water pattern had a lower likelihood of abdominal obesity (AO; 50.2%, hypertension (HT; 37.9%, hyperglycemia (HG; 41.5%, elevated triglyceride (ETG; 14.5%, low HDL (LHDL; 39.8%, and metabolic syndrome (MS; 51.9%. When compared to sedentary participants, the odds ratio of participants with very active PAL was 0.62 for AO, 0.85 for HT, 0.71 for HG, 0.76 for ETG, 0.74 for LHDL, and 0.58 for MS. Individuals who followed both very active PAL and the Green Water pattern had a lower likelihood of CVD risk factors (AO: 65.8%, HT: 39.1%, HG: 57.4%, ETG: 35.4%, LHDL: 56.1%, and MS: 75.0%, compared to their counterparts who followed both sedentary PAL and the New Affluent pattern. In addition, adherence to both healthy dietary pattern and very active PAL presented a remarkable potential for CVD risk factor prevention.

  5. Priming of seeds with methyl jasmonate induced resistance to hemi-biotroph Fusarium oxysporum f.sp. lycopersici in tomato via 12-oxo-phytodienoic acid, salicylic acid, and flavonol accumulation.

    Science.gov (United States)

    Król, P; Igielski, R; Pollmann, S; Kępczyńska, E

    2015-05-01

    Methyl jasmonate (MeJA) was tested by seed treatment for its ability to protect tomato seedlings against fusarium wilt caused by the soil-borne fungal pathogen Fusarium oxysporum f.sp. lycopersici. Isolated from Solanum lycopersicon L. seeds, cv. Beta fungus was identified as F. oxysporum f.sp. lycopersici Race 3 fungus by using phytopathological and molecular methods. MeJA applied at 0.01, 0.1 and 1 mM reduced spore germination and mycelial growth in vitro. Soaking of tomato seeds in MeJA solution at 0.1 mM for 1 h significantly enhanced the resistance level against the tested fungus in tomato seedlings 4 weeks after inoculation. The extracts from leaves of 15-day-old seedlings obtained from previously MeJA soaked seeds had the ability to inhibit in vitro spore germination of tested fungus. In these seedlings a significant increase in the levels phenolic compounds such as salicylic acid (SA), kaempferol and quercetin was observed. Up-regulation of phenylalanine ammonia-lyase (PAL5) and benzoic acid/salicylic acid carboxyl methyltransferase (BSMT) genes and down-regulation of the isochorysmate synthase (ICS) gene in response to exogenous MeJA application indicate that the phenylalanine ammonia-lyase (PAL), not the isochorismate (IC) pathway, is the primary route for SA production in tomato. Moreover, the increased accumulation of the flavonols quercetin and kaempferol appears closely related to the increase of PAL5, chalcone synthase (CHS) and flavonol synthase/flavanone 3-hydroxylase-like (FLS) genes. Elevated levels of salicylic acid in seedlings raised from MeJA-soaked seeds were simultaneously accompanied by a decrease of jasmonic acid, the precursor of MeJA, and an increase of 12-oxo-phytodienoic acid (OPDA), the precursor of jasmonic acid. The present results indicate that the priming of tomato seeds with 0.1mM MeJA before sowing enables the seedlings grown from these seeds to reduce the attack of the soil-borne fungal pathogen F. oxysporum f.sp. lycopersici

  6. Grazing-Activated Production of Dimethyl Sulfide (DMS) by two clones of Emiliania huxleyi

    Science.gov (United States)

    Wolfe, Gordon V.; Steinke, Michael

    1996-01-01

    Emiliania huxleyi clones CCMP 370 and CCMP 373 produced similar amounts of dimethylsulfoniopropionate (DMSP) during axenic exponential growth, averaging 109 mM internal DMSP. Both clones had detectable DMSP lyase activity, as measured by production of dimethyl sulfide (DMS) during in vitro assays of crude cell preparations, but activities and conditions differed considerably between clones. Clone 373 had high activity; clone 370 had low activity and required chloride. For both strains, enzyme activity per cell was constant during exponential growth, but little DMS was produced by healthy cells. Rather, DMS production was activated when cells were subjected to physical or chemical stresses that caused cell lysis. We propose that DMSP lyase and DMSP are segregated within these cells and re-action only under conditions that result in cell stress or damage. Such activation occurs during microzooplankton grazing. When these clones were grazed by the dinoflagellate Oxyrrhis marina, DMS was produced; ungrazed cells, as well as those exposed to grazer exudates and associated bacteria, generated no DMS. Grazing of clone 373 produced much more DMS than grazing of clone 370, consistent with their relative in vitro DMSP lyase activities. DMS was only generated when cells were actually being grazed, indicating that ingested cells were responsible for the DMS formation. We suggest that even low levels of grazing can greatly accelerate DMS production.

  7. Generation of 2-Furfurylthiol by Carbon-Sulfur Lyase from the Baijiu Yeast Saccharomyces cerevisiae G20.

    Science.gov (United States)

    Zha, Musu; Sun, Baoguo; Yin, Sheng; Mehmood, Arshad; Cheng, Lei; Wang, Chengtao

    2018-03-07

    2-Furfurylthiol is the representative aroma compound of Chinese sesame-flavored baijiu. Previous studies demonstrated that baijiu yeasts could generate 2-furfurylthiol using furfural and l-cysteine as precursors and that the Saccharomyces cerevisiae genes STR3 and CYS3 are closely related to 2-furfurylthiol biosynthesis. To confirm the mechanism of the STR3- and CYS3-gene products on 2-furfurylthiol biosynthesis, their encoded proteins were purified, and we confirmed their activities as carbon-sulfur lyases. Str3p and Cys3p were able to cleave the cysteine-furfural conjugate to release 2-furfurylthiol. Moreover, the characterization of the enzymatic properties of the purified proteins shows good thermal stabilities and wide pH tolerances, which enable their strong potential for various applications. These data provide direct evidence that yeast Str3p and Cys3p release 2-furfurylthiol in vitro, which can be applied to improve baijiu flavor.

  8. C12 derivatives of the hydroperoxide lyase pathway are produced by product recycling through lipoxygenase-2 in Nicotiana attenuata leaves

    NARCIS (Netherlands)

    Kallenbach, M.; Gilardoni, P.A.; Allmann, S.; Baldwin, I.T.; Bonaventure, G.

    2011-01-01

    In response to diverse stresses, the hydroperoxide lyase (HPL) pathway produces C(6) aldehydes and 12-oxo-(9Z )-dodecenoic acid ((9Z )-traumatin). Since the original characterization of (10E )-traumatin and traumatic acid, little has been added to our knowledge of the metabolism and fluxes

  9. [Association between cardiometabolic risk factors, physical activity and sedentariness in Chilean university students].

    Science.gov (United States)

    Morales Illanes, Gladys Ruth; Balboa-Castillo, Teresa; Muñoz, Sergio; Belmar, Carlos; Soto, Álvaro; Schifferli, Ingrid; Guillen-Grima, Francisco

    2017-11-16

    There is a huge evidence in adults, that a high level of physical activity and a low level of sedentariness are associated independently with a reduction of cardiometabolic risk factors (CMRF). This association has been poor evaluated in young people and the results are not conclusive. To estimate the association between CMRF, physical activity and sedentariness in Chilean university students. Cross-sectional study. Sample of 326 students who enrolled at University of La Frontera, Temuco, Chile, in 2014. They were selected randomly and stratified according to faculty, career and gender. We measured anthropometry, blood pressure, lipidic profile, glycemia, insulin resistance, tobacco and alcohol consumption. These variables were associated with physical activity level (PAL) and sedentariness using the IPAQ questionnaire. The students with a low PAL had a higher probability of abdominal obesity (OR: 4.68; IC 95% 1.86-11.73), metabolic syndrome (OR: 3.80; IC 95% 1.23-11.73) and triglycerides elevated (OR: 2.61 IC 95%; 1.18-5.75), compared with students with moderate and vigorous PAL (p < 0.05). There were no association between CMRF and sedentariness, even after fit by physical activity. We found an association between CMRF and PAL which was independent of sedentatiness. We believe it is very important to implement effective physical activity programs at universities. It is necesary to encourage students in order to increase the physical activity level to a moderate and vigorous range with the purpose of decreasing their cardiovascular risk. We consider it is recommendable to keep studying the association between sedentariness and CMRF.

  10. Unstable argininosuccinate lyase in variant forms of the urea cycle disorder argininosuccinic aciduria.

    Science.gov (United States)

    Hu, Liyan; Pandey, Amit V; Balmer, Cécile; Eggimann, Sandra; Rüfenacht, Véronique; Nuoffer, Jean-Marc; Häberle, Johannes

    2015-09-01

    Loss of function of the urea cycle enzyme argininosuccinate lyase (ASL) is caused by mutations in the ASL gene leading to ASL deficiency (ASLD). ASLD has a broad clinical spectrum ranging from life-threatening severe neonatal to asymptomatic forms. Different levels of residual ASL activity probably contribute to the phenotypic variability but reliable expression systems allowing clinically useful conclusions are not yet available. In order to define the molecular characteristics underlying the phenotypic variability, we investigated all ASL mutations that were hitherto identified in patients with late onset or mild clinical and biochemical courses by ASL expression in human embryonic kidney 293 T cells. We found residual activities >3% of ASL wild type (WT) in nine of 11 ASL mutations. Six ASL mutations (p.Arg95Cys, p.Ile100Thr, p.Val178Met, p.Glu189Gly, p.Val335Leu, and p.Arg379Cys) with residual activities ≥16% of ASL WT showed no significant or less than twofold reduced Km values, but displayed thermal instability. Computational structural analysis supported the biochemical findings by revealing multiple effects including protein instability, disruption of ionic interactions and hydrogen bonds between residues in the monomeric form of the protein, and disruption of contacts between adjacent monomeric units in the ASL tetramer. These findings suggest that the clinical and biochemical course in variant forms of ASLD is associated with relevant residual levels of ASL activity as well as instability of mutant ASL proteins. Since about 30% of known ASLD genotypes are affected by mutations studied here, ASLD should be considered as a candidate for chaperone treatment to improve mutant protein stability.

  11. Levels of potential bioactive compounds including carotenoids, vitamin C and phenolic compounds, and expression of their cognate biosynthetic genes vary significantly in different varieties of potato (Solanum tuberosum L.) grown under uniform cultural conditions.

    Science.gov (United States)

    Valcarcel, Jesus; Reilly, Kim; Gaffney, Michael; O'Brien, Nora M

    2016-02-01

    In addition to their high carbohydrate content, potatoes are also an important dietary source of vitamin C and bioactive secondary metabolites, including phenolic compounds and carotenoids, which have been suggested to play a role in human health. The expression of genes encoding key enzymes involved in the synthesis of these compounds was assessed by reverse transcription-quantitative polymerase chain reaction and compared to the accumulation of the corresponding product in seven potato varieties showing contrasting levels of metabolite accumulation. Strong positive correlations were found between phenolic content in the flesh of tubers and transcript levels of phenylalanine ammonia lyase (PAL) and chalcone synthase (CHS) genes. The expression of PAL and CHS was also related to that of AN1, a transcription factor involved in the synthesis of anthocyanins, suggesting that these genes are regulated in a coordinated manner. No clear relationship was found between transcript levels of phytoene synthase (PSY) or L-galactono-1,4-lactone dehydrogenase (GLDH) genes and total carotenoid or vitamin C accumulation, respectively. Data indicate that levels of total phenolic and flavonoid compounds in potato are controlled primarily by PAL and CHS gene expression. Transcript levels of PSY and GLDH did not control accumulation of carotenoids or vitamin C. © 2015 Society of Chemical Industry.

  12. Participation of the phosphoinositide metabolism in the hypersensitive response of Citrus limon against Alternaria alternata

    Directory of Open Access Journals (Sweden)

    XIMENA ORTEGA

    2001-01-01

    Full Text Available Lemon seedlings inoculated with Alternaria alternata develop a hypersensitive response (HR that includes the induction of Phenylalanine ammonia-lyase (PAL, E. C. 4.3.1.5 and the synthesis of scoparone. The signal transduction pathway involved in the development of this response is unknown. We used several inhibitors of the Phosphoinositide (PI animal system to study a possible role of Inositol-1,4,5-triphosphate (IP3 in the transduction of the fungal conidia signal in Citrus limon. The HR was only partially inhibited by EGTA, suggesting that not only external but internal calcium as well are necessary for a complete development of the HR. In this plant system, Alternaria alternata induced an early accumulation of the second messenger IP3. When lemon seedlings were watered long term with LiCl, an inhibitor of the phosphoinositide cycle, the IP3 production was reduced, and the LiCl-watered plants could neither induce PAL nor synthesize scoparone in response to fungal conidia. Furthermore, neomycin, a Phospholipase C (PLC, E. C. 3.1.4.3 inhibitor, also inhibited PAL induction and scoparone synthesis in response to A. alternata. These results suggest that IP3 could be involved in the signal transduction pathway for the development of the HR of Citrus limon against A. alternata

  13. Biochemical and physiological responses of oil palm to bud rot caused by Phytophthora palmivora.

    Science.gov (United States)

    Moreno-Chacón, Andrés Leonardo; Camperos-Reyes, Jhonatan Eduardo; Ávila Diazgranados, Rodrigo Andrés; Romero, Hernán Mauricio

    2013-09-01

    In recent years, global consumption of palm oil has increased significantly, reaching almost 43 million tons in 2010. The sustainability of oil palm (Elaeis guineensis) cultivation has been compromised because of the bud rot disease whose initial symptoms are caused by Phytophthora palmivora. There was a significant incidence of the disease, from an initial stage 1 of the disease to the highest stage 5, that affected photosynthetic parameters, content of pigments, sugars, polyamines, enzymatic antioxidant activities, phenylalanine ammonia-lyase (PAL, EC 4.3.1.5) and β-(1,3) glucanase (β-Gluc, EC 3.2.1.39). In healthy palms photosynthesis was 13.29 μmol CO2 m(-2) s(-1) in average, while in stage 5 the average photosynthesis was around 3.66 μmol CO2 m(-2) s(-1). Additionally, total chlorophyll was reduced by half at the last stage of the disease. On the contrary, the contents of putrescine, spermine and spermidine increased three, nine and twelve times with respect to stage 5, respectively. Antioxidant enzyme activities, as well as the phenylalanine ammonia-lyase and β-(1,3) glucanase showed an increase as the severity of the disease increased, with the latter increasing from 0.71 EAU in healthy palms to 2.60 EAU in plants at stage 5 of the disease. The peroxidase (POD, EC 1.11.1.7) enzymatic activity and the content of spermidine were the most sensitive indicators of disease. Copyright © 2013 Elsevier Masson SAS. All rights reserved.

  14. THE PAL 5 STAR STREAM GAPS

    International Nuclear Information System (INIS)

    Carlberg, R. G.; Hetherington, Nathan; Grillmair, C. J.

    2012-01-01

    Pal 5 is a low-mass, low-velocity-dispersion, globular cluster with spectacular tidal tails. We use the Sloan Digital Sky Survey Data Release 8 data to extend the density measurements of the trailing star stream to 23 deg distance from the cluster, at which point the stream runs off the edge of the available sky coverage. The size and the number of gaps in the stream are measured using a filter which approximates the structure of the gaps found in stream simulations. We find 5 gaps that are at least 99% confidence detections with about a dozen gaps at 90% confidence. The statistical significance of a gap is estimated using bootstrap resampling of the control regions on either side of the stream. The density minimum closest to the cluster is likely the result of the epicyclic orbits of the tidal outflow and has been discounted. To create the number of 99% confidence gaps per unit length at the mean age of the stream requires a halo population of nearly a thousand dark matter sub-halos with peak circular velocities above 1 km s –1 within 30 kpc of the galactic center. These numbers are a factor of about three below cold stream simulation at this sub-halo mass or velocity but, given the uncertainties in both measurement and more realistic warm stream modeling, are in substantial agreement with the LCDM prediction.

  15. A second pectin lyase gene (pel2) from Aspergillus oryzae KBN616: its sequence analysis and overexpression, and characterization of the gene products.

    Science.gov (United States)

    Kitamoto, N; Yoshino-Yasuda, S; Ohmiya, K; Tsukagoshi, N

    2001-01-01

    A second pectin lyase gene, designated pel2, was isolated from a shoyu koji mold Aspergillus oryzae KBN616 and characterized. The structural gene comprised 1306 bp with three introns. The ORF encoded 375 amino acids with a signal peptide of 19 amino acids. The deduced amino acid sequence showed high similarity to those of A. oryzae Pel1, Aspergillus niger pectin lyases and Glomerella cingulata Pn1A. The pel2 gene was overexpressed under the control of the promoter of the A. oryzae TEF1 gene for purification and enzymatic characterization of its gene product. The gene product exhibited two molecular masses of 48 and 44 kDa due to different degrees of glycosylation. Both proteins had the same pH optimum of 6.0 and temperature optimum of 50 degrees C.

  16. Electrochemistry of cytochrome P450 17α-hydroxylase/17,20-lyase (P450c17).

    Science.gov (United States)

    Martin, Lisandra L; Kubeil, Clemens; Simonov, Alexandr N; Kuznetsov, Vladimir L; Corbin, C Jo; Auchus, Richard J; Conley, Alan J; Bond, Alan M; Rodgers, Raymond J

    2017-02-05

    Within the superfamily of cytochrome P450 enzymes (P450s), there is a small class which is functionally employed for steroid biosynthesis. The enzymes in this class appear to have a small active site to accommodate the steroid substrates specifically and snuggly, prior to the redox transformation or hydroxylation to form a product. Cytochrome P450c17 is one of these and is also a multi-functional P450, with two activities, the first 17α-hydroxylation of pregnenolone is followed by a subsequent 17,20-lyase transformation to dehydroepiandrosterone (DHEA) as the dominant pathways to cortisol precursors or androgens in humans, respectively. How P450c17 regulates these two redox reactions is of special interest. There is a paucity of direct electrochemical studies on steroidogenic P450s, and in this mini-review we provide an overview of these studies with P450c17. Historical consideration as to the difficulties in obtaining reliable electrochemistry due to issues of handling proteins on an electrode, together with advances in the electrochemical techniques are addressed. Recent work using Fourier transformed alternating current voltammetry is highlighted as this technique can provide both catalytic information simultaneously with the underlying redox transfer with the P450 haem. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

  17. ÉTUDE DE CAS — Népal : La découverte de la diversité | IDRC ...

    International Development Research Centre (IDRC) Digital Library (Canada)

    5 janv. 2011 ... Agriculteurs et chercheurs travaillent à l'amélioration du riz et du maïs Les variétés locales de céréales cultivées dans les hautes terres du Népal pourraient faire beaucoup pour accroître les disponibilités alimentaires, mais elles ont été longtemps ignorées par le système officiel de la recherche.

  18. Physical activity, quality of life and medication in aging: differences between age and gender

    Directory of Open Access Journals (Sweden)

    Lilian Teresa Bucken Gobbi

    2009-04-01

    Full Text Available Studies have shown an inverse relationship between physical activity level (PAL, quality of life (QoL and use of medications in the elderly. The objective of this study was to analyze possible relationships and differences between PAL, QoL and use of medications in the elderly. A total of 192 subjects (≥ 60 years were selected by stratified random sampling according to census sector. The following assessment instruments were used: a Modified Baecke Questionnaire for older adults, b Medical Outcomes Study – 36-Item Short Form Health Survey, and c Sociodemographic and Health Factors Questionnaire. Descriptive statistics and parametric and nonparametric tests were used (p < 0.05. a With respect to chronological age, significant differences between groups were only observed for PAL, with G1 (60-69 years being more active than the other groups. b With respect to gender irrespective of age, analysis showed a difference in QoL and in the number of medications, with men reporting better perceived QoL and using fewer medications. c With respect to gender but considering chronological age, differences in PAL, QoL and medication use were observed between genders for specific age groups. In conclusion, in the elderly a PAL is low, declines even more during advanced age and is higher in men than in women during the first decade of old age, and b men report better perceived QoL and use fewer medications than women.

  19. Mini-review: recent developments in hydroxynitrile lyases for industrial biotechnology.

    Science.gov (United States)

    Lanfranchi, Elisa; Steiner, Kerstin; Glieder, Anton; Hajnal, Ivan; Sheldon, Roger A; van Pelt, Sander; Winkler, Margit

    2013-12-01

    Hydroxynitrile lyases (HNLs) catalyze the cleavage as well as the formation of cyanohydrins. The latter reaction is valuable for the stereoselective C-C bond formation by condensation of HCN with carbonyl compounds. The resulting cyanohydrins serve as versatile building blocks for a broad range of chemical and enzymatic follow-up reactions. A significant number of (R)- and (S)-selective HNLs are known today and the number is still increasing. HNLs not only exhibit varying substrate scope but also differ in sequence and structure. Tailor-made enzymes for large-scale manufacturing of cyanohydrins with improved yield and enantiomeric excess are very interesting targets, which is reflected in a solid number of patents. This review will complement and extend our recent review with a strong focus on applications of HNLs for the synthesis of highly functionalized, chiral compounds with newest literature, recent and current patent literature.

  20. Physical activity energy expenditure in Dutch adolescents: contribution of active transport to school, physical education, and leisure time activities.

    Science.gov (United States)

    Slingerland, Menno; Borghouts, Lars B; Hesselink, Matthijs K C

    2012-05-01

    Detailed knowledge about physical activity energy expenditure (PAEE) can guide the development of school interventions aimed at reducing overweight in adolescents. However, relevant components of PAEE have never been objectively quantified in this population. This study investigated the contribution of active transport to and from school, physical education (PE), and leisure time activities to total PAEE during a regular school week in adolescents. Seventy-three adolescents (mean age: 15.7 years) wore an individually calibrated combined heart rate-acceleration monitor and kept an activity diary during a regular school week. Branched equation modeling was used to calculate PAEE of the specific activity categories, and their relative contribution to total PAEE was determined. Active transport and PE contributed 30.0% and 17.4%, respectively, to school-related PAEE. Active transport to and from school contributed 15% to total PAEE. Youth with a high physical activity level (PAL) spent 4 hours less in sedentary behavior than subjects with a medium or low PAL (F = 77.415 (2.70), p activities (F = 10.583 (2.70), p Active transport and PE contribute significantly to PAEE during school hours in adolescents. To achieve an increase in total PAEE in the least active group of adolescents, promising strategies might be to reduce inactive behavior, increase participation in leisure time sports, and possibly to replace inactive for active jobs. © 2012, American School Health Association.

  1. Perceived need to increase physical activity levels among adults at high risk of type 2 diabetes. A cross-sectional analysis within a community-based diabetes prevention project FIN-D2D

    Directory of Open Access Journals (Sweden)

    Vähäsarja Kati

    2012-07-01

    Full Text Available Abstract Background Increased physical activity is a cornerstone of type 2 diabetes prevention. The perception of a need to change is considered essential in behaviour change processes. However, the existing literature on individuals’ perceived need to change health behaviour is limited. In order to improve understanding of diabetes prevention through increased physical activity levels (PAL, we assessed factors associated with perceiving a need to increase PAL among adults at high risk of diabetes. Methods Opportunistic screening was used within a primary-care based lifestyle intervention covering 10 149 men and women at high risk of type 2 diabetes. Data were obtained at baseline visits. The explored determinants were demographic, anthropometric/clinical, behavioural and psychosocial characteristics, along with four categories of PAL awareness. Logistic regression was used in the analysis. Results 74% of men (n = 2 577 and 76% of women (n = 4 551 perceived a need to increase their PAL. The participants most likely to perceive this need were inactive, had a larger waist circumference, rated their PAL as insufficient, and were at the contemplation stage of change. Smoking, elevated blood pressure, dyslipidaemia, and a family history of diabetes were not associated with this perception. The likelihood was also greater among women with less perceived fitness and less education. Demographic factors other than education did not determine participants’ perceived need to increase PAL. PAL overestimators were less likely to perceive the need to increase their PAL than realistic inactive participants. Conclusions Subjective rather than objective health factors appear to determine the perception of a need to increase PAL among adults at high risk of diabetes. Client perceptions need to be evaluated in health counselling in order to facilitate a change in PAL. Practical descriptions of the associations between metabolic risk factors, PAL, and

  2. Extraction and Separation of Active Ingredients in Schisandra chinensis (Turcz. Baill and the Study of their Antifungal Effects.

    Directory of Open Access Journals (Sweden)

    Haijing Yi

    Full Text Available Schisandra chinensis extracts (SEs have traditionally been used as an oriental medicine for the treatment of various human diseases, however, their further application in the biocontrol of plant disease remains poorly understood. This study was conducted to develop eco-friendly botanical pesticides from extracts of S. chinensis and assess whether they could play a key role in plant disease defense. Concentrated active fractions (SE-I, SE-II, and SE-III were obtained from S. chinensis via specific extraction and separation. Then, lignan-like substances, such as Schisanhenol B, were detected via High-Performance Liquid Chromatography-ElectroSpray Ionization-Mass Spectrometry (HPLC-ESI-MS analyses of the active fractions. Moreover, the results from biological tests on colony growth inhibition and spore germination indicated that SE-I, SE-II, and SE-III could inhibit hyphal growth and spore generation of three important plant pathogenic fungi (Monilinia fructicola, Fusarium oxysporum, and Botryosphaeria dothidea. The study of the mechanisms of resistant fungi revealed that the oxidation resistance system, including reactive oxygen species (ROS, malondialdehyde (MDA, catalase (CAT, and superoxide dismutase (SOD, was activated. The expression of genes related to defense, such as pathogenesis-related protein (PR4, α-farnesene synthase (AFS, polyphenol oxidase (PPO, and phenylalanine ammonia lyase (PAL were shown to be up-regulated after treatment with SEs, which suggested an increase in apple immunity and that fruits were induced to effectively defend against the infection of pathogenic fungi (B. dothidea. This study revealed that SEs and their lignans represent promising resources for the development of safe, effective, and multi-targeted agents against pathogenic fungi.

  3. Extraction and Separation of Active Ingredients in Schisandra chinensis (Turcz.) Baill and the Study of their Antifungal Effects

    Science.gov (United States)

    Liu, Jun; Zhang, Jie; Guo, Wei; Xiao, Weilie; Yao, Yuncong

    2016-01-01

    Schisandra chinensis extracts (SEs) have traditionally been used as an oriental medicine for the treatment of various human diseases, however, their further application in the biocontrol of plant disease remains poorly understood. This study was conducted to develop eco-friendly botanical pesticides from extracts of S. chinensis and assess whether they could play a key role in plant disease defense. Concentrated active fractions (SE-I, SE-II, and SE-III) were obtained from S. chinensis via specific extraction and separation. Then, lignan-like substances, such as Schisanhenol B, were detected via High-Performance Liquid Chromatography-ElectroSpray Ionization-Mass Spectrometry (HPLC-ESI-MS) analyses of the active fractions. Moreover, the results from biological tests on colony growth inhibition and spore germination indicated that SE-I, SE-II, and SE-III could inhibit hyphal growth and spore generation of three important plant pathogenic fungi (Monilinia fructicola, Fusarium oxysporum, and Botryosphaeria dothidea). The study of the mechanisms of resistant fungi revealed that the oxidation resistance system, including reactive oxygen species (ROS), malondialdehyde (MDA), catalase (CAT), and superoxide dismutase (SOD), was activated. The expression of genes related to defense, such as pathogenesis-related protein (PR4), α-farnesene synthase (AFS), polyphenol oxidase (PPO), and phenylalanine ammonia lyase (PAL) were shown to be up-regulated after treatment with SEs, which suggested an increase in apple immunity and that fruits were induced to effectively defend against the infection of pathogenic fungi (B. dothidea). This study revealed that SEs and their lignans represent promising resources for the development of safe, effective, and multi-targeted agents against pathogenic fungi. PMID:27152614

  4. Potential role of pectate lyase and Ca(2+) in the increase in strawberry fruit firmness induced by short-term treatment with high-pressure CO2.

    Science.gov (United States)

    Wang, Mao Hua; Kim, Jin Gook; Ahn, Sun Eun; Lee, Ah Youn; Bae, Tae Min; Kim, Deu Re; Hwang, Yong Soo

    2014-04-01

    Postharvest treatment with high-pressure CO2 helps to control decay and increase firmness in strawberries. Increases in firmness occurred through modification of calcium binding to cell wall. However, the mechanism(s) involved in Ca(2+) migration to pectic polymers and other physiological events associated with the maintenance of increased firmness are not clearly understood. The focus of this study was to find potential mechanism(s) that are associated with calcium movement, increases in firmness, or maintenance of firmness in strawberry fruit after high-pressure CO2 treatment. An increase in firmness was induced by high-pressure CO2 treatment, but not by high-pressure N2 treatment. This indicates that CO2 stimulates a change in firmness. The increase in firmness induced by high-pressure CO2 seems to involve calcium efflux. Using membrane Ca(2+) -dependent ATPase inhibitors sodium vanadate (250 μM) and erythrosin B (100 μM) delayed both the increase in firmness and calcium binding to wall polymers. Exogenous application of CaCl2 (10 mM) enhanced the firmness increase of fruit slices only when they were exposed to high-pressure CO2 . The activity of pectate lyase was downregulated by CO2 treatment, but β-galactosidase activity was not affected. The increase in strawberry firmness induced by high-pressure CO2 treatment primarily involves the efflux of calcium ions and their binding to wall polymers. These physiological changes are not induced by an anaerobic environment. The downregulation of wall-modifying enzymes, such as pectate lyase, appeared to contribute to the maintenance of firmness that was induced by high-pressure CO2 treatment. © 2014 Institute of Food Technologists®

  5. Kinetic resolution and stereoselective synthesis of 3-substituted aspartic acids by using engineered methylaspartate ammonia lyases.

    Science.gov (United States)

    Raj, Hans; Szymanski, Wiktor; de Villiers, Jandré; Puthan Veetil, Vinod; Quax, Wim J; Shimamoto, Keiko; Janssen, Dick B; Feringa, Ben L; Poelarends, Gerrit J

    2013-08-19

    Enzymatic amino acid synthesis: Kinetic resolution and asymmetric synthesis of various valuable 3-substituted aspartic acids, which were obtained in fair to good yields with diastereomeric ratio values of up to >98:2 and enantiomeric excess values of up to >99 %, by using engineered methylaspartate ammonia lyases are described. These biocatalytic methodologies for the selective preparation of aspartic acid derivatives appear to be attractive alternatives for existing chemical methods. Copyright © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  6. Solidago canadensis L essential oil vapor effectively inhibits Botrytis cinerea growth and preserves postharvest quality of strawberry as a food model system

    Directory of Open Access Journals (Sweden)

    Shumin Liu

    2016-08-01

    Full Text Available This study investigated the anti-fungal properties of Solidago canadensis L essential oil (SCLEO against Botrytis cinerea in vitro, and its ability to control gray mold and maintain quality in strawberry fruits. SCLEO exhibited dose-dependent antifungal activity against B. cinerea and profoundly altered mycelial morphology, cellular ultrastructure, and membrane permeability as evaluated by scanning electron microscopy, transmission electron microscopy, and fluorescence microscopy. SCLEO vapor at 0.1 mL/L maintained higher sensory acceptance and reduced decay of fresh strawberry fruit, and also reduced gray mold in artificially inoculated fruit. SCLEO treatment did not however, stimulate phenylalanin ammonia-lyase (PAL, polyphenol oxidase (POD, or chitinase (CHI, enzymes related to disease resistance. This suggests that SCLEO reduces gray mold by direct inhibition of pathogen growth. SCLEO vapor may provide a new and effective strategy for controlling postharvest disease and maintaining quality in strawberries.

  7. Effect of foliar application of salicylic acid, hydrogen peroxide and a xyloglucan oligosaccharide on capsiate content and gene expression associatedwith capsinoids synthesis in Capsicum annuum L.

    Science.gov (United States)

    Zunun-Perez, A Y; Guevara-Figueroa, T; Jimenez-Garcia, S N; Feregrino-Perez, A A; Gautier, F; Guevara-Gonzalez, R G

    2017-06-01

    Capsinoids are non-pungent analogues of capsaicinoids in pepper (Capsicum spp). The absence of pungency, in addition to their biological activities similar to that of capsaicinoids such as anti-inflammatory, antimicrobial, and antioxidant properties, makes capsinoids an excellent option for increasing use in human and animal nutrition, as well as health and pharmaceutical industries. There are only few sources of pepper producing capsinoids, and one of them (accession 509-45-1), Capsicum annuum L., is a potential source for increasing capsinoids content using strategies as controlled elicitation during plant production in the greenhouse. In this research we evaluated the effect of weekly and one-day-before-harvest foliar applications of hydrogen peroxide, salicylic acid and a xyloglucan oligosaccharide on the concentration of capsiate in fruits of this pepper accession, as well as the gene expression of phenylalanine ammonia-lyase (pal), putative aminotransferase (pamt), capsaicin synthase (at3) and β-keto acyl synthase (kas). Results showed that the two tested concentrations of H2O2 significantly increased capsiate content and gene expression associated with capsaicinoids (pamt, at3 and kas) and the phenylpropanoids (pal) pathways. Plant yield was not affected using this induction strategy. Our results indicated that the pre-harvest and weekly application of hydrogen peroxide and xyloglucan oligosaccharide improved production of capsiate in C. annuum L.

  8. Mutant form C115H of Clostridium sporogenes methionine γ-lyase efficiently cleaves S-Alk(en)yl-l-cysteine sulfoxides to antibacterial thiosulfinates.

    Science.gov (United States)

    Kulikova, Vitalia V; Anufrieva, Natalya V; Revtovich, Svetlana V; Chernov, Alexander S; Telegin, Georgii B; Morozova, Elena A; Demidkina, Tatyana V

    2016-10-01

    Pyridoxal 5'-phosphate-dependent methionine γ-lyase (MGL) catalyzes the β-elimination reaction of S-alk(en)yl-l-cysteine sulfoxides to thiosulfinates, which possess antimicrobial activity. Partial inactivation of the enzyme in the course of the reaction occurs due to oxidation of active site cysteine 115 conserved in bacterial MGLs. In this work, the C115H mutant form of Clostridium sporogenes MGL was prepared and the steady-state kinetic parameters of the enzyme were determined. The substitution results in an increase in the catalytic efficiency of the mutant form towards S-substituted l-cysteine sulfoxides compared to the wild type enzyme. We used a sulfoxide/enzyme system to generate antibacterial activity in situ. Two-component systems composed of the mutant enzyme and three S-substituted l-cysteine sulfoxides were demonstrated to be effective against Gram-positive and Gram-negative bacteria and three clinical isolates from mice. © 2016 IUBMB Life, 68(10):830-835, 2016. © 2016 International Union of Biochemistry and Molecular Biology.

  9. Catalytic-site mapping of pyruvate formate lyase. Hypophosphite reaction on the acetyl-enzyme intermediate affords carbon-phosphorus bond synthesis (1-hydroxyethylphosphonate).

    Science.gov (United States)

    Plaga, W; Frank, R; Knappe, J

    1988-12-15

    Pyruvate formate-lyase of Escherichia coli cells, a homodimeric protein of 2 x 85 kDa, is distinguished by the property of containing a stable organic free radical (g = 2.0037) in its resting state. The enzyme (E-SH) achieves pyruvate conversion to acetyl-CoA via two distinct half-reactions (E-SH + pyruvate in equilibrium E-S-acetyl + formate; E-S-acetyl + CoA in equilibrium E-SH + acetyl-CoA), the first of which has been proposed to involve reversible homolytic carbon-carbon bond cleavage [J. Knappe et al. (1984) Proc. Natl Acad. Sci. USA 81, 1332-1335]. Present studies identified Cys-419 as the covalent-catalytic cysteinyl residue via CNBr fragmentation of E-S-[14C]acetyl and radio-sequencing of the isolated peptide CB-Ac (amino acid residues 406-423). Reaction of the formate analogue hypophosphite with E-S-acetyl was investigated and found to produce 1-hydroxyethylphosphonate with a thioester linkage to the adjacent Cys-418. The structure was determined from the chymotryptic peptide CH-P (amino acid residues 415-425), using 31P-NMR spectroscopy (delta = 44 ppm) and by chemical characterisation through degradation into 1-hydroxyethylphosphonate with phosphodiesterase or bromine. This novel P-C-bond synthesis involves the enzyme-based free radical and is proposed to resemble the physiological C-C-bond synthesis (pyruvate production) from formate and E-S-acetyl. These findings are interpreted as proof of a radical mechanism for the action of pyruvate formate-lyase. The central Cys-418/Cys-419 pair of the active site shows a distinctive thiolate property even in the inactive (nonradical) form of the enzyme, as determined using an iodoacetate probe.

  10. Crosslinked enzyme aggregates of hydroxynitrile lyase partially purified from Prunus dulcis seeds and its application for the synthesis of enantiopure cyanohydrins.

    Science.gov (United States)

    Yildirim, Deniz; Tükel, S Seyhan; Alagöz, Dilek

    2014-01-01

    Hydroxynitrile lyases are powerful catalysts in the synthesis of enantiopure cyanohydrins which are key synthons in the preparations of a variety of important chemicals. The response surface methodology including three-factor and three-level Box-Behnken design was applied to optimize immobilization of hydroxynitrile lyase purified partially from Prunus dulcis seeds as crosslinked enzyme aggregates (PdHNL-CLEAs). The quadratic model was developed for predicting the response and its adequacy was validated with the analysis of variance test. The optimized immobilization parameters were initial glutaraldehyde concentration, ammonium sulfate saturation concentration, and crosslinking time, and the response was relative activity of PdHNL-CLEA. The optimal conditions were determined as initial glutaraldehyde concentration of 25% w/v, ammonium sulfate saturation concentration of 43% w/v, and crosslinking time of 18 h. The preparations of PdHNL-CLEA were examined for the synthesis of (R)-mandelonitrile, (R)-2-chloromandelonitrile, (R)-3,4-dihydroxymandelonitrile, (R)-2-hydroxy-4-phenyl butyronitrile, (R)-4-bromomandelonitrile, (R)-4-fluoromandelonitrile, and (R)-4-nitromandelonitrile from their corresponding aldehydes and hydrocyanic acid. After 96-h reaction time, the yield-enantiomeric excess values (%) were 100-99, 100-21, 100-99, 83-91, 100-99, 100-72, and 100-14%, respectively, for (R)-mandelonitrile, (R)-2-chloromandelonitrile, (R)-3,4-dihydroxymandelonitrile, (R)-2-hydroxy-4-phenyl butyronitrile, (R)-4-bromomandelonitrile, (R)-4-fluoromandelonitrile, and (R)-4-nitromandelonitrile. The results show that PdHNL-CLEA offers a promising potential for the preparation of enantiopure (R)-mandelonitrile, (R)-3,4-dihydroxymandelonitrile, (R)-2-hydroxy-4-phenyl butyronitrile, and (R)-4-bromomandelonitrile with a high yield and enantiopurity. © 2014 American Institute of Chemical Engineers.

  11. Purification and characterization of selenocysteine beta-lyase from Citrobacter freundii

    International Nuclear Information System (INIS)

    Chocat, P.; Esaki, N.; Tanizawa, K.; Nakamura, K.; Tanaka, H.; Soda, K.

    1985-01-01

    The purification and characterization of bacterial selenocysteine beta-lyase, an enzyme which specifically catalyzes the cleavage of L-selenocysteine to L-alanine and Se0, are presented. The enzyme, purified to near homogeneity from Citrobacter freundii, is monomeric with a molecular weight of ca. 64,000 and contains 1 mol of pyridoxal 5'-phosphate as a cofactor per mol of enzyme. L-Selenocysteine is the sole substrate. L-Cysteine is a competitive inhibitor of the enzyme. The enzyme also catalyzes the alpha, beta elimination of beta-chloro-L-alanine to form NH 3 , pyruvate, and Cl- and is irreversibly inactivated during the reaction. The physicochemical properties, e.g., amino acid composition and subunit structure, of the bacterial enzyme are fairly different from those of the pig liver enzyme. However, the catalytic properties of both enzymes, e.g., substrate specificity and inactivation by the substrate or a mechanism-based inactivator, beta-chloro-L-alanine, are very similar

  12. Comparison of silicon nanoparticles and silicate treatments in fenugreek.

    Science.gov (United States)

    Nazaralian, Sanam; Majd, Ahmad; Irian, Saeed; Najafi, Farzaneh; Ghahremaninejad, Farrokh; Landberg, Tommy; Greger, Maria

    2017-06-01

    Silicon (Si) fertilization improves crop cultivation and is commonly added in the form of soluble silicates. However, most natural plant-available Si originates from plant formed amorphous SiO 2 particles, phytoliths, similar to SiO 2 -nanoparticles (SiNP). In this work we, therefore, compared the effect by sodium silicate and that of SiNP on Si accumulation, activity of antioxidative stress enzymes catalase, peroxidase, superoxide dismutase, lignification of xylem cell walls and activity of phenylalanine ammonia-lyase (PAL) as well as expression of genes for the putative silicon transporter (PST), defensive (Tfgd 1) and phosphoenolpyruvate carboxykinase (PEPCK) and protein in fenugreek (Trigonella foenum-graecum L.) grown in hydroponics. The results showed that Si was taken up from both silicate and SiNP treatments and increasing sodium silicate addition increased the translocation of Si to the shoot, while this was not shown with increasing SiNP addition. The silicon transporter PST was upregulated at a greater level when sodium silicate was added compared with SiNP addition. There were no differences in effects between sodium silicate and SiNP treatments on the other parameters measured. Both treatments increased the uptake and accumulation of Si, xylem cell wall lignification, cell wall thickness, PAL activity and protein concentration in seedlings, while there was no effect on antioxidative enzyme activity. Tfgd 1 expression was strongly downregulated in leaves at Si addition. The similarity in effects by silicate and SiNP would be due to that SiNP releases silicate, which may be taken up, shown by a decrease in SiNP particle size with time in the medium. Copyright © 2017 Elsevier Masson SAS. All rights reserved.

  13. Effect of ultraviolet-C light on postharvest decay of strawberry

    International Nuclear Information System (INIS)

    Nigro, F.; Ippolito, A.; Salerno, M.; Lattanzio, V.

    2000-01-01

    The effect of ultraviolet-C light (u.v.-C) at low doses on postharvest decay of strawberries caused by Botrytis cinerea and other pathogens was investigated. Phenylalanine ammonia-lyase (PAL) activity and ethylene production, as influenced by ultraviolet-C irradiation, were also determined. Strawberries (cv. ‘Pajaro’) from plants that had been treated with chemicals against grey mould were irradiated with u.v.-C doses ranging from 0.25 to 4.00 kJ m-2 and inoculated with B. cinerea at different times (0, 12, 24 and 48 hours) after irradiation. To assess the effect of u.v.-C light on the naturally occurring postharvest decay, organically grown strawberries were also used. After treatment the strawberries were stored at 20±1°C or at 3°C. u.v.-C doses at 0.50 and 1.00 kJ m-2 significantly reduced botrytis storage rot arising from both artificial inoculations and natural infections in comparison with the unirradiated control. The doses shown to reduce botrytis rot produced an increase in PAL activity 12 h after irradiation; this result indicates the activation of metabolic a pathway related to the biosynthesis of phenolic compounds, which are usually characterized by antifungal activity. In addition, u.v.-C irradiation caused an increase in ethylene production proportional to the doses applied, reaching the highest value 6 h after treatment. The overall results from these investigations indicate that treatment with low u.v.-C doses produces a reduction in postharvest decay of strawberries related to induced resistance mechanisms. Moreover, a germicidal effect of reducing external contaminating pathogens cannot be excluded

  14. Reforming pathology teaching in medical college by peer-assisted learning and student-oriented interest building activities: A pilot study.

    Science.gov (United States)

    Grover, Sumit; Sood, Neena; Chaudhary, Anurag

    2017-01-01

    Peer-assisted learning (PAL) is a teaching-learning method in which students act as peer teachers and help other students to learn while also themselves learning by teaching. PAL through modified interest building activities (MIBAs) is seldom tried in teaching pathology in medical colleges. This study aimed to evaluate the usefulness of peer teaching using MIBA, obtain feedback from students, and compare different activities with each other and with traditional teaching-learning methods. An interventional pilot study was conducted in 2 months on the 2nd MBBS undergraduates learning pathology at a medical college in North India. Students acted as peer teachers and performed different MIBAs including role plays, demonstration of pathogenesis through props, student-led seminars such as PowerPoint teaching, blackboard teaching, multiple choice question seminars, case-based learning (CBL) exercises, and quizzes before teaching sessions. Feedback was obtained through structured questionnaires on a 5-point Likert scale. Paired t-test was used to compare traditional teaching with MIBAs, and Friedman test was used to compare among different MIBAs. Students found ease of understanding and the interaction and involvement of students as the most important benefits of PAL. MIBAs increased voluntary participation, coordination, teamwork, shared responsibility, and group dynamics among students. Quiz sessions followed by PowerPoint seminars and prop demonstrations received highest mean scores from students on most of the parameters. Quizzes, blackboard teaching, prop activities, and CBL helped students understand topics better and generated interest. Learners advocated for making MIBAs and PAL compulsory for future students. PAL complemented by MIBAs may be adopted to make teaching-learning more interesting and effective through the active involvement and participation of students.

  15. Physical activity, quality of life and medication in aging: differences between age and gender

    Directory of Open Access Journals (Sweden)

    Émerson Sebastião

    2009-01-01

    Full Text Available http://dx.doi.org/10.5007/1980-0037.2009v11n2p210   Studies have shown an inverse relationship between physical activity level (PAL, quality of life (QoL and use of medications in the elderly. The objective of this study was to analyze possible relationships and differences between PAL, QoL and use of medications in the elderly. A total of 192 subjects (≥ 60 years were selected by stratified random sampling according to census sector. The following assessment instruments were used: a Modified Baecke Questionnaire for older adults, b Medical Outcomes Study – 36-Item Short Form Health Survey, and c Sociodemographic and Health Factors Questionnaire. Descriptive statistics and parametric and nonparametric tests were used (p < 0.05. a With respect to chronological age, significant differences between groups were only observed for PAL, with G1 (60-69 years being more active than the other groups. b With respect to gender irrespective of age, analysis showed a difference in QoL and in the number of medications, with men reporting better perceived QoL and using fewer medications. c With respect to gender but considering chronological age, differences in PAL, QoL and medication use were observed between genders for specific age groups. In conclusion, in the elderly a PAL is low, declines even more during advanced age and is higher in men than in women during the first decade of old age, and b men report better perceived QoL and use fewer medications than women.

  16. General Aviation Activity and Avionics Survey 1984

    Science.gov (United States)

    1985-10-01

    1-r- 0sn r-C) 1-n4 A rW IniC lol o oo i.VA Cd )’ a .3 -- - I, CdA xd j Cd ’Am. " ’ Ai I-W wU g t 8 weR at lz at w betl wg at i w~ a w~ at ix 8 OM I0at...S ’ TABLE D-1. SDR AIRCRAFT GROUP NAME - FAA MANUFACTURER/MODEL CODES (CONTINUED) SDR FAA SDR FAA SDR FAA PIPER 600 106001... PIPER PAlS 101828 PIPER PA31T 103128 PIPER 600 106010 PIPER PAl8 101832 PIPER PA32 103206 PIPER 600 106012 PIPER PAlS 101834 PIPER PA32 103207 PIPER 600

  17. Comparison of the Danish Physical Activity Questionnaire with a validated position and motion instrument

    DEFF Research Database (Denmark)

    Matthiessen, Jeppe; Biltoft-Jensen, Anja Pia; Rasmussen, Lone Banke

    2008-01-01

    Objective To compare the Danish Physical Activity Questionnaire (DPAQ) estimating physical activity energy expenditure (PAEE) and physical activity level (PAL) and the pattern of physical activity (including health-related physical activity) with measurements from a validated position and motion ...

  18. Sexual Dimorphism in the Selenocysteine Lyase Knockout Mouse.

    Science.gov (United States)

    Ogawa-Wong, Ashley N; Hashimoto, Ann C; Ha, Herena; Pitts, Matthew W; Seale, Lucia A; Berry, Marla J

    2018-01-31

    Selenium (Se) is an essential micronutrient known for its antioxidant properties and health benefits, attributed to its presence in selenoproteins as the amino acid, selenocysteine. Selenocysteine lyase (Scly) catalyzes hydrolysis of selenocysteine to selenide and alanine, facilitating re-utilization of Se for de novo selenoprotein synthesis. Previously, it was reported that male Scly -/- mice develop increased body weight and body fat composition, and altered lipid and carbohydrate metabolism, compared to wild type mice. Strikingly, females appeared to present with a less severe phenotype, suggesting the relationship between Scly and energy metabolism may be regulated in a sex-specific manner. Here, we report that while body weight and body fat gain occur in both male and female Scly -/- mice, strikingly, males are susceptible to developing glucose intolerance, whereas female Scly -/- mice are protected. Because Se is critical for male reproduction, we hypothesized that castration would attenuate the metabolic dysfunction observed in male Scly -/- mice by eliminating sequestration of Se in testes. We report that fasting serum insulin levels were significantly reduced in castrated males compared to controls, but islet area was unchanged between groups. Finally, both male and female Scly -/- mice exhibit reduced hypothalamic expression of selenoproteins S, M, and glutathione peroxidase 1.

  19. Sexual Dimorphism in the Selenocysteine Lyase Knockout Mouse

    Directory of Open Access Journals (Sweden)

    Ashley N. Ogawa-Wong

    2018-01-01

    Full Text Available Selenium (Se is an essential micronutrient known for its antioxidant properties and health benefits, attributed to its presence in selenoproteins as the amino acid, selenocysteine. Selenocysteine lyase (Scly catalyzes hydrolysis of selenocysteine to selenide and alanine, facilitating re-utilization of Se for de novo selenoprotein synthesis. Previously, it was reported that male Scly−/− mice develop increased body weight and body fat composition, and altered lipid and carbohydrate metabolism, compared to wild type mice. Strikingly, females appeared to present with a less severe phenotype, suggesting the relationship between Scly and energy metabolism may be regulated in a sex-specific manner. Here, we report that while body weight and body fat gain occur in both male and female Scly−/− mice, strikingly, males are susceptible to developing glucose intolerance, whereas female Scly−/− mice are protected. Because Se is critical for male reproduction, we hypothesized that castration would attenuate the metabolic dysfunction observed in male Scly−/− mice by eliminating sequestration of Se in testes. We report that fasting serum insulin levels were significantly reduced in castrated males compared to controls, but islet area was unchanged between groups. Finally, both male and female Scly−/− mice exhibit reduced hypothalamic expression of selenoproteins S, M, and glutathione peroxidase 1.

  20. Neuroprotective effects of (Val8)GLP-1-Glu-PAL in the MPTP Parkinson's disease mouse model

    OpenAIRE

    Zhang, YanFang; Chen, YiMei; Li, Lin; Holscher, Christian

    2015-01-01

    Glucagon-like peptide 1 (GLP-1) is a hormone and a growth factor. GLP-1 mimetics are currently on the market as treatments for type 2 diabetes. They also have shown neuroprotective properties in animal models of neurodegenerative disorders. In addition, the GLP-1 mimetic exendin-4 has shown protective effects in animal models of Parkinson's disease (PD), and a first clinical trial in PD patients showed promising results. (Val8)GLP-1-glu-PAL is a new GLP-1 analogue which has a longer biologica...

  1. Energy requirements and physical activity level of active elderly people in rural areas of Cuba. Highlights and achievements

    International Nuclear Information System (INIS)

    Hernandez-Trianna, M.

    2002-01-01

    Physical activity levels (PALs) values of urban elderly of developed countries obtained by the DLW-method are usually higher than the recommendations of the Expert Committee on Energy and Protein. This underestimation should be more evident for the rural elderly

  2. Application of Osthol Induces a Resistance Response Against Powdery Mildew in Pumpkin Leave

    Directory of Open Access Journals (Sweden)

    Yong Jian Fan

    2007-09-01

    Full Text Available Plants can defend themselves against fungal infection by natural means inducedby biotic and abiotic elicitors. Osthol is a natural compound extracted from dried fruits ofCnidii Monnieri Fructus. In this study, it has been shown to not only be a fungicide withacceptable curative properties (control efficacy of 68.72, but it also showed a significantprophylactic effect (with control efficacy of 77.36 against pumpkin powdery mildew at aconcentration of 100 μg·mL-1. In pumpkin leaves with/or without inoculation ofSphaerotheca fuliginea, osthol treatment induced the accumulation of chitinase andperoxidase and enhanced the transcription of chitinase gene in non-inoculated leaves. Thepotentiation of phenylalanine amonia-lyase activity in leaves by osthol application andfollowing inoculation was absent in that with inoculation or osthol treatment, indicatingthat induced PAL in osthol-pretreated plants was inoculation-mediated. In conclusion, thisnatural compound could induce resistance response in the plant against powdery mildew.

  3. Engineering of Escherichia coli for the synthesis of N-hydroxycinnamoyl tryptamine and serotonin.

    Science.gov (United States)

    Lee, Su Jin; Sim, Geun-Young; Lee, Youngshim; Kim, Bong-Gyu; Ahn, Joong-Hoon

    2017-11-01

    Plants synthesize various phenol amides. Among them, hydroxycinnamoyl (HC) tryptamines and serotonins exhibit antioxidant, anti-inflammatory, and anti-atherogenic activities. We synthesized HC-tryptamines and HC-serotonin from several HCs and either tryptamine or serotonin using Escherichia coli harboring the 4CL (4-coumaroyl CoA ligase) and CaHCTT [hydroxycinnamoyl-coenzyme A:serotonin N-(hydroxycinnamoyl)transferase] genes. E. coli was engineered to synthesize N-cinnamoyl tryptamine from glucose. TDC (tryptophan decarboxylase) and PAL (phenylalanine ammonia lyase) along with 4CL and CaHCTT were introduced into E. coli and the phenylalanine biosynthetic pathway of E. coli was engineered. Using this strategy, approximately 110.6 mg/L of N-cinnamoyl tryptamine was synthesized. By feeding 100 μM serotonin into the E. coli culture, which could induce the synthesis of cinnamic acid or p-coumaric acid, more than 99 μM of N-cinnamoyl serotonin and N-(p-coumaroyl) serotonin were synthesized.

  4. ATP-Citrate Lyase Controls a Glucose-to-Acetate Metabolic Switch

    Directory of Open Access Journals (Sweden)

    Steven Zhao

    2016-10-01

    Full Text Available Mechanisms of metabolic flexibility enable cells to survive under stressful conditions and can thwart therapeutic responses. Acetyl-coenzyme A (CoA plays central roles in energy production, lipid metabolism, and epigenomic modifications. Here, we show that, upon genetic deletion of Acly, the gene coding for ATP-citrate lyase (ACLY, cells remain viable and proliferate, although at an impaired rate. In the absence of ACLY, cells upregulate ACSS2 and utilize exogenous acetate to provide acetyl-CoA for de novo lipogenesis (DNL and histone acetylation. A physiological level of acetate is sufficient for cell viability and abundant acetyl-CoA production, although histone acetylation levels remain low in ACLY-deficient cells unless supplemented with high levels of acetate. ACLY-deficient adipocytes accumulate lipid in vivo, exhibit increased acetyl-CoA and malonyl-CoA production from acetate, and display some differences in fatty acid content and synthesis. Together, these data indicate that engagement of acetate metabolism is a crucial, although partial, mechanism of compensation for ACLY deficiency.

  5. ATP-Citrate Lyase Controls a Glucose-to-Acetate Metabolic Switch.

    Science.gov (United States)

    Zhao, Steven; Torres, AnnMarie; Henry, Ryan A; Trefely, Sophie; Wallace, Martina; Lee, Joyce V; Carrer, Alessandro; Sengupta, Arjun; Campbell, Sydney L; Kuo, Yin-Ming; Frey, Alexander J; Meurs, Noah; Viola, John M; Blair, Ian A; Weljie, Aalim M; Metallo, Christian M; Snyder, Nathaniel W; Andrews, Andrew J; Wellen, Kathryn E

    2016-10-18

    Mechanisms of metabolic flexibility enable cells to survive under stressful conditions and can thwart therapeutic responses. Acetyl-coenzyme A (CoA) plays central roles in energy production, lipid metabolism, and epigenomic modifications. Here, we show that, upon genetic deletion of Acly, the gene coding for ATP-citrate lyase (ACLY), cells remain viable and proliferate, although at an impaired rate. In the absence of ACLY, cells upregulate ACSS2 and utilize exogenous acetate to provide acetyl-CoA for de novo lipogenesis (DNL) and histone acetylation. A physiological level of acetate is sufficient for cell viability and abundant acetyl-CoA production, although histone acetylation levels remain low in ACLY-deficient cells unless supplemented with high levels of acetate. ACLY-deficient adipocytes accumulate lipid in vivo, exhibit increased acetyl-CoA and malonyl-CoA production from acetate, and display some differences in fatty acid content and synthesis. Together, these data indicate that engagement of acetate metabolism is a crucial, although partial, mechanism of compensation for ACLY deficiency. Copyright © 2016 The Authors. Published by Elsevier Inc. All rights reserved.

  6. Evaluation of Borage Extracts As Potential Biostimulant Using a Phenomic, Agronomic, Physiological, and Biochemical Approach.

    Science.gov (United States)

    Bulgari, Roberta; Morgutti, Silvia; Cocetta, Giacomo; Negrini, Noemi; Farris, Stefano; Calcante, Aldo; Spinardi, Anna; Ferrari, Enrico; Mignani, Ilaria; Oberti, Roberto; Ferrante, Antonio

    2017-01-01

    Biostimulants are substances able to improve water and nutrient use efficiency and counteract stress factors by enhancing primary and secondary metabolism. Premise of the work was to exploit raw extracts from leaves (LE) or flowers (FE) of Borago officinalis L., to enhance yield and quality of Lactuca sativa 'Longifolia,' and to set up a protocol to assess their effects. To this aim, an integrated study on agronomic, physiological and biochemical aspects, including also a phenomic approach, has been adopted. Extracts were diluted to 1 or 10 mL L -1 , sprayed onto lettuce plants at the middle of the growing cycle and 1 day before harvest. Control plants were treated with water. Non-destructive analyses were conducted to assess the effect of extracts on biomass with an innovative imaging technique, and on leaf photosynthetic efficiency (chlorophyll a fluorescence and leaf gas exchanges). At harvest, the levels of ethylene, photosynthetic pigments, nitrate, and primary (sucrose and total sugars) and secondary (total phenols and flavonoids) metabolites, including the activity and levels of phenylalanine ammonia lyase (PAL) were assessed. Moreover, a preliminary study of the effects during postharvest was performed. Borage extracts enhanced the primary metabolism by increasing leaf pigments and photosynthetic activity. Plant fresh weight increased upon treatments with 10 mL L -1 doses, as correctly estimated by multi-view angles images. Chlorophyll a fluorescence data showed that FEs were able to increase the number of active reaction centers per cross section; a similar trend was observed for the performance index. Ethylene was three-fold lower in FEs treatments. Nitrate and sugar levels did not change in response to the different treatments. Total flavonoids and phenols, as well as the total protein levels, the in vitro PAL specific activity, and the levels of PAL-like polypeptides were increased by all borage extracts, with particular regard to FEs. FEs also proved

  7. Evaluation of Borage Extracts As Potential Biostimulant Using a Phenomic, Agronomic, Physiological, and Biochemical Approach

    Directory of Open Access Journals (Sweden)

    Roberta Bulgari

    2017-06-01

    Full Text Available Biostimulants are substances able to improve water and nutrient use efficiency and counteract stress factors by enhancing primary and secondary metabolism. Premise of the work was to exploit raw extracts from leaves (LE or flowers (FE of Borago officinalis L., to enhance yield and quality of Lactuca sativa ‘Longifolia,’ and to set up a protocol to assess their effects. To this aim, an integrated study on agronomic, physiological and biochemical aspects, including also a phenomic approach, has been adopted. Extracts were diluted to 1 or 10 mL L–1, sprayed onto lettuce plants at the middle of the growing cycle and 1 day before harvest. Control plants were treated with water. Non-destructive analyses were conducted to assess the effect of extracts on biomass with an innovative imaging technique, and on leaf photosynthetic efficiency (chlorophyll a fluorescence and leaf gas exchanges. At harvest, the levels of ethylene, photosynthetic pigments, nitrate, and primary (sucrose and total sugars and secondary (total phenols and flavonoids metabolites, including the activity and levels of phenylalanine ammonia lyase (PAL were assessed. Moreover, a preliminary study of the effects during postharvest was performed. Borage extracts enhanced the primary metabolism by increasing leaf pigments and photosynthetic activity. Plant fresh weight increased upon treatments with 10 mL L–1 doses, as correctly estimated by multi-view angles images. Chlorophyll a fluorescence data showed that FEs were able to increase the number of active reaction centers per cross section; a similar trend was observed for the performance index. Ethylene was three-fold lower in FEs treatments. Nitrate and sugar levels did not change in response to the different treatments. Total flavonoids and phenols, as well as the total protein levels, the in vitro PAL specific activity, and the levels of PAL-like polypeptides were increased by all borage extracts, with particular regard to FEs

  8. Phytochemical study of phenolic compounds of labrador tea (Lédum palústre L.

    Directory of Open Access Journals (Sweden)

    Валентина Петровна Гапоненко

    2015-11-01

    Full Text Available Aim. This work is devoted a phytochemical study of biologically active substances in herb of Labrador Tea (Lédum palústre L., as well as investigation of the possibility of complex use of raw materials in order to create on its basis new herbal medicines.Methods. The object of the study served as the herb Labrador Tea. Separation of isolated substances was performed by adsorption and partition chromatography on various adsorbents. The structure of the isolated compounds was determined based on physicochemical methods: paper (PC and the thin layer (TLC chromatography, UV, IR and NMR spectroscopy in comparison with the original valid standards of flavonoids. Acid hydrolys was used for the determination of the flavonoid aglycone composition. The content of total flavonoids was determined by differential spectrophotometry at a wavelength of 412 nm from the reaction with aluminum chloride based on the hyperoside-standard (Ukrainian scientific Pharmacopoeial center for quality of medicines (Pharmacopoeial center, Kharkоv.Results. During this study we found more than 40 of phenolic compounds and identified 31compounds. The phytochemical analysis showed the presence of flavonols (11 compounds, catechins (5. Hydroxycinnamic acids represented by caffeic, ferulic, chlorogenic acids. Besides that, it were found phenolic glycoside arbutin, coumarins - coumarin, umbelliferon, scopoletin, esculetin and esculin, tannins – metyl gallate, pyrogallol.Conclusions. The following biologically active substances were defined for the first time: flavonoids – 5-methyl-kaempferol, avicularin, polistahozid, quercitrin; coumarins – esculetin, esculin; as well as hydroxycinnamic acids – ferulic, chlorogenic, neochlorogenic of Labrador Tea. The obtained data justify the prospectivefor creation of new and effective herbal medicines from Labrador Tea

  9. The crystal structures of the tri-functional Chloroflexus aurantiacus and bi-functional Rhodobacter sphaeroides malyl-CoA lyases and comparison with CitE-like superfamily enzymes and malate synthases.

    Science.gov (United States)

    Zarzycki, Jan; Kerfeld, Cheryl A

    2013-11-09

    Malyl-CoA lyase (MCL) is a promiscuous carbon-carbon bond lyase that catalyzes the reversible cleavage of structurally related Coenzyme A (CoA) thioesters. This enzyme plays a crucial, multifunctional role in the 3-hydroxypropionate bi-cycle for autotrophic CO2 fixation in Chloroflexus aurantiacus. A second, phylogenetically distinct MCL from Rhodobacter sphaeroides is involved in the ethylmalonyl-CoA pathway for acetate assimilation. Both MCLs belong to the large superfamily of CitE-like enzymes, which includes the name-giving β-subunit of citrate lyase (CitE), malyl-CoA thioesterases and other enzymes of unknown physiological function. The CitE-like enzyme superfamily also bears sequence and structural resemblance to the malate synthases. All of these different enzymes share highly conserved catalytic residues, although they catalyze distinctly different reactions: C-C bond formation and cleavage, thioester hydrolysis, or both (the malate synthases). Here we report the first crystal structures of MCLs from two different phylogenetic subgroups in apo- and substrate-bound forms. Both the C. aurantiacus and the R. sphaeroides MCL contain elaborations on the canonical β8/α8 TIM barrel fold and form hexameric assemblies. Upon ligand binding, changes in the C-terminal domains of the MCLs result in closing of the active site, with the C-terminal domain of one monomer forming a lid over and contributing side chains to the active site of the adjacent monomer. The distinctive features of the two MCL subgroups were compared to known structures of other CitE-like superfamily enzymes and to malate synthases, providing insight into the structural subtleties that underlie the functional versatility of these enzymes. Although the C. aurantiacus and the R. sphaeroides MCLs have divergent primary structures (~37% identical), their tertiary and quaternary structures are very similar. It can be assumed that the C-C bond formation catalyzed by the MCLs occurs as proposed for

  10. Effects of truncation of the peptide chain on the secondary structure and bioactivities of palmitoylated anoplin.

    Science.gov (United States)

    Salas, Remmer L; Garcia, Jan Kathryne D L; Miranda, Ana Carmela R; Rivera, Windell L; Nellas, Ricky B; Sabido, Portia Mahal G

    2018-06-01

    Anoplin (GLLKRIKTLL-NH 2 ) is of current interest due to its short sequence and specificity towards bacteria. Recent studies on anoplin have shown that truncation and acylation compromises its antimicrobial activity and specificity, respectively. In this study, truncated analogues (pal-ano-9 to pal-ano-5) of palmitoylated anoplin (pal-anoplin) were synthesized to determine the effects of C-truncation on its bioactivities. Moreover, secondary structure of each analogue using circular dichroism (CD) spectroscopy was determined to correlate with bioactivities. Interestingly, pal-anoplin, pal-ano-9 and pal-ano-6 were helical in water, unlike anoplin. In contrast, pal-ano-8, pal-ano-7 and pal-ano-5, with polar amino acid residues at the C-terminus, were random coil in water. Nevertheless, all the peptides folded into helical structures in 30% trifluoroethanol/water (TFE/H 2 O) except for the shortest analogue pal-ano-5. Hydrophobicity played a significant role in the enhancement of activity against bacteria E. coli and S. aureus as all lipopeptides including the random coil pal-ano-5 were more active than the parent anoplin. Meanwhile, the greatest improvement in activity against the fungus C. albicans was observed for pal-anoplin analogues (pal-ano-9 and pal-ano-6) that were helical in water. Although, hydrophobicity is a major factor in the secondary structure and antimicrobial activity, it appears that the nature of amino acids at the C-terminus also influence folding of lipopeptides in water and its antifungal activity. Moreover, the hemolytic activity of the analogues was found to correlate with hydrophobicity, except for the least hemolytic, pal-ano-5. Since most of the analogues are more potent and shorter than anoplin, they are promising drug candidates for further development. Copyright © 2018. Published by Elsevier Inc.

  11. NFAT regulation of cystathionine γ-lyase expression in endothelial cells is impaired in rats exposed to intermittent hypoxia.

    Science.gov (United States)

    Gonzalez Bosc, Laura V; Osmond, Jessica M; Giermakowska, Wieslawa K; Pace, Carolyn E; Riggs, Jennifer L; Jackson-Weaver, Olan; Kanagy, Nancy L

    2017-04-01

    Sleep apnea is a risk factor for cardiovascular disease, and intermittent hypoxia (IH, 20 episodes/h of 5% O 2 -5% CO 2 for 7 h/day) to mimic sleep apnea increases blood pressure and impairs hydrogen sulfide (H 2 S)-induced vasodilation in rats. The enzyme that produces H 2 S, cystathionine γ-lyase (CSE), is decreased in rat mesenteric artery endothelial cells (EC) following in vivo IH exposure. In silico analysis identified putative nuclear factor of activated T cell (NFAT) binding sites in the CSE promoter. Therefore, we hypothesized that IH exposure reduces Ca 2+ concentration ([Ca 2+ ]) activation of calcineurin/NFAT to lower CSE expression and impair vasodilation. In cultured rat aortic EC, inhibiting calcineurin with cyclosporine A reduced CSE mRNA, CSE protein, and luciferase activity driven by a full-length but not a truncated CSE promoter. In male rats exposed to sham or IH conditions for 2 wk, [Ca 2+ ] in EC in small mesenteric arteries from IH rats was lower than in EC from sham rat arteries (Δfura 2 ratio of fluorescence at 340 to 380 nm from Ca 2+ free: IH = 0.05 ± 0.02, sham = 0.17 ± 0.03, P intermittent hypoxia to mimic sleep apnea, nuclear factor of activated T cells c3 nuclear translocation and CSE expression are decreased, concomitant with decreased CSE-dependent vasodilation. Copyright © 2017 the American Physiological Society.

  12. Volume changes at macro- and nano-scale in epoxy resins studied by PALS and PVT experimental techniques

    Energy Technology Data Exchange (ETDEWEB)

    Somoza, A. [IFIMAT-UNCentro, Pinto 399, B7000GHG Tandil (Argentina) and CICPBA, Pinto 399, B7000GHG Tandil (Argentina)]. E-mail: asomoza@exa.unicen.edu.ar; Salgueiro, W. [IFIMAT-UNCentro, Pinto 399, B7000GHG Tandil (Argentina); Goyanes, S. [LPMPyMC, Depto. de Fisica, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Ciudad Universitaria, Pabellon I, 1428 Buenos Aires (Argentina); Ramos, J. [Materials and Technology Group, Departamento de Ingenieria Quimica y M. Ambiente, Escuela University Politecnica, Universidad Pais Vasco/Euskal Herriko Unibertsitatea, Pz. Europa 1, 20018 Donostia/San Sebastian (Spain); Mondragon, I. [Materials and Technology Group, Departamento de Ingenieria Quimica y M. Ambiente, Escuela University Politecnica, Universidad Pais Vasco/Euskal Herriko Unibertsitatea, Pz. Europa 1, 20018 Donostia/San Sebastian (Spain)

    2007-02-15

    A systematic study on changes in the volumes at macro- and nano-scale in epoxy systems cured with selected aminic hardeners at different pre-cure temperatures is presented. Free- and macroscopic specific-volumes were measured by PALS and pressure-volume-temperature techniques, respectively. An analysis of the relation existing between macro- and nano-scales of the thermosetting networks developed by the different chemical structures is shown. The result obtained indicates that the structure of the hardeners governs the packing of the molecular chains of the epoxy network.

  13. Aerobic exercise did not have compensatory effects on physical activity levels in type 2 diabetes patients.

    Science.gov (United States)

    de Moura, Bruno Pereira; Marins, João Carlos Bouzas; Franceschini, Sylvia Do Carmo Castro; Reis, Janice Sepúlveda; Amorim, Paulo Roberto Dos Santos

    2015-01-01

    Although exercise promotes beneficial effects in diabetic patients, some studies have questioned the degree of their importance in terms of the increase in total energy expenditure. In these studies, the decrease of physical activity levels (PAL) was referred as "compensatory effect of exercise". However, our aim was to investigate whether aerobic exercise has compensatory effects on PAL in type 2 diabetes patients. Eight volunteers (51.1 ± 8.2 years) were enrolled in a supervised exercise programme for 8 weeks (3 d · wk(-1), 50-60% of VO2 peak for 30-60 min). PAL was measured using tri-axial accelerometers in the 1st, 8th and 12th weeks. Biochemical tests, cardiorespiratory fitness, anthropometric assessment and body composition were measured in the 2nd and 11th weeks. Statistical analysis was performed using non-parametric tests (Friedman and Wilcoxon, P exercise programme generated a significant 14.8% increase in VO2 peak and a 15% reduction in fructosamine. The exercise programme had no compensatory effects on PAL in type 2 diabetes patients, but improved their cardiorespiratory fitness and glycaemic control.

  14. Real time expression of ACC oxidase and PR-protein genes mediated by Methylobacterium spp. in tomato plants challenged with Xanthomonas campestris pv. vesicatoria.

    Science.gov (United States)

    Yim, W J; Kim, K Y; Lee, Y W; Sundaram, S P; Lee, Y; Sa, T M

    2014-07-15

    Biotic stress like pathogenic infection increases ethylene biosynthesis in plants and ethylene inhibitors are known to alleviate the severity of plant disease incidence. This study aimed to reduce the bacterial spot disease incidence in tomato plants caused by Xanthomonas campestris pv. vesicatoria (XCV) by modulating stress ethylene with 1-aminocyclopropane-1-carboxylate (ACC) deaminase activity of Methylobacterium strains. Under greenhouse condition, Methylobacterium strains inoculated and pathogen challenged tomato plants had low ethylene emission compared to pathogen infected ones. ACC accumulation and ACC oxidase (ACO) activity with ACO related gene expression increased in XCV infected tomato plants over Methylobacterium strains inoculated plants. Among the Methylobacterium spp., CBMB12 resulted lowest ACO related gene expression (1.46 Normalized Fold Expression), whereas CBMB20 had high gene expression (3.42 Normalized Fold Expression) in pathogen challenged tomato. But a significant increase in ACO gene expression (7.09 Normalized Fold Expression) was observed in the bacterial pathogen infected plants. In contrast, Methylobacterium strains enhanced β-1,3-glucanase and phenylalanine ammonia-lyase (PAL) enzyme activities in pathogen challenged tomato plants. The respective increase in β-1,3-glucanase related gene expressions due to CBMB12, CBMB15, and CBMB20 strains were 66.3, 25.5 and 10.4% higher over pathogen infected plants. Similarly, PAL gene expression was high with 0.67 and 0.30 Normalized Fold Expression, in pathogen challenged tomato plants inoculated with CBMB12 and CBMB15 strains. The results suggest that ethylene is a crucial factor in bacterial spot disease incidence and that methylobacteria with ACC deaminase activity can reduce the disease severity with ultimate pathogenesis-related protein increase in tomato. Copyright © 2014 Elsevier GmbH. All rights reserved.

  15. Late Neolithic vegetation history at the pile-dwelling site of Palù di Livenza (northeastern Italy

    Science.gov (United States)

    Pini, Roberta

    2004-12-01

    The Late Neolithic pile-dwelling of Palù di Livenza yielded archaeological remains typical of the Square Mouth Pottery and Lagozza Cultures. A palynological investigation reveals important changes in the vegetation due to anthropogenic pressure. Between ca. 6590 and 5960 cal. yr BP, dense oak wood forests with deciduous Quercus, Fagus and Corylus extended around the mire, with no signs of human impact. The establishment of the pile-dwelling, dated to ca. 5960 cal. yr BP, led to a strong reduction of forests, reclamation of wetlands, and expansion of herbaceous communities, with cultivated species, infestant weeds, nitrophilous and ruderal herbs, pastures and meadows. According to AMS dates and previous archaeological chronologies, the pile-dwelling persisted for about 700 years (from ca. 5960 to 5260 cal. yr BP). The history of the pile-dwelling after ca. 5260 cal. yr BP cannot be reconstructed because of recent contamination of the top part of the section. Rarefaction analysis was applied to estimate changes of palynological richness through time: the highest E(Tn) (between 56 and 69 taxa) are contemporaneous with the local development of the pile-dwelling. The comparison of pollen data with archaeobotanical evidence indicates that Fragaria vesca, Malus sylvestris, Papaver somniferum and Physalis alkekengi were gathered at some distance from the site and that Linum usitatissimum is strongly under-represented in pollen samples. Crop cultivation can be estimated for a radius of several hundred metres around the mire. Palù di Livenza is significant in the context of Neolithic archaeobotany of northern Italy and neighbouring countries. Copyright

  16. Design, Synthesis, and Biological Evaluation of Isothiosemicarbazones with Antimycobacterial Activity

    Czech Academy of Sciences Publication Activity Database

    Novotná, E.; Waisser, K.; Kuneš, J.; Palát, K.; Skálová, L.; Szotáková, B.; Buchta, V.; Stolaříková, J.; Ulmann, V.; Pávová, Marcela; Weber, Jan; Komrsková, J.; Hašková, P.; Vokřál, I.; Wsól, V.

    2017-01-01

    Roč. 350, č. 8 (2017), č. článku e1700020. ISSN 0365-6233 Institutional support: RVO:61388963 Keywords : biological activity * cytotoxicity * isocitrate lyase * isothiosemicarbazone * tuberculosis Subject RIV: EE - Microbiology, Virology OBOR OECD: Microbiology Impact factor: 1.994, year: 2016

  17. Measurement of the target current by inductive probe during laser interaction on terawatt laser system PALS

    Czech Academy of Sciences Publication Activity Database

    Cikhardt, Jakub; Krása, Josef; De Marco, Massimo; Pfeifer, Miroslav; Velyhan, Andriy; Krouský, Eduard; Cikhardtová, B.; Klír, Daniel; Řezáč, Karel; Ullschmied, Jiří; Skála, Jiří; Kubeš, P.; Kravárik, J.

    2014-01-01

    Roč. 85, č. 10 (2014), s. 103507-103507 ISSN 0034-6748 R&D Projects: GA MŠk LM2010014; GA MŠk(CZ) LG13029; GA ČR GAP205/12/0454; GA MŠk EE2.3.20.0279 Grant - others:LaserZdroj (OP VK 3)(XE) CZ.1.07/2.3.00/20.0279 Institutional support: RVO:61389021 ; RVO:68378271 Keywords : laser PALS * laser-target interaction * target current * inductive probe Subject RIV: BL - Plasma and Gas Discharge Physics; BL - Plasma and Gas Discharge Physics (FZU-D) Impact factor: 1.614, year: 2014 http://dx.doi.org/10.1063/1.4898016

  18. Levels of cystathionine gamma lyase production by Geotrichum candidum in synthetic media and correlation with the presence of sulphur flavours in cheese.

    Science.gov (United States)

    Gente, Stéphanie; La Carbona, Stéphanie; Guéguen, Micheline

    2007-03-10

    Geotrichum candidum is a cheese-ripening agent with the potential to produce sulphur flavour compounds in soft cheeses. We aimed to develop an alternative test for predicting the aromatic (sulphur flavours) potential of G. candidum strains in soft cheese. Twelve strains of G. candidum with different levels of demethiolase activity (determined by a chemical method) in YEL-met (yeast extract, lactate methionine) medium were studied. We investigated cgl (cystathionine gamma lyase) gene expression after culture in three media - YEL-met, casamino acid and curd media - and then carried out sensory analysis on a Camembert cheese matrix. We found no correlation between demethiolase activity in vitro and cgl gene expression. Sensory analysis (detection of sulphur flavours) identified different aromatic profiles linked to cgl expression, but not to demethiolase activity. The RT-PCR technique described here is potentially useful for predicting the tendency of a given strain of G. candidum to develop sulphur flavours in cheese matrix. This is the first demonstration that an in vitro molecular approach could be used as a predictive test for evaluating the potential of G. candidum strains to generate sulphur compounds in situ (Camembert cheese matrix).

  19. Peer assisted learning in the clinical setting: an activity systems analysis

    OpenAIRE

    Bennett, Deirdre; O?Flynn, Siun; Kelly, Martina

    2014-01-01

    Peer assisted learning (PAL) is a common feature of medical education. Understanding of PAL has been based on processes and outcomes in controlled settings, such as clinical skills labs. PAL in the clinical setting, a complex learning environment, requires fresh evaluation. Socio-cultural theory is proposed as a means to understand educational interventions in ways that are practical and meaningful. We describe the evaluation of a PAL intervention, introduced to support students? transition i...

  20. Passive aquatic listener (PAL): An adoptive underwater acoustic recording system for the marine environment

    International Nuclear Information System (INIS)

    Anagnostou, Marios N.; Nystuen, Jeffrey A.; Anagnostou, Emmanouil N.; Papadopoulos, Anastasios; Lykousis, Vassilios

    2011-01-01

    The ambient sound field in the ocean is a combination of natural and manmade sounds. Consequently, the interpretation of the ambient sound field can be used to quantify these processes. In the frequency range from 1 to 50 kHz, the general character of ocean ambient sound is a slowly changing background that is closely associated with local wind speed, interspersed with shorter time scale events such as rain storms, ships and animal calls. At lower frequencies the underwater ambient sound budget includes geologically generated sound activities including underwater volcanic eruptions, seismic and seepage faults that generate bubbles, etc. that can also potentially be classified and quantified. Acoustic data are collected on hydrophones. Hydrophones are simple, robust sensors that can be deployed on most ocean instrumentation systems including surface or sub-surface moorings, bottom mounted systems, drifters, ARGO floats or autonomous underwater platforms. A dedicated oceanic underwater recorder called a passive acoustic listener (PAL) has been developed. A principal issue is to accurately distinguish different sound sources so that they can be quantified as part of a sound budget, and then quantified if appropriate. Based on ongoing data collected from the Poseidon II network the retrieval potential of multi-parameters from underwater sound, including meteorological (i.e., precipitation and winds) and in general geophysical, anthropogenetic (i.e., ships, submarines, etc.) and biological (whales, etc.) sources is presented.

  1. Passive aquatic listener (PAL): An adoptive underwater acoustic recording system for the marine environment

    Energy Technology Data Exchange (ETDEWEB)

    Anagnostou, Marios N., E-mail: managnostou@ath.hcmr.g [Hellenic Center for Marine Research, Department of Inland Waters, Anavissos (Greece); Nystuen, Jeffrey A. [Hellenic Center for Marine Research, Department of Inland Waters, Anavissos (Greece); Applied Physics Laboratory, University of Washington, Seattle, WA (United States); Anagnostou, Emmanouil N. [Hellenic Center for Marine Research, Department of Inland Waters, Anavissos (Greece); Department of Civil and Environmental Engineering, University of Connecticut, CT (United States); Papadopoulos, Anastasios [Hellenic Center for Marine Research, Department of Inland Waters, Anavissos (Greece); Lykousis, Vassilios [Hellenic Center for Marine Research, Department of Oceanography, Anavissos (Greece)

    2011-01-21

    The ambient sound field in the ocean is a combination of natural and manmade sounds. Consequently, the interpretation of the ambient sound field can be used to quantify these processes. In the frequency range from 1 to 50 kHz, the general character of ocean ambient sound is a slowly changing background that is closely associated with local wind speed, interspersed with shorter time scale events such as rain storms, ships and animal calls. At lower frequencies the underwater ambient sound budget includes geologically generated sound activities including underwater volcanic eruptions, seismic and seepage faults that generate bubbles, etc. that can also potentially be classified and quantified. Acoustic data are collected on hydrophones. Hydrophones are simple, robust sensors that can be deployed on most ocean instrumentation systems including surface or sub-surface moorings, bottom mounted systems, drifters, ARGO floats or autonomous underwater platforms. A dedicated oceanic underwater recorder called a passive acoustic listener (PAL) has been developed. A principal issue is to accurately distinguish different sound sources so that they can be quantified as part of a sound budget, and then quantified if appropriate. Based on ongoing data collected from the Poseidon II network the retrieval potential of multi-parameters from underwater sound, including meteorological (i.e., precipitation and winds) and in general geophysical, anthropogenetic (i.e., ships, submarines, etc.) and biological (whales, etc.) sources is presented.

  2. Descriptive and predictive assessment of enzyme activity and enzyme related processes in biorefinery using IR spectroscopy and chemometrics

    DEFF Research Database (Denmark)

    Baum, Andreas

    the understanding of the structural properties of the extracted pectin. Secondly, enzyme kinetics of biomass converting enzymes was examined in terms of measuring enzyme activity by spectral evolution profiling utilizing FTIR. Chemometric multiway methods were used to analyze the tensor datasets enabling the second......-order calibration advantage (reference Theory of Analytical chemistry). As PAPER 3 illustrates the method is universally applicable without the need of any external standards and was exemplified by performing quantitative enzyme activity determinations for glucose oxidase, pectin lyase and a cellolytic enzyme blend...... (Celluclast 1.5L). In PAPER 4, the concept is extended to quantify enzyme activity of two simultaneously acting enzymes, namely pectin lyase and pectin methyl esterase. By doing so the multiway methods PARAFAC, TUCKER3 and NPLS were compared and evaluated towards accuracy and precision....

  3. Physical activity level and sedentary behavior among students

    Directory of Open Access Journals (Sweden)

    Roberto Jerônimo dos Santos Silva

    2009-06-01

    Full Text Available The objective of this study was to determine the physical activity level (PAL and sedentary behavior of students from the city of Aracaju (SE. A total of 1028 students of both genders participated in the study, with a mean age of 15.38 (2.44 years for girls and 15.24 (2.40 years for boys. Among the sample, 24.7% were children and 75.3% were adolescents, with a mean age of 12.07 (0.88 and 16.39 (1.72 years, respectively. An instrument already used in Brazilian studies was applied to identify the average time (hours watching TV per day (hTV and PAL – PAQ-C. Descriptive statistics, t-test for independent samples, Fisher’s exact test and comparison test between two proportions were used for data analysis, with the level of significance set at 5% (p≤0.05. Boys presented a significantly higher physical activity score 2.25 (0.60 than girls. The prevalence of sedentarism was 72.5, 89.3 and 85.2% in the groups of children and adolescents and in the group as a whole, respectively, for girls, and 55.4, 74.8 and 69.8% for boys. No differences in hTV were observed between genders or between sedentary and physically active students (p > 0.05. We conclude that a there is a high prevalence of “sedentary” and “very sedentary” children and adolescents; b boys present a higher PAL than girls; c adolescents are less active than children, and d the number of hTV is high in the group studied.

  4. Adenylosuccinate lyase (ADSL) and infantile autism: Absence of previously reported point mutation

    Energy Technology Data Exchange (ETDEWEB)

    Fon, E.A.; Sarrazin, J.; Rouleau, G.A. [Montreal General Hospital (Canada)] [and others

    1995-12-18

    Autism is a heterogeneous neuropsychiatric syndrome of unknown etiology. There is evidence that a deficiency in the enzyme adenylosuccinate lyase (ADSL), essential for de novo purine biosynthesis, could be involved in the pathogenesis of certain cases. A point mutation in the ADSL gene, resulting in a predicted serine-to-proline substitution and conferring structural instability to the mutant enzyme, has been reported previously in 3 affected siblings. In order to determine the prevalence of the mutation, we PCR-amplified the exon spanning the site of this mutation from the genomic DNA of patients fulfilling DSM-III-R criteria for autistic disorder. None of the 119 patients tested were found to have this mutation. Furthermore, on preliminary screening using single-strand conformation polymorphism (SSCP), no novel mutations were detected in the coding sequence of four ADSL exons, spanning approximately 50% of the cDNA. In light of these findings, it appears that mutations in the ADSL gene represent a distinctly uncommon cause of autism. 12 refs., 2 figs.

  5. Evaluation of auxin and thiamine interaction effect on PAL activity and phenolic compounds content in vegetative growth stage of soybean plants

    Directory of Open Access Journals (Sweden)

    nazi nadernejad

    2017-08-01

    Full Text Available Soybean (Glycin max L. is one of the most important oily seeds in the world. This plant is rich in protein and unsaturated fats, and plays a significant role in human health with phenolic compounds and flavonoids. Indole Butyric Acid (IBA is a plant growth regulator that plays a key role in producing phenolic compounds and increasing the antioxidant capacity of the plant. Thiamine is one of the important vitamins in strengthening the immune system and increasing the resistance to environmental stresses in the plant's growth stages. Regarding the effect of hormone auxin and thiamine on the production of phenolic compounds as one of the antioxidant compounds in growth stages, the aim of this study was to investigate the effect of the two compounds in two stages of soybean growth and compare their effect on phenolic compounds changes in order to Detecting higher antioxidant capacity in environmental stress tolerance. For this purpose, the DPX cultivar of soybean seeds were prepared from Dezful Agriculture Research Center and planted in perlite containing flowers. The plants were planted under factorial design under IBA treatments with three concentrations of 0, 10 and 50 and thiamine with three concentrations of 0, 50 and 200. Extraction and evaluation of phenolic compounds, anthocyanins and pigments in leaves were performed. Data were analyzed using Duncan's test at a significant level of 5%. The results showed that the combined use of auxin and thiamine increased the carotenoid content in both phases and caused a significant increase in phenolic content. Application of auxin alone reduced auxin and thiamine the anthocyanin content significantly in both phases, but did not have a significant effect on phenolic content. The results showed that the PAL activity of the phenolic and anthocyanin content increased significantly in the 9-leaf stage compared to 3-leaf. Generally, the results showed that interaction effect between auxin and thiamine on

  6. A wild 'albino' bilberry (Vaccinium myrtillus L. from Slovenia shows three bottlenecks in the anthocyanin pathway and significant differences in the expression of several regulatory genes compared to the common blue berry type.

    Directory of Open Access Journals (Sweden)

    Zala Zorenc

    Full Text Available Relative expressions of structural genes and a number of transcription factors of the anthocyanin pathway relevant in Vaccinium species, and related key enzyme activities were compared with the composition and content of metabolites in skins of ripe fruits of wild albino and blue bilberry (Vaccinium myrtillus found in Slovenia. Compared to the common blue type, the albino variant had a 151-fold lower total anthocyanin and a 7-fold lower total phenolic content in their berry skin, which correlated with lower gene expression of flavonoid 3-O-glycosyltransferase (FGT; 33-fold, flavanone 3-hydroxylase (FHT; 18-fold, anthocyanidin synthase (ANS; 11-fold, chalcone synthase (CHS, 7.6-fold and MYBPA1 transcription factor (22-fold. The expression of chalcone isomerase (CHI, dihydroflavonol 4-reductase (DFR, leucoanthocyanidin reductase (LAR, anthocyanidin reductase (ANR and MYBC2 transcription factor was reduced only by a factor of 1.5-2 in the albino berry skins, while MYBR3 and flavonoid 3',5'-hydroxylase (F3'5'H were increased to a similar extent. Expression of the SQUAMOSA class transcription factor TDR4, in contrast, was independent of the color type and does therefore not seem to be correlated with anthocyanin formation in this variant. At the level of enzymes, significantly lower FHT and DFR activities, but not of phenylalanine ammonia-lyase (PAL and CHS/CHI, were observed in the fruit skins of albino bilberries. A strong increase in relative hydroxycinnamic acid derivative concentrations indicates the presence of an additional bottleneck in the general phenylpropanoid pathway at a so far unknown step between PAL and CHS.

  7. Hydrogen sulfide (H2S)/cystathionine γ-lyase (CSE) pathway contributes to the proliferation of hepatoma cells

    International Nuclear Information System (INIS)

    Pan, Yan; Ye, Shuang; Yuan, Dexiao; Zhang, Jianghong; Bai, Yang; Shao, Chunlin

    2014-01-01

    Highlights: • Inhibition of H 2 S/CSE pathway strongly stimulates cellular apoptosis. • Inhibition of H 2 S/CSE pathway suppresses cell growth by blocking EGFR pathway. • H 2 S/CSE pathway is critical for maintaining the proliferation of hepatoma cells. - Abstract: Hydrogen sulfide (H 2 S)/cystathionine γ-lyase (CSE) pathway has been demonstrated to play vital roles in physiology and pathophysiology. However, its role in tumor cell proliferation remains largely unclear. Here we found that CSE over-expressed in hepatoma HepG2 and PLC/PRF/5 cells. Inhibition of endogenous H 2 S/CSE pathway drastically decreased the proliferation of HepG2 and PLC/PRF/5 cells, and it also enhanced ROS production and mitochondrial disruption, pronounced DNA damage and increased apoptosis. Moreover, this increase of apoptosis was associated with the activation of p53 and p21 accompanied by a decreased ratio of Bcl-2/Bax and up-regulation of phosphorylated c-Jun N-terminal kinase (JNK) and caspase-3 activity. In addition, the negative regulation of cell proliferation by inhibition of H 2 S/CSE system correlated with the blockage of cell mitogenic and survival signal transduction of epidermal growth factor receptor (EGFR) via down-regulating the extracellular-signal-regulated kinase 1/2 (ERK1/2) activation. These results demonstrate that H 2 S/CSE and its downstream pathway contribute to the proliferation of hepatoma cells, and inhibition of this pathway strongly suppress the excessive growth of hepatoma cells by stimulating mitochondrial apoptosis and suppressing cell growth signal transduction

  8. Induction of resveratrol biosynthesis in skins of three grape cultivars by ultraviolet irradiation

    International Nuclear Information System (INIS)

    Takayanagi, Tsutomu; Okuda, Tohru; Mine, Yohei; Yokotsuka, Koki

    2004-01-01

    Resveratrol production and expression of the genes related to resveratrol biosynthesis were investigated in the skins of three Vitis vinifera cultivars Chardonnay, Koshu and an American hybrid grape, Muscat Bailey A (Bailey x Muscat Hamburg). Resveratrol concentration in the skins of all the grapes increased significantly when exposed to ultraviolet (UV-C, 254 nm) irradiation. The UV-induced resveratrol concentration in the grape skins was lower after veraison (onset of ripening) than before it. The maximum concentration of the UV-induced resveratrol in 'Muscat Bailey A' was higher than those in the other two cultivars. The relative mRNA expression levels of stilebene synthase (STS), phenylalanine ammonia-lyase (PAL) and chalcone synthase (CHS) genes in grape skins 8 hr after UV irradiation were determined by quantitative reverse transcription-polymerase chain reaction (RT-PCR). The results revealed that STS- and PAL-mRNA expressions were significantly increased by UV irradiation. STS-mRNA expressions in 'Muscat Bailey A' were higher than those in 'Chardonnay' throughout berry development. The UV-induced CHS-mRNA expression in the grape skins decreased before veraison and subsequently increased. (author)

  9. Effect of NaHCO3 treatments on the activity of cell wall-degrading enzymes produced by Penicillium digitatum during the pathogenesis process on grapefruit.

    Science.gov (United States)

    Venditti, Tullio; D'hallewin, Guy; Ladu, Gianfranca; Petretto, Giacomo L; Pintore, Giorgio; Labavitch, John M

    2018-03-25

    The present study was performed to clarify the strategies of Penicillium digitatum during pathogenesis on citrus, assessing, on albedo plugs, the effects of treatment with NaHCO 3 , at two different pH (5 and 8.3), on cell wall-degrading enzymes activity, over a period of 72 h. The treatment with NaHCO 3 , under alkaline pH, delayed the polygalacturonase activity for 72 h, or 48 h in the case of the pectin lyase, if compared to the control or the same treatment at pH 5. On the contrary, the pectin methyl esterase activity rapidly increased after 24 h, in plugs dipped in the same solution. In this case, the activity remained higher than untreated or pH 5 treated plugs up to 72 h. The rapid increase in pectin methyl esterase activity, under alkaline conditions, is presumably the strategy of the pathogen to lower the pH, soon after the initiation of infection, in order to restore an optimal environment for the subsequent polygalacturonase and pectin lyase action. In fact at the same time, a low pH delayed the enzymatic activity of polygalacturonase and pectin lyase, the two enzymes that actually cleave the α-1,4-linkages between the galacturonic acid residues. This article is protected by copyright. All rights reserved.

  10. Identifying Opportunities for Peer Learning: An Observational Study of Medical Students on Clinical Placements.

    Science.gov (United States)

    Tai, Joanna H; Canny, Benedict J; Haines, Terry P; Molloy, Elizabeth K

    2017-01-01

    Phenomenon: Peer assisted learning (PAL) is frequently employed and researched in preclinical medical education. Fewer studies have examined PAL in the clinical context: These have focused mainly on the accuracy of peer assessment and potential benefits to learner communication and teamwork skills. Research has also examined the positive and negative effects of formal, structured PAL activities in the clinical setting. Given the prevalence of PAL activities during preclinical years, and the unstructured nature of clinical placements, it is likely that nonformal PAL activities are also undertaken. How PAL happens formally and informally and why students find PAL useful in this clinical setting remain poorly understood. This study aimed to describe PAL activities within the context of clinical placement learning and to explore students' perceptions of these activities. An ethnographic study was conducted to gather empirical data on engagement in clinical placement learning activities, including observations and interviews with students in their 1st clinical year, along with their supervising clinicians. Thematic analysis was used to interrogate the data. On average, students used PAL for 5.19 hours per week in a range of activities, of a total of 29.29 hours undertaking placements. PAL was recognized as a means of vicarious learning and had greater perceived value when an educator was present to guide or moderate the learning. Trust between students was seen as a requirement for PAL to be effective. Students found passive observation a barrier to PAL and were able to identify ways to adopt an active stance when observing peers interacting with patients. For example, learners reported that the expectation that they had to provide feedback to peers after task observation, resulted in them taking on a more critical gaze where they were encouraged to consider notions of good practice. Insights: Students use PAL in formal (i.e., tutorial) and nonformal (e.g., peer

  11. Reliability and validity of two frequently used self-administered physical activity questionnaires in adolescents

    Directory of Open Access Journals (Sweden)

    Kurtze Nanna

    2008-07-01

    Full Text Available Abstract Background To create and find accurate and reliable instruments for the measurement of physical activity has been a challenge in epidemiological studies. We investigated the reliability and validity of two different physical activity questionnaires in 71 adolescents aged 13–18 years; the WHO, Health Behaviour in Schoolchildren (HBSC questionnaire, and the International Physical Activity Questionnaire (IPAQ, short version. Methods The questionnaires were administered twice (8–12 days apart to measure reliability. Validity was assessed by comparing answers from the questionnaires with a cardiorespiratory fitness test (VO2peak and seven days activity monitoring with the ActiReg, an instrument measuring physical activity level (PAL and total energy expenditure (TEE. Results Intraclass correlation coefficients for reliability for the WHO HBSC questionnaire were 0.71 for frequency and 0.73 for duration. For the frequency question, there was a significant difference between genders; 0.87 for girls and 0.59 for boys (p 2peak were fair, ranging between 0.29 – 0.39. The WHO HBSC questionnaire measured against VO2peak for girls were acceptable, ranging between 0.30 – 0.55. Both questionnaires, except the walking question in IPAQ, showed a low correlation with PAL and TEE, ranging between 0.01 and 0.29. Conclusion These data indicate that the WHO HBSC questionnaire had substantial reliability and were acceptable instrument for measuring cardiorespiratory fitness, especially among girls. None of the questionnaires however seemed to be a valid instrument for measuring physical activity compared to TEE and PAL in adolescents.

  12. A estrutura do palácio do Itamaraty em Brasília: Aspectos históricos e tecnológicos de projeto, execução, intervenções e manutenção

    OpenAIRE

    Santos, Evaristo; Clímaco, João; Nepomuceno, Antonio

    2007-01-01

    O Palácio do Itamaraty em Brasília é uma obra prima na Esplanada dos Ministérios da capital do Brasil, a única cidade construída no século XX laureada pela Unesco como Patrimônio Cultural da Humanidade, em 1989. O Palácio denota o gênio de seus criadores - o arquiteto Oscar Niemeyer e engenheiro estrutural Joaquim Cardozo, e a ousadia e qualidade dos profissionais de projeto e construção do Brasil. Baseado em documentos originais e depoimentos do pessoal envolvido na construção, entre 1963 e ...

  13. High Current, High Density Arc Plasma as a New Source for WiPAL

    Science.gov (United States)

    Waleffe, Roger; Endrizzi, Doug; Myers, Rachel; Wallace, John; Clark, Mike; Forest, Cary; WiPAL Team

    2016-10-01

    The Wisconsin Plasma Astrophysics Lab (WiPAL) has installed a new array of nineteen plasma sources (plasma guns) on its 3 m diameter, spherical vacuum vessel. Each gun is a cylindrical, molybdenum, washer-stabilized, arc plasma source. During discharge, the guns are maintained at 1.2 kA across 100 V for 10 ms by the gun power supply establishing a high density plasma. Each plasma source is fired independently allowing for adjustable plasma parameters, with densities varying between 1018 -1019 m-3 and electron temperatures of 5-15 eV. Measurements were characterized using a 16 tip Langmuir probe. The plasma source will be used as a background plasma for the magnetized coaxial plasma gun (MCPG), the Terrestrial Reconnection Experiment (TREX), and as the plasma source for a magnetic mirror experiment. Temperature, density, and confinement results will be presented. This work is supported by the DoE and the NSF.

  14. Physical activity level and sedentary behavior among students

    Directory of Open Access Journals (Sweden)

    Diego Augusto Santos Silva

    2009-01-01

    Full Text Available http://dx.doi.org/10.5007/1980-0037.2009v11n3p299   The objective of this study was to determine the physical activity level (PAL and sedentary behavior of students from the city of Aracaju (SE. A total of 1028 students of both genders participated in the study, with a mean age of 15.38 (2.44 years for girls and 15.24 (2.40 years for boys. Among the sample, 24.7% were children and 75.3% were adolescents, with a mean age of 12.07 (0.88 and 16.39 (1.72 years, respectively. An instrument already used in Brazilian studies was applied to identify the average time (hours watching TV per day (hTV and PAL – PAQ-C. Descriptive statistics, t-test for independent samples, Fisher’s exact test and comparison test between two proportions were used for data analysis, with the level of significance set at 5% (p≤0.05. Boys presented a significantly higher physical activity score 2.25 (0.60 than girls. The prevalence of sedentarism was 72.5, 89.3 and 85.2% in the groups of children and adolescents and in the group as a whole, respectively, for girls, and 55.4, 74.8 and 69.8% for boys. No differences in hTV were observed between genders or between sedentary and physically active students (p > 0.05. We conclude that a there is a high prevalence of “sedentary” and “very sedentary” children and adolescents; b boys present a higher PAL than girls; c adolescents are less active than children, and d the number of hTV is high in the group studied.

  15. Simultaneous measurement of two enzyme activities using infrared spectroscopy: A comparative evaluation of PARAFAC, TUCKER and N-PLS modeling

    DEFF Research Database (Denmark)

    Baum, Andreas; Hansen, Per Waaben; Meyer, Anne S.

    2013-01-01

    multiway methods, namely PARAFAC, TUCKER3 and N-PLS, to establish simultaneous enzyme activity assays for pectin lyase and pectin methyl esterase. Correlation coefficients Rpred2 for prediction test sets are 0.48, 0.96 and 0.96 for pectin lyase and 0.70, 0.89 and 0.89 for pectin methyl esterase......Enzymes are used in many processes to release fermentable sugars for green production of biofuel, or the refinery of biomass for extraction of functional food ingredients such as pectin or prebiotic oligosaccharides. The complex biomasses may, however, require a multitude of specific enzymes which...... are active on specific substrates generating a multitude of products. In this paper we use the plant polymer, pectin, to present a method to quantify enzyme activity of two pectolytic enzymes by monitoring their superimposed spectral evolutions simultaneously. The data is analyzed by three chemometric...

  16. Combined effects of Psoralens and ultraviolet-B on growth, pigmentation and biochemical parameters of Abelmoschus esculentus L.

    Science.gov (United States)

    Kumari, Rima; Singh, Suruchi; Agrawal, S B

    2009-05-01

    The effects of pre-treatment of Psoralens (furocoumarin compounds) and supplemental ultraviolet-B (sUV-B) were studied on plant growth, photosynthetic and non-photosynthetic pigments, protein, phenylalanine ammonia lyase (PAL) activity and antioxidative defense potential as well as their ultimate effects on biomass production in Abelmoschus esculentus L. (Okra) plants. Psoralens are capable of absorbing radiant energy and stimulating the pigmentation of human skin when photo-activated in presence of UV-A or UV-B making them beneficial in the treatment of vitilago. Pre-treatment of Psoralens against sUV-B (pUV-B), stimulates higher production of UV-B protective pigments (flavonoids and carotenoids) and helps in maintaining its biomass against UV-B stress. Antioxidative defense system in the test plant was activated by combined treatment of Psoralens and sUV-B as evidenced by the enhanced activity of enzymatic (ascorbate peroxidase-APX, superoxide dismutase-SOD, POX) and non-enzymatic (ascorbic acid and phenol) antioxidants. Individual treatments of Psoralens and sUV-B showed inhibitory effect on various morphological traits i.e. reduction in plant height, leaf area and ultimately on biomass production. Our results clearly indicated that adverse effect of sUV-B on biomass production was ameliorated by pre- treatment with Psoralens.

  17. Energy requirements and physical activity level of active elderly people in rural areas of cuba

    International Nuclear Information System (INIS)

    Hernandez-Triana, M.; Porrata Maury, C.; Jimenez Acosta, S.; Gonzalez Perez, T.; Diaz, M.E.; Martin, I.; Sanchez, V.; Monterrey, P.

    1999-01-01

    Obesity and non-insulin dependent diabetes mellitus (NIDDM) are common in the Third Age and increasing in Cuba. Among the life-style changes associated with increased prevalence of obesity and its related disorders, diet and activity patterns are prime candidates. The transition to this life-style model may induce a decrease in the energy needs. There is an urgent need for tools which have been validated for measuring diet and physical activity in nutritional studies in the developing world, but also a more urgent need for reference values for the total energy requirements of healthy elderly people. Regular physical activity reduces the likelihood to develop diseases that characterise the metabolic cardiovascular syndrome. Previous studies done in Havana showed values of physical activity level (PAL) which are lower than the reported for elderly subjects. Elderly people living in rural areas use to have physical activity levels which differ from the observed in urban areas. With the purpose of estimating the energy requirements, a group of 40 apparently healthy people older than 60 years of age living in a rural mountain community will be submitted to a medical, epidemiological, dietary, anthropometric and insulin resistance study. Physical activity will be determined by questionnaire and by the calculation of the PAL from the basal metabolic rate (BMR) and total energy expenditure (TEE) measured with the doubly-labelled water method (DLW). Associations with the prevalence of insulin resistance and obesity will be assessed. (author)

  18. Fungal Endophytes as a Metabolic Fine-Tuning Regulator for Wine Grape.

    Directory of Open Access Journals (Sweden)

    Ming-Zhi Yang

    Full Text Available Endophytes proved to exert multiple effects on host plants, including growth promotion, stress resistance. However, whether endophytes have a role in metabolites shaping of grape has not been fully understood. Eight endophytic fungal strains which originally isolated from grapevines were re-inoculated to field-grown grapevines in this study, and their effects on both leaves and berries of grapevines at maturity stage were assessed, with special focused on secondary metabolites and antioxidant activities. High-density inoculation of all these endophytic fungal strains modified the physio-chemical status of grapevine to different degrees. Fungal inoculations promoted the content of reducing sugar (RS, total flavonoids (TF, total phenols (TPh, trans-resveratrol (Res and activities of phenylalanine ammonia-lyase (PAL, in both leaves and berries of grapevine. Inoculation of endophytic fungal strains, CXB-11 (Nigrospora sp. and CXC-13 (Fusarium sp. conferred greater promotion effects in grape metabolic re-shaping, compared to other used fungal strains. Additionally, inoculation of different strains of fungal endophytes led to establish different metabolites patterns of wine grape. The work implies the possibility of using endophytic fungi as fine-tuning regulator to shape the quality and character of wine grape.

  19. Cystathionine γ-Lyase-Produced Hydrogen Sulfide Controls Endothelial NO Bioavailability and Blood Pressure.

    Science.gov (United States)

    Szijártó, István András; Markó, Lajos; Filipovic, Milos R; Miljkovic, Jan Lj; Tabeling, Christoph; Tsvetkov, Dmitry; Wang, Ning; Rabelo, Luiza A; Witzenrath, Martin; Diedrich, André; Tank, Jens; Akahoshi, Noriyuki; Kamata, Shotaro; Ishii, Isao; Gollasch, Maik

    2018-06-01

    Hydrogen sulfide (H 2 S) and NO are important gasotransmitters, but how endogenous H 2 S affects the circulatory system has remained incompletely understood. Here, we show that CTH or CSE (cystathionine γ-lyase)-produced H 2 S scavenges vascular NO and controls its endogenous levels in peripheral arteries, which contribute to blood pressure regulation. Furthermore, eNOS (endothelial NO synthase) and phospho-eNOS protein levels were unaffected, but levels of nitroxyl were low in CTH-deficient arteries, demonstrating reduced direct chemical interaction between H 2 S and NO. Pretreatment of arterial rings from CTH-deficient mice with exogenous H 2 S donor rescued the endothelial vasorelaxant response and decreased tissue NO levels. Our discovery that CTH-produced H 2 S inhibits endogenous endothelial NO bioavailability and vascular tone is novel and fundamentally important for understanding how regulation of vascular tone is tailored for endogenous H 2 S to contribute to systemic blood pressure function. © 2018 American Heart Association, Inc.

  20. Thermodependence of growth and enzymatic activities implicated in pathogenicity of two Erwinia carotovora subspecies (Pectobacterium spp.).

    Science.gov (United States)

    Smadja, Bruno; Latour, Xavier; Trigui, Sameh; Burini, Jean François; Chevalier, Sylvie; Orange, Nicole

    2004-01-01

    Erwinia carotovora subsp. atroseptica and Erwinia carotovora subsp. carotovora can cause substantial damage to economically important plant crops and stored products. The occurrence of the disease and the scale of the damage are temperature dependent. Disease development consists first of active multiplication of the bacteria in the infection area and then production of numerous extracellular enzymes. We investigated the effects of various temperatures on these two steps. We assayed the specific growth rate and the pectate lyase and protease activities for eight strains belonging to E. carotovora subsp. atroseptica and E. carotovora subsp. carotovora in vitro. The temperature effect on growth rate and on pectate lyase activity is different for the two subspecies, but protease activity appears to be similarly thermoregulated. Our results are in agreement with ecological data implicating E. carotovora subsp. atroseptica in disease when the temperature is below 20 degrees C. The optimal temperature for pathogenicity appears to be different from the optimal growth temperature but seems to be a compromise between this temperature and temperatures at which lytic activities are maximal.

  1. Basic features of electromagnetic pulse generated in a laser-target chamber at 3-TW laser facility PALS

    International Nuclear Information System (INIS)

    De Marco, M; Pfeifer, M; Krousky, E; Krasa, J; Cikhardt, J; Klir, D; Nassisi, V

    2014-01-01

    We describe the radiofrequency emission taking place when 300 ps laser pulses irradiate various solid targets with an intensity of 10 16 W/cm 2 . The emission of intense electromagnetic pulses was observed outside the laser target chamber by two loop antennas up to 1 GHz. Electromagnetic pulses can be 800 MHz transients, which decay from a peak electromagnetic field of E 0 ≊ 7 kV/m and H 0 ≊ 15 A/m. The occurrence of these electromagnetic pulses is associated with generation of hard x-rays with photon energies extending beyond 1 MeV. This contribution reports the first observation of this effect at the PALS facility.

  2. Understanding the role of argininosuccinate lyase transcript variants in the clinical and biochemical variability of the urea cycle disorder argininosuccinic aciduria.

    Science.gov (United States)

    Hu, Liyan; Pandey, Amit V; Eggimann, Sandra; Rüfenacht, Véronique; Möslinger, Dorothea; Nuoffer, Jean-Marc; Häberle, Johannes

    2013-11-29

    Argininosuccinic aciduria (ASA) is an autosomal recessive urea cycle disorder caused by deficiency of argininosuccinate lyase (ASL) with a wide clinical spectrum from asymptomatic to severe hyperammonemic neonatal onset life-threatening courses. We investigated the role of ASL transcript variants in the clinical and biochemical variability of ASA. Recombinant proteins for ASL wild type, mutant p.E189G, and the frequently occurring transcript variants with exon 2 or 7 deletions were (co-)expressed in human embryonic kidney 293T cells. We found that exon 2-deleted ASL forms a stable truncated protein with no relevant activity but a dose-dependent dominant negative effect on enzymatic activity after co-expression with wild type or mutant ASL, whereas exon 7-deleted ASL is unstable but seems to have, nevertheless, a dominant negative effect on mutant ASL. These findings were supported by structural modeling predictions for ASL heterotetramer/homotetramer formation. Illustrating the physiological relevance, the predominant occurrence of exon 7-deleted ASL was found in two patients who were both heterozygous for the ASL mutant p.E189G. Our results suggest that ASL transcripts can contribute to the highly variable phenotype in ASA patients if expressed at high levels. Especially, the exon 2-deleted ASL variant may form a heterotetramer with wild type or mutant ASL, causing markedly reduced ASL activity.

  3. Changes in antioxidant and biochemical activities in castor oil-coated Capsicum annuum L. during postharvest storage.

    Science.gov (United States)

    Panigrahi, Jitendriya; Patel, Mansi; Patel, Niyati; Gheewala, Bhumi; Gantait, Saikat

    2018-06-01

    This study, for the first time, evaluates the efficiency of castor oil when used as an external coating on Capsicum annuum L., to increase postharvest storage-life at 4 ± 1 °C. The castor oil-coated fruits were successfully stored for 36 days, while the non-coated fruits could only sustain for 18 days. Throughout the storage period (at 9-day intervals), different antioxidants and biochemical assays (allied with storage) such as titratable acidity, ascorbic acid content, ferrous ion chelating activity, reducing power, DPPH scavenging activity, hydroxyl radical scavenging activity, total phenolic content, total sugar estimation, and enzymatic study of polyphenol oxidase and pectate lyase, were assessed. During storage, the castor oil-coated fruits showed a substantial decrease in titratable acidity, ascorbic acid content, total phenolic content, including antioxidant activities such as reducing power and DPPH activity; however, an increase in ferrous ion chelating activity, total soluble sugar content, polyphenol oxidase activity and initial pectate lyase activity was observed, in contrast to that of the non-coated fruits. The application of castor oil proved to be effective in delaying the ripening process of fruits during storage.

  4. Plant defense induced in in vitro propagated banana (Musa paradisiaca) plantlets by Fusarium derived elicitors.

    Science.gov (United States)

    Patel, Miral; Kothari, I L; Mohan, J S S

    2004-07-01

    Perception of microbial signal molecules is part of the strategy evolved by plants to survive attacks by potential pathogens. To gain a more complete understanding of the early signaling events involved in these responses, we used fungal components of Fusarium under in vitro condition and checked the rise in signal molecule, salicylic acid (SA), and marker enzymes in defense reactions against the pathogen. SA level increased by 21 folds in elicitor treated plantlets as compared to that of control plantlets and there was marked increase in phenylalanine ammonia-lyase(PAL), peroxidase(POX), polyphenol oxidase(PPO) along with higher total phenolic content. Present results indicated that use of fungal components had successfully induced systemic resistance in in vitro cultured banana plantlets.

  5. Sublethal Heavy Metal Stress Stimulates Innate Immunity in Tomato

    Directory of Open Access Journals (Sweden)

    Nilanjan Chakraborty

    2015-01-01

    Full Text Available Effect of sublethal heavy metal stress as plant biotic elicitor for triggering innate immunity in tomato plant was investigated. Copper in in vivo condition induced accumulation of defense enzymes like peroxidase (PO, polyphenol oxidase (PPO, phenylalanine ammonia-lyase (PAL, and β-1,3 glucanase along with higher accumulation of total phenol, antioxidative enzymes (catalase and ascorbate peroxidase, and total chlorophyll content. Furthermore, the treatment also induced nitric oxide (NO production which was confirmed by realtime visualization of NO burst using a fluorescent probe 4,5-diaminofluorescein diacetate (DAF-2DA and spectrophotometric analysis. The result suggested that the sublethal dose of heavy metal can induce an array of plant defense responses that lead to the improvement of innate immunity in plants.

  6. Mitos e Lendas como contributo para a diferenciação da Experiência Turística – o caso dos Túneis do Palácio Nacional de Mafra, Portugal

    Directory of Open Access Journals (Sweden)

    Andreia Sofia Costa Morgado

    2016-10-01

    Full Text Available Desmitificar os Túneis do Palácio Nacional de Mafra, Portugal, perspetivando novas formas de diferenciação de uma oferta turística associada a um imaginário de exotismo e mistério, foi o principal propósito do presente trabalho. Os mitos relativos ao Palácio Nacional de Mafra integram a cultura popular mafrense, pelo que se coloca a possibilidade de envolver a comunidade local com base na apropriação simbólica da sua realidade. O pressuposto do envolvimento comunitário local foi equacionado nos inquéritos realizados junto da comunidade local e, assim, as ofertas turísticas alternativas assumem-se como uma oportunidade de diferenciação da Experiência Turística em Mafra.

  7. A QM/MM study of the reaction mechanism of (R)-hydroxynitrile lyases from Arabidopsis thaliana (AtHNL).

    Science.gov (United States)

    Zhu, Wenyou; Liu, Yongjun; Zhang, Rui

    2015-01-01

    Hydroxynitrile lyases (HNLs) catalyze the conversion of chiral cyanohydrins to hydrocyanic acid (HCN) and aldehyde or ketone. Hydroxynitrile lyase from Arabidopsis thaliana (AtHNL) is the first R-selective HNL enzyme containing an α/β-hydrolases fold. In this article, the catalytic mechanism of AtHNL was theoretically studied by using QM/MM approach based on the recently obtained crystal structure in 2012. Two computational models were constructed, and two possible reaction pathways were considered. In Path A, the calculation results indicate that the proton transfer from the hydroxyl group of cyanohydrin occurs firstly, and then the cleavage of C1-C2 bond and the rotation of the generated cyanide ion (CN(-)) follow, afterwards, CN(-) abstracts a proton from His236 via Ser81. The C1-C2 bond cleavage and the protonation of CN(-) correspond to comparable free energy barriers (12.1 vs. 12.2 kcal mol(-1)), suggesting that both of the two processes contribute a lot to rate-limiting. In Path B, the deprotonation of the hydroxyl group of cyanohydrin and the cleavage of C1-C2 bond take place in a concerted manner, which corresponds to the highest free energy barrier of 13.2 kcal mol(-1). The free energy barriers of Path A and B are very similar and basically agree well with the experimental value of HbHNL, a similar enzyme of AtHNL. Therefore, both of the two pathways are possible. In the reaction, the catalytic triad (His236, Ser81, and Asp208) acts as the general acid/base, and the generated CN(-) is stabilized by the hydroxyl group of Ser81 and the main-chain NH-groups of Ala13 and Phe82. © 2014 Wiley Periodicals, Inc.

  8. Isolation of a novel alginate lyase-producing Bacillus litoralis strain and its potential to ferment Sargassum horneri for biofertilizer.

    Science.gov (United States)

    Wang, Mingpeng; Chen, Lei; Liu, Zhengyi; Zhang, Zhaojie; Qin, Song; Yan, Peisheng

    2016-12-01

    Algae have long been used to augment plant productivity through their beneficial effects. Alginate oligosaccharide is believed to be one of the important components to enhance growth and crop yield. In this study, we isolated and characterized a Bacillus litoralis strain, named Bacillus M3, from decayed kelps. We further demonstrated that the M3 strain could secrete alginate lyase to degrade alginate. The crude enzyme exhibited the highest activity (33.74 U/mg) at pH 7.0 and 50°C. The M3 strain was also able to ferment the brown alga Sargassum horneri. Fermentation results revealed that a fermentation period of 8-12 hr was the best harvest time with the highest level of alginate oligosaccharides. Plant growth assay showed that the seaweed fermentation extract had an obvious promotion effect on root and seedling growth of Lycopersicon eseulentum L. Our results suggest that fermentation extract of Sargassum horneri by the novel strain of Bacillus litoralis M3 has significant development potential for biofertilizer production and agriculture application. © 2016 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd.

  9. RegA Plays a Key Role in Oxygen-Dependent Establishment of Persistence and in Isocitrate Lyase Activity, a Critical Determinant of In vivo Brucella suis Pathogenicity

    Directory of Open Access Journals (Sweden)

    Elias Abdou

    2017-05-01

    Full Text Available For aerobic human pathogens, adaptation to hypoxia is a critical factor for the establishment of persistent infections, as oxygen availability is low inside the host. The two-component system RegB/A of Brucella suis plays a central role in the control of respiratory systems adapted to oxygen deficiency, and in persistence in vivo. Using an original “in vitro model of persistence” consisting in gradual oxygen depletion, we compared transcriptomes and proteomes of wild-type and ΔregA strains to identify the RegA-regulon potentially involved in the set-up of persistence. Consecutive to oxygen consumption resulting in growth arrest, 12% of the genes in B. suis were potentially controlled directly or indirectly by RegA, among which numerous transcriptional regulators were up-regulated. In contrast, genes or proteins involved in envelope biogenesis and in cellular division were repressed, suggesting a possible role for RegA in the set-up of a non-proliferative persistence state. Importantly, the greatest number of the RegA-repressed genes and proteins, including aceA encoding the functional IsoCitrate Lyase (ICL, were involved in energy production. A potential consequence of this RegA impact may be the slowing-down of the central metabolism as B. suis progressively enters into persistence. Moreover, ICL is an essential determinant of pathogenesis and long-term interactions with the host, as demonstrated by the strict dependence of B. suis on ICL activity for multiplication and persistence during in vivo infection. RegA regulates gene or protein expression of all functional groups, which is why RegA is a key regulator of B. suis in adaptation to oxygen depletion. This function may contribute to the constraint of bacterial growth, typical of chronic infection. Oxygen-dependent activation of two-component systems that control persistence regulons, shared by several aerobic human pathogens, has not been studied in Brucella sp. before. This work

  10. Purification, molecular cloning, and expression of 2-hydroxyphytanoyl- CoA lyase, a peroxisomal thiamine pyrophosphate-dependent enzyme that catalyzes the carbon-carbon bond cleavage during à-oxidation of 3- methyl-branched fatty acids

    CERN Document Server

    Foulon, V; Croes, K; Waelkens, E

    1999-01-01

    Purification, molecular cloning, and expression of 2-hydroxyphytanoyl- CoA lyase, a peroxisomal thiamine pyrophosphate-dependent enzyme that catalyzes the carbon-carbon bond cleavage during à-oxidation of 3- methyl-branched fatty acids

  11. Solvent Isotope-induced Equilibrium Perturbation for Isocitrate Lyase

    Science.gov (United States)

    Quartararo, Christine E.; Hadi, Timin; Cahill, Sean M.; Blanchard, John S.

    2014-01-01

    Isocitrate lyase (ICL) catalyzes the reversible retro-aldol cleavage of isocitrate to generate glyoxylate and succinate. ICL is the first enzyme of the glyoxylate shunt, which allows for the anaplerosis of citric acid cycle intermediates under nutrient limiting conditions. In Mycobacterium tuberculosis, the source of ICL for these studies, ICL is vital for the persistence phase of the bacteria’s life cycle. Solvent kinetic isotope effects (KIEs) in the direction of isocitrate cleavage of D2OV = 2.0 ± 0.1 and D2O[V/Kisocitrate] = 2.2 ± 0.3 arise from the initial deprotonation of the C2 hydroxyl group of isocitrate or the protonation of the aci-acid of succinate product of the isocitrate aldol cleavage by a solvent-derived proton. This KIE suggested that an equilibrium mixture of all protiated isocitrate, glyoxylate and succinate prepared in D2O, would undergo transient changes in equilibrium concentrations as a result of the solvent KIE and solvent-derived deuterium incorporation into both succinate and isocitrate. No change in the isotopic composition of glyoxylate was expected or observed. We have directly monitored the changing concentrations of all isotopic species of all reactants and products using a combination of NMR spectroscopy and mass spectrometry. Continuous monitoring of glyoxylate by 1H NMR spectroscopy shows a clear equilibrium perturbation in D2O. The final equilibrium isotopic composition of reactants in D2O revealed di-deuterated succinate, protiated glyoxylate, and mono-deuterated isocitrate, with the transient appearance and disappearance of mono-deuterated succinate. A model for the equilibrium perturbation of substrate species, and their time-dependent isotopic composition is presented. PMID:24261638

  12. Functional Analyses of Resurrected and Contemporary Enzymes Illuminate an Evolutionary Path for the Emergence of Exolysis in Polysaccharide Lyase Family 2.

    Science.gov (United States)

    McLean, Richard; Hobbs, Joanne K; Suits, Michael D; Tuomivaara, Sami T; Jones, Darryl R; Boraston, Alisdair B; Abbott, D Wade

    2015-08-28

    Family 2 polysaccharide lyases (PL2s) preferentially catalyze the β-elimination of homogalacturonan using transition metals as catalytic cofactors. PL2 is divided into two subfamilies that have been generally associated with secretion, Mg(2+) dependence, and endolysis (subfamily 1) and with intracellular localization, Mn(2+) dependence, and exolysis (subfamily 2). When present within a genome, PL2 genes are typically found as tandem copies, which suggests that they provide complementary activities at different stages along a catabolic cascade. This relationship most likely evolved by gene duplication and functional divergence (i.e. neofunctionalization). Although the molecular basis of subfamily 1 endolytic activity is understood, the adaptations within the active site of subfamily 2 enzymes that contribute to exolysis have not been determined. In order to investigate this relationship, we have conducted a comparative enzymatic analysis of enzymes dispersed within the PL2 phylogenetic tree and elucidated the structure of VvPL2 from Vibrio vulnificus YJ016, which represents a transitional member between subfamiles 1 and 2. In addition, we have used ancestral sequence reconstruction to functionally investigate the segregated evolutionary history of PL2 progenitor enzymes and illuminate the molecular evolution of exolysis. This study highlights that ancestral sequence reconstruction in combination with the comparative analysis of contemporary and resurrected enzymes holds promise for elucidating the origins and activities of other carbohydrate active enzyme families and the biological significance of cryptic metabolic pathways, such as pectinolysis within the zoonotic marine pathogen V. vulnificus. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

  13. The Richmond Agitation-Sedation Scale modified for palliative care inpatients (RASS-PAL): a pilot study exploring validity and feasibility in clinical practice

    OpenAIRE

    Bush, Shirley H; Grassau, Pamela A; Yarmo, Michelle N; Zhang, Tinghua; Zinkie, Samantha J; Pereira, José L

    2014-01-01

    Background The Richmond Agitation-Sedation Scale (RASS), which assesses level of sedation and agitation, is a simple observational instrument which was developed and validated for the intensive care setting. Although used and recommended in palliative care settings, further validation is required in this patient population. The aim of this study was to explore the validity and feasibility of a version of the RASS modified for palliative care populations (RASS-PAL). Methods A prospective study...

  14. Phosphorylation of sites 3 and 2 in rabbit skeletal muscle glycogen synthase by a multifunctional protein kinase (ATP-citrate lyase kinase)

    International Nuclear Information System (INIS)

    Sheorain, V.S.; Ramakrishna, S.; Benjamin, W.B.; Soderling, T.R.

    1985-01-01

    A multifunctional protein kinase, purified from rat liver as ATP-citrate lyase kinase, has been identified as a glycogen synthase kinase. This kinase catalyzed incorporation of up to 1.5 mol of and]2number 2 PO 4 /mol of synthase subunit associated with a decrease in the glycogen synthase activity ratio from 0.85 to a value of 0.15. Approximately 65-70% of the 34 PO 4 was incorporated into site 3 and 30-35% into site 2 as determined by reverse phase high performance liquid chromatography. This multifunctional kinase was distinguished from glycogen synthase kinase-3 on the basis of nucleotide and protein substrate specificities. Since the phosphate contents in glycogen synthase of sites 3 and 2 are altered in diabetes and by insulin administration, the possible involvement of the multifunctional kinase was explored. Glycogen synthase purified from diabetic rabbits was phosphorylated in vitro by this multifunctional kinase at only 10% of the rate compared to synthase purified from control rabbits. Treatment of the diabetics with insulin restored the synthase to a form that was readily phosphorylated in vitro

  15. Comparison of the free volume sizes and shapes determined from crystallographic and PALS data

    Directory of Open Access Journals (Sweden)

    Tydda Maciej

    2015-12-01

    Full Text Available Two different classes of molecular crystals were investigated. The first group was benzenediols, which are characterized by the same chemical composition but a different organization of their crystallographic structures; all of the compounds from this group have only one kind of free volumes. The second class was represented by olanzapine, which has more complex chemical composition and two kinds of free volumes in the structure. The o-Ps lifetime values determined from positron annihilation lifetime spectroscopy (PALS measurements agree quite well with those calculated for sizes found from crystallographic data for benzenediols (agreement within 10% of the lifetime values. For olanzapine, a good agreement is observed in the case of cuboidal free volumes, while for the other kind of void, the agreement is less satisfactory. Positronium diffusion coefficient determined from o-Ps redistribution in olanzapine agrees with these found for polymers.

  16. Partial deficiency of sphingosine-1-phosphate lyase confers protection in experimental autoimmune encephalomyelitis.

    Directory of Open Access Journals (Sweden)

    Andreas Billich

    Full Text Available BACKGROUND: Sphingosine-1-phosphate (S1P regulates the egress of T cells from lymphoid organs; levels of S1P in the tissues are controlled by S1P lyase (Sgpl1. Hence, Sgpl1 offers a target to block T cell-dependent inflammatory processes. However, the involvement of Sgpl1 in models of disease has not been fully elucidated yet, since Sgpl1 KO mice have a short life-span. METHODOLOGY: We generated inducible Sgpl1 KO mice featuring partial reduction of Sgpl1 activity and analyzed them with respect to sphingolipid levels, T-cell distribution, and response in models of inflammation. PRINCIPAL FINDINGS: The partially Sgpl1 deficient mice are viable but feature profound reduction of peripheral T cells, similar to the constitutive KO mice. While thymic T cell development in these mice appears normal, mature T cells are retained in thymus and lymph nodes, leading to reduced T cell numbers in spleen and blood, with a skewing towards increased proportions of memory T cells and T regulatory cells. The therapeutic relevance of Sgpl1 is demonstrated by the fact that the inducible KO mice are protected in experimental autoimmune encephalomyelitis (EAE. T cell immigration into the CNS was found to be profoundly reduced. Since S1P levels in the brain of the animals are unchanged, we conclude that protection in EAE is due to the peripheral effect on T cells, leading to reduced CNS immigration, rather than on local effects in the CNS. SIGNIFICANCE: The data suggest Sgpl1 as a novel therapeutic target for the treatment of multiple sclerosis.

  17. Seaweed Polysaccharides and Derived Oligosaccharides Stimulate Defense Responses and Protection Against Pathogens in Plants

    Directory of Open Access Journals (Sweden)

    Alejandra Moenne

    2011-11-01

    Full Text Available Plants interact with the environment by sensing “non-self” molecules called elicitors derived from pathogens or other sources. These molecules bind to specific receptors located in the plasma membrane and trigger defense responses leading to protection against pathogens. In particular, it has been shown that cell wall and storage polysaccharides from green, brown and red seaweeds (marine macroalgae corresponding to ulvans, alginates, fucans, laminarin and carrageenans can trigger defense responses in plants enhancing protection against pathogens. In addition, oligosaccharides obtained by depolymerization of seaweed polysaccharides also induce protection against viral, fungal and bacterial infections in plants. In particular, most seaweed polysaccharides and derived oligosaccharides trigger an initial oxidative burst at local level and the activation of salicylic (SA, jasmonic acid (JA and/or ethylene signaling pathways at systemic level. The activation of these signaling pathways leads to an increased expression of genes encoding: (i Pathogenesis-Related (PR proteins with antifungal and antibacterial activities; (ii defense enzymes such as pheylalanine ammonia lyase (PAL and lipoxygenase (LOX which determine accumulation of phenylpropanoid compounds (PPCs and oxylipins with antiviral, antifugal and antibacterial activities and iii enzymes involved in synthesis of terpenes, terpenoids and/or alkaloids having antimicrobial activities. Thus, seaweed polysaccharides and their derived oligosaccharides induced the accumulation of proteins and compounds with antimicrobial activities that determine, at least in part, the enhanced protection against pathogens in plants.

  18. Hyperhydricity in micropropagated carnation shoots: the role of oxidative stress.

    Science.gov (United States)

    Saher, Shady; Piqueras, Abel; Hellin, Eladio; Olmos, Enrique

    2004-01-01

    The physiology of hyperhydricity in relation to oxidative stress, mineral nutrients, antioxidant enzymes and ethylene has been studied in three micropropagated carnation cultivars under experimentally induced hyperhydricity. A marked increase in Fe content in comparison with normal tissues was observed in the hyperhydric tissues from the three cultivars. The levels of ethylene, solute leakage and malondialdehyde content were also significantly higher in the hyperhydric tissues. In relation to the time course of H(2)O(2) production measured by fluorescence quenching, a similar trend could be observed for the three cultivars, with a clear increase in the generation of hydrogen peroxide in hyperhydric tissues. The activities of all the antioxidative enzymes studied, except lipoxygenase, were higher in the hyperhydric shoots. Phenylalanine ammonia-lyase (PAL) showed a significant decrease in activity in the hyperhydric tissues in comparison with the controls for the three cultivars. Soluble guaiacol peroxidase had a strong increase in activity in hyperhydric shoots of the three cultivars. These results provide, for the first time, direct evidence of H(2)O(2) generation in hyperhydric tissues, characterize the response of the antioxidant system to an oxidative stress during hyperhydricity in carnation leaves and point to the accumulation of toxic forms of oxygen as the inducer of some of the abnormalities observed.

  19. Measurement of Sedentary Behaviors or "Downtime" in Rett Syndrome

    DEFF Research Database (Denmark)

    Stahlhut, Michelle; Hill, Kylie; Bisgaard, Anne-Marie

    2017-01-01

    measured duration of sedentary time with a mean difference (limit of agreement) of -1.0 (6.3) minutes. The duration of Bouchard activity record downtime accounted for 73% of the variance of sedentary time measured by the activPAL (coefficient 0.762, 95% CI 0.413 to 1.111). These data provide clinicians......This study aimed to validate measures of sedentary time in individuals with Rett syndrome. Twenty-six individuals (median [IQR] age 16.0 (9.4-20.6) years) wore an activPAL accelerometer during video-taped activities and agreement was determined between sedentary time determined by the activ......PAL and observation. For 11 individuals (median [IQR] age 14.5 (11.5-25.6) years), linear regression was used to determine the relationship between sedentary time recorded on the modified Bouchard activity record diary card and measured using the activPAL. In comparison to observation, the activPAL accurately...

  20. Trichoderma harzianum elicits induced resistance in sunflower challenged by Rhizoctonia solani.

    Science.gov (United States)

    Singh, B N; Singh, A; Singh, B R; Singh, H B

    2014-03-01

    To investigate the efficacy of Trichoderma harzianum NBRI-1055 (denoted as 'T-1055') in suppression of seedling blight of sunflower caused by Rhizoctonia solani Kühn and their impact on host defence responses. T-1055 was applied as seed treatment, soil application and combined application (seed treatment + soil application). Higher protection afforded by combined application of T-1055 was associated with the marked induction of phenylalanine ammonia-lyase (PAL), polyphenol oxidase (PPO), peroxidase (PO) and cinnamyl alcohol dehydrogenase (CAD) activities. The activities of PAL and PPO reached maximum at 10 days after sowing (DAS), while PO and CAD levels reached maximum at 12 DAS. This was further supported by the accumulation of total phenolic content that showed an increase up to threefold at 14 DAS. In addition, HPLC analysis revealed that the contents of ferulic and p-coumaric acids increased by 6·3 and 4·6 times, respectively, at 14 DAS. Amount of gallic acid was also little more than double. Lignin deposition in sunflower root increased by 2·7, 3·4 and 3·7 times through combined application of T-1055 at 16, 18 and 20 DAS, respectively. Combined application also increased the accumulation of PR-2 and PR-3 proteins by 3·3 and 3·9 times, respectively, at 12 DAS in followed by seed treatment alone. The combined application of T-1055 triggered defence responses in an enhanced level in sunflower than the soil and seed alone and provided better protection against Rhizoctonia seedling blight. Rhizospheric fungal bioagent 'T-1055' can enhance protection in sunflower against the R. solani pathogen through augmented elicitation of host defence responses. © 2013 The Society for Applied Microbiology.

  1. Electromagnetic Field Effects on Production of Salicylic Acid and Phenylalanine ammonia lyase in Tobacco Infected with Potato Virus X (PVX

    Directory of Open Access Journals (Sweden)

    amin radmard titkanlo

    2018-02-01

    . If the treatment number 11, OD (2/85 and in pollution-free treatment that was an OD (0/35 was observed. The OD also in other treatments were not exposed between 2.00 to 2.46 were visible. The results showed that the treatment 11 (infected control actually infected treatment that has not been exposed, and treatment 8 is the most desirable treatment for a reduction in the OD, the difference in the OD are available 0/98. The results suggest that the electromagnetic field decreases the amount of OD in tobacco seedlings are infected with PVX. Researchers said electromagnetic field growth rate and the number of spores grown in three species are affected. In another study, it was found that the impact on dry and wet mycorrhiza. The electromagnetic field decreases witches' broom disease infected lemon trees that were probably due to the decline phytoplasmas in plant tissues. Another study on the antibacterial effects of electromagnetic fields on Serratia marcescens was conducted, which resulted in the inhibition of bacterial growth. The results of salicylic acid and PAL determine the tobacco crop treatments, there are significant differences in the level of 0/05. So that treatment (7 (50 mT, 60 min the highest salicylic acid and treatment 1 (control non-infected showed the lowest rates. Also, reviews PAL treatment and treatment (7 most (1 (see non-polluting had the lowest. In fact, with the arrival of the pathogen to the host salicylic acid and phenylalanine ammonia will increase. But the electromagnetic stress in some treatments of both traits, from infected plants, should take over. However, some treatments less than 11 treatments (infection control is (but treatment Salicylic acid (4 was less than healthy plant. Interestingly, in both treatments was the highest for both traits in treatments of 7, 6 and 5, respectively. Pre-plant treatments with salicylic acid caused a significant increase in the PAL activity, which indicates the role of these enzymes in plant defense

  2. Influence of gallic and tannic acids on enzymatic activity and growth ...

    African Journals Online (AJOL)

    The effect of phenolic acids (gallic and tannic acids) on growth of Pectobacterium chrysanhemi, and its protease and pectate lyase activities was tested. The results obtained showed a significant inhibiting effect of the tannic and gallic acids on the growth of this strain. The growth rate decreases in the presence of 400 g/ml ...

  3. Bioproduction of L-Aspartic Acid and Cinnamic Acid by L-Aspartate Ammonia Lyase from Pseudomonas aeruginosa PAO1.

    Science.gov (United States)

    Patel, Arti T; Akhani, Rekha C; Patel, Manisha J; Dedania, Samir R; Patel, Darshan H

    2017-06-01

    Aspartase (L-aspartate ammonia lyase, EC 4.3.1.1) catalyses the reversible amination and deamination of L-aspartic acid to fumaric acid which can be used to produce important biochemical. In this study, we have explored the characteristics of aspartase from Pseudomonas aeruginosa PAO1 (PA-AspA). To overproduce PA-AspA, the 1425-bp gene was introduced in Escherichia coli BL21 and purified. A 51.0-kDa protein was observed as a homogenous purified protein on SDS-PAGE. The enzyme was optimally active at pH 8.0 and 35 °C. PA-AspA has retained 56% activity after 7 days of incubation at 35 °C, which displays the hyperthermostablility characteristics of the enzyme. PA-AspA is activated in the presence of metal ions and Mg2+ is found to be most effective. Among the substrates tested for specificity of PA-AspA, L-phenylalanine (38.35 ± 2.68) showed the highest specific activity followed by L-aspartic acid (31.21 ± 3.31) and fumarate (5.42 ± 2.94). K m values for L-phenylalanine, L-aspartic acid and fumarate were 1.71 mM, 0.346 μM and 2 M, respectively. The catalytic efficiency (k cat /K m ) for L-aspartic acid (14.18 s -1  mM -1 ) was higher than that for L-phenylalanine (4.65 s -1  mM -1 ). For bioconversion, from an initial concentration of 1000 mM of fumarate and 30 mM of L-phenylalanine, PA-AspA was found to convert 395.31 μM L-aspartic acid and 3.47 mM cinnamic acid, respectively.

  4. Hydrogen sulfide (H{sub 2}S)/cystathionine γ-lyase (CSE) pathway contributes to the proliferation of hepatoma cells

    Energy Technology Data Exchange (ETDEWEB)

    Pan, Yan; Ye, Shuang; Yuan, Dexiao; Zhang, Jianghong; Bai, Yang; Shao, Chunlin, E-mail: clshao@shmu.edu.cn

    2014-05-15

    Highlights: • Inhibition of H{sub 2}S/CSE pathway strongly stimulates cellular apoptosis. • Inhibition of H{sub 2}S/CSE pathway suppresses cell growth by blocking EGFR pathway. • H{sub 2}S/CSE pathway is critical for maintaining the proliferation of hepatoma cells. - Abstract: Hydrogen sulfide (H{sub 2}S)/cystathionine γ-lyase (CSE) pathway has been demonstrated to play vital roles in physiology and pathophysiology. However, its role in tumor cell proliferation remains largely unclear. Here we found that CSE over-expressed in hepatoma HepG2 and PLC/PRF/5 cells. Inhibition of endogenous H{sub 2}S/CSE pathway drastically decreased the proliferation of HepG2 and PLC/PRF/5 cells, and it also enhanced ROS production and mitochondrial disruption, pronounced DNA damage and increased apoptosis. Moreover, this increase of apoptosis was associated with the activation of p53 and p21 accompanied by a decreased ratio of Bcl-2/Bax and up-regulation of phosphorylated c-Jun N-terminal kinase (JNK) and caspase-3 activity. In addition, the negative regulation of cell proliferation by inhibition of H{sub 2}S/CSE system correlated with the blockage of cell mitogenic and survival signal transduction of epidermal growth factor receptor (EGFR) via down-regulating the extracellular-signal-regulated kinase 1/2 (ERK1/2) activation. These results demonstrate that H{sub 2}S/CSE and its downstream pathway contribute to the proliferation of hepatoma cells, and inhibition of this pathway strongly suppress the excessive growth of hepatoma cells by stimulating mitochondrial apoptosis and suppressing cell growth signal transduction.

  5. Biochemical characterization of a novel tyrosine phenol-lyase from Fusobacterium nucleatum for highly efficient biosynthesis of l-DOPA.

    Science.gov (United States)

    Zheng, Ren-Chao; Tang, Xiao-Ling; Suo, Hui; Feng, Li-Lin; Liu, Xiao; Yang, Jian; Zheng, Yu-Guo

    2018-05-01

    Tyrosine phenol-lyase (TPL) catalyzes the reversible cleavage of l-tyrosine to phenol, pyruvate and ammonia. When pyrocatechol is substituted for phenol, l-dihydroxyphenylalanine (l-DOPA) is produced. The TPL-catalyzed route was regarded as the most economic process for l-DOPA production. In this study, a novel TPL from Fusobacterium nucleatum (Fn-TPL) was successfully overexpressed in Escherichia coli and screened for l-DOPA synthesis with a specific activity of 2.69Umg -1 . Fn-TPL was found to be a tetramer, and the optimal temperature and pH for α, β-elimination of l-tyrosine was 60°C and pH 8.5, respectively. The enzyme showed broad substrate specificity toward natural and synthetic l-amino acids. Kinetic analysis suggested that the k cat /K m value for l-tyrosine decomposition was much higher than that for l-DOPA decomposition, while Fn-TPL exhibited similar catalytic efficiency for synthesis of l-tyrosine and l-DOPA. With whole cells of recombinant E. coli as biocatalyst, l-DOPA yield reached 110gL -1 with a pyrocatechol conversion of 95%, which was comparable to the reported highest level. The results demonstrated the great potential of Fn-TPL for industrial production of l-DOPA. Copyright © 2017 Elsevier Inc. All rights reserved.

  6. Gene deletion of cytosolic ATP: citrate lyase leads to altered organic acid production in Aspergillus niger

    DEFF Research Database (Denmark)

    Meijer, Susan Lisette; Nielsen, Michael Lynge; Olsson, Lisbeth

    2009-01-01

    With the availability of the genome sequence of the filamentous fungus Aspergillus niger, the use of targeted genetic modifications has become feasible. This, together with the fact that A. niger is well established industrially, makes this fungus an attractive micro-organism for creating a cell...... factory platform for production of chemicals. Using molecular biology techniques, this study focused on metabolic engineering of A. niger to manipulate its organic acid production in the direction of succinic acid. The gene target for complete gene deletion was cytosolic ATP: citrate lyase (acl), which...... the acl gene. Additionally, the total amount of organic acids produced in the deletion strain was significantly increased. Genome-scale stoichiometric metabolic model predictions can be used for identifying gene targets. Deletion of the acl led to increased succinic acid production by A. niger....

  7. ePAL Vision 2020 for active ageing of senior professionals

    NARCIS (Netherlands)

    Afsarmanesh, H.; Msanjila, S.S.

    2010-01-01

    In order to enhance the active life of senior professionals, one fundamental challenge is to identify ways to assist promoting the role of elder people within the continuously ageing European society. This paper proposes a vision for establishing required support environment for communities of

  8. Isolation and amino acid sequence of a dehydratase acting on d-erythro-3-hydroxyaspartate from Pseudomonas sp. N99, and its application in the production of optically active 3-hydroxyaspartate.

    Science.gov (United States)

    Nagano, Hiroyuki; Shibano, Kana; Matsumoto, Yu; Yokota, Atsushi; Wada, Masaru

    2017-06-01

    An enzyme catalyzing the ammonia-lyase reaction for the conversion of d-erythro-3-hydroxyaspartate to oxaloacetate was purified from the cell-free extract of a soil-isolated bacterium Pseudomonas sp. N99. The enzyme exhibited ammonia-lyase activity toward l-threo-3-hydroxyaspartate and d-erythro-3-hydroxyaspartate, but not toward other 3-hydroxyaspartate isomers. The deduced amino acid sequence of the enzyme, which belongs to the serine/threonine dehydratase family, shows similarity to the sequence of l-threo-3-hydroxyaspartate ammonia-lyase (EC 4.3.1.16) from Pseudomonas sp. T62 (74%) and Saccharomyces cerevisiae (64%) and serine racemase from Schizosaccharomyces pombe (65%). These results suggest that the enzyme is similar to l-threo-3-hydroxyaspartate ammonia-lyase from Pseudomonas sp. T62, which does not act on d-erythro-3-hydroxyaspartate. We also then used the recombinant enzyme expressed in Escherichia coli to produce optically pure l-erythro-3-hydroxyaspartate and d-threo-3-hydroxyaspartate from the corresponding dl-racemic mixtures. The enzymatic resolution reported here is one of the simplest and the first enzymatic method that can be used for obtaining optically pure l-erythro-3-hydroxyaspartate.

  9. A Pectate Lyase-Coding Gene Abundantly Expressed during Early Stages of Infection Is Required for Full Virulence in Alternaria brassicicola.

    Directory of Open Access Journals (Sweden)

    Yangrae Cho

    Full Text Available Alternaria brassicicola causes black spot disease of Brassica species. The functional importance of pectin digestion enzymes and unidentified phytotoxins in fungal pathogenesis has been suspected but not verified in A. brassicicola. The fungal transcription factor AbPf2 is essential for pathogenicity and induces 106 genes during early pathogenesis, including the pectate lyase-coding gene, PL1332. The aim of this study was to test the importance and roles of PL1332 in pathogenesis. We generated deletion strains of the PL1332 gene, produced heterologous PL1332 proteins, and evaluated their association with virulence. Deletion strains of the PL1332 gene were approximately 30% less virulent than wild-type A. brassicicola, without showing differences in colony expansion on solid media and mycelial growth in nutrient-rich liquid media or minimal media with pectins as a major carbon source. Heterologous PL1332 expressed as fusion proteins digested polygalacturons in vitro. When the fusion proteins were injected into the apoplast between leaf veins of host plants the tissues turned dark brown and soft, resembling necrotic leaf tissue. The PL1332 gene was the first example identified as a general toxin-coding gene and virulence factor among the 106 genes regulated by the transcription factor, AbPf2. It was also the first gene to have its functions investigated among the 19 pectate lyase genes and several hundred putative cell-wall degrading enzymes in A. brassicicola. These results further support the importance of the AbPf2 gene as a key pathogenesis regulator and possible target for agrochemical development.

  10. Paraffin as oxygen vector modulates tyrosine phenol lyase production by Citrobacter freundii MTCC 2424.

    Science.gov (United States)

    Azmi, Wamik; Kumar, Ajay; Dev, Varun

    2013-06-01

    The efficiency of three oxygen-vectors liquid paraffin, silicone oil and n-dodecane in the production of tyrosine phenol lyase (TPL) by Citrobacter freundii MTCC 2424 was evaluated at 4% (v/v) concentration. The liquid paraffin as oxygenvectors was found to exhibit a stimulatory effect on TPL synthesis. The liquid paraffin at 6% (v/v) resulted in 34% increase in the TPL synthesis accompanied by a 13% increase in the production of cell mass at a 10 L scale. This improvement in TPL and cell mass production in the presence of liquid paraffin can be related to the fact that liquid paraffin was capable of maintaining dissolved O2 concentration above 28% throughout the course of the fermentation. Maintenance of the dissolved O2 concentration above 28% could be viewed in terms of an adequate oxygen supply to the rapidly dividing cells of the bacterium, which in turn resulted in enhanced synthesis of TPL and cell mass.

  11. Structural and Biochemical Characterization of a Copper-Binding Mutant of the Organomercurial Lyase MerB: Insight into the Key Role of the Active Site Aspartic Acid in Hg-Carbon Bond Cleavage and Metal Binding Specificity.

    Science.gov (United States)

    Wahba, Haytham M; Lecoq, Lauriane; Stevenson, Michael; Mansour, Ahmed; Cappadocia, Laurent; Lafrance-Vanasse, Julien; Wilkinson, Kevin J; Sygusch, Jurgen; Wilcox, Dean E; Omichinski, James G

    2016-02-23

    In bacterial resistance to mercury, the organomercurial lyase (MerB) plays a key role in the detoxification pathway through its ability to cleave Hg-carbon bonds. Two cysteines (C96 and C159; Escherichia coli MerB numbering) and an aspartic acid (D99) have been identified as the key catalytic residues, and these three residues are conserved in all but four known MerB variants, where the aspartic acid is replaced with a serine. To understand the role of the active site serine, we characterized the structure and metal binding properties of an E. coli MerB mutant with a serine substituted for D99 (MerB D99S) as well as one of the native MerB variants containing a serine residue in the active site (Bacillus megaterium MerB2). Surprisingly, the MerB D99S protein copurified with a bound metal that was determined to be Cu(II) from UV-vis absorption, inductively coupled plasma mass spectrometry, nuclear magnetic resonance, and electron paramagnetic resonance studies. X-ray structural studies revealed that the Cu(II) is bound to the active site cysteine residues of MerB D99S, but that it is displaced following the addition of either an organomercurial substrate or an ionic mercury product. In contrast, the B. megaterium MerB2 protein does not copurify with copper, but the structure of the B. megaterium MerB2-Hg complex is highly similar to the structure of the MerB D99S-Hg complexes. These results demonstrate that the active site aspartic acid is crucial for both the enzymatic activity and metal binding specificity of MerB proteins and suggest a possible functional relationship between MerB and its only known structural homologue, the copper-binding protein NosL.

  12. Cinatii, Rambois e Pereira Câo e a pintura-decorativa do Palácio Angeje-Palmela no Paço do Lumiar

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    Miguel Leal

    2006-01-01

    Full Text Available Na zona do Paço do Lumiar, outrora fora de Lisboa, encontra-se o Palácio Angeja- Palmela, edifício palaciano do século XVIII, construído no post terramoto de 1755. Nos seus interiores encontramos uma decoração realizada em pintura a fresco e que podemos atribuir à tríade Cinatti, Rambois e Pereira Cão, cenógrafos e pintores-decoradores da Lisboa de Oitocentos. O palácio, os seus jardins e a sua decoração interior respiram uma atmosfera romântica que é visível na pintura decorativa dos seus interiores, encomenda do seu antigo proprietário D. Domingos de Sousa Holstein, segundo duque de Palmela.In the site of Paço do Lumiar, we find the Angeja-Palmela palace, a building of the XVIII century, built after the Lisbon earthquake of 1755. Inside the palace we find a fresh paintings decoration made by Cinatti, Rambois and Pereira Cão, Lisbon´s set designers and decorative painters of the XIX century. The palace, the gardens and it’s interior decoration breath a romantic atmosphere that we perceive in the respective mural paintings, a commission of it’s former owner, D. Domigos de Sousa Holstein, second duke of Palmela.

  13. Influence of Goal Setting on Physical Activity and Cardiorespiratory Endurance in Low-Income Children Enrolled in CSPAP Schools

    Science.gov (United States)

    Burns, Ryan D.; Brusseau, Timothy A.; Fu, You

    2017-01-01

    Background: Comprehensive school physical activity programming (CSPAP) has been shown to increase school day physical activity and health-related fitness. The use of goal setting may further enhance the outcomes of CSPAP. Purpose: The purpose of this study was to examine the effect of physical activity leader (PAL) goal setting on school day…

  14. Hours spent and energy expended in physical activity domains: Results from The Tomorrow Project cohort in Alberta, Canada

    Science.gov (United States)

    2011-01-01

    Background Knowledge of adult activity patterns across domains of physical activity is essential for the planning of population-based strategies that will increase overall energy expenditure and reduce the risk of obesity and related chronic diseases. We describe domain-specific hours of activity and energy expended among participants in a prospective cohort in Alberta, Canada. Methods The Past Year Total Physical Activity Questionnaire was completed by 15,591 Tomorrow Project® participants, between 2001 and 2005 detailing physical activity type, duration, frequency and intensity. Domain-specific hours of activity and activity-related energy expenditure, expressed as a percent of total energy expenditure (TEE) (Mean (SD); Median (IQR)) are reported across inactive (<1.4), low active (1.4 to 1.59), active (1.6 to 1.89) and very active (≥ 1.9) Physical Activity Level (PAL = TEE:REE) categories. Results In very active women and amongst all men except those classified as inactive, activity-related energy expenditure comprised primarily occupational activity. Amongst inactive men and women in active, low active and inactive groups, activity-related energy expenditure from household activity was comparable to, or exceeded that for occupational activity. Leisure-time activity-related energy expenditure decreased with decreasing PAL categories; however, even amongst the most active men and women it accounted for less than 10 percent of TEE. When stratified by employment status, leisure-time activity-related energy expenditure was greatest for retired men [mean (SD): 10.8 (8.5) percent of TEE], compared with those who were fully employed, employed part-time or not employed. Transportation-related activity was negligible across all categories of PAL and employment status. Conclusion For the inactive portion of this population, active non-leisure activities, specifically in the transportation and occupational domains, need to be considered for inclusion in daily routines

  15. Sphingosine kinase 1 is regulated by peroxisome proliferator-activated receptor α in response to free fatty acids and is essential for skeletal muscle interleukin-6 production and signaling in diet-induced obesity.

    Science.gov (United States)

    Ross, Jessica S; Hu, Wei; Rosen, Bess; Snider, Ashley J; Obeid, Lina M; Cowart, L Ashley

    2013-08-02

    We previously demonstrated that sphingosine kinase 1 (Sphk1) expression and activity are up-regulated by exogenous palmitate (PAL) in a skeletal muscle model system and in diet-induced obesity in mice; however, potential functions and in vivo relevance of this have not been addressed. Here, we aimed to determine the mechanism by which PAL regulates SphK1 in muscle, and to determine potential roles for its product, sphingosine-1-phosphate (S1P), in muscle biology in the context of obesity. Cloning and analysis of the mouse Sphk1 promoter revealed a peroxisome proliferator-activated receptor (PPAR) α cis-element that mediated activation of a reporter under control of the Sphk1 promoter; direct interaction of PPARα was demonstrated by chromatin immunoprecipitation. PAL treatment induced the proinflammatory cytokine interleukin (IL)-6 in a manner dependent on SphK1, and this was attenuated by inhibition of the sphingosine-1-phosphate receptor 3 (S1PR3). Diet-induced obesity in mice demonstrated that IL-6 expression in muscle, but not adipose tissue, increased in obesity, but this was attenuated in Sphk1(-/-) mice. Moreover, plasma IL-6 levels were significantly decreased in obese Sphk1(-/-) mice relative to obese wild type mice, and muscle, but not adipose tissue IL-6 signaling was activated. These data indicate that PPARα regulates Sphk1 expression in the context of fatty acid oversupply and links PAL to muscle IL-6 production. Moreover, this function of SphK1 in diet-induced obesity suggests a potential role for SphK1 in obesity-associated pathological outcomes.

  16. Physcomitrella patens activates reinforcement of the cell wall, programmed cell death and accumulation of evolutionary conserved defence signals, such as salicylic acid and 12-oxo-phytodienoic acid, but not jasmonic acid, upon Botrytis cinerea infection.

    Science.gov (United States)

    Ponce De León, Inés; Schmelz, Eric A; Gaggero, Carina; Castro, Alexandra; Álvarez, Alfonso; Montesano, Marcos

    2012-10-01

    The moss Physcomitrella patens is an evolutionarily basal model system suitable for the analysis of plant defence responses activated after pathogen assault. Upon infection with the necrotroph Botrytis cinerea, several defence mechanisms are induced in P. patens, including the fortification of the plant cell wall by the incorporation of phenolic compounds and the induced expression of related genes. Botrytis cinerea infection also activates the accumulation of reactive oxygen species and cell death with hallmarks of programmed cell death in moss tissues. Salicylic acid (SA) levels also increase after fungal infection, and treatment with SA enhances transcript accumulation of the defence gene phenylalanine ammonia-lyase (PAL) in P. patens colonies. The expression levels of the genes involved in 12-oxo-phytodienoic acid (OPDA) synthesis, including lipoxygenase (LOX) and allene oxide synthase (AOS), increase in P. patens gametophytes after pathogen assault, together with a rise in free linolenic acid and OPDA concentrations. However, jasmonic acid (JA) could not be detected in healthy or infected tissues of this plant. Our results suggest that, although conserved defence signals, such as SA and OPDA, are synthesized and are probably involved in the defence response of P. patens against B. cinerea infection, JA production appears to be missing. Interestingly, P. patens responds to OPDA and methyl jasmonate by reducing moss colony growth and rhizoid length, suggesting that jasmonate perception is present in mosses. Thus, P. patens can provide clues with regard to the evolution of different defence pathways in plants, including signalling and perception of OPDA and jasmonates in nonflowering and flowering plants. © 2012 THE AUTHORS. MOLECULAR PLANT PATHOLOGY © 2012 BSPP AND BLACKWELL PUBLISHING LTD.

  17. Production of Pectate Lyase by Penicillium viridicatum RFC3 in Solid-State and Submerged Fermentation

    Directory of Open Access Journals (Sweden)

    Viviani Ferreira

    2010-01-01

    Full Text Available Pectate lyase (PL was produced by the filamentous fungus Penicillium viridicatum RFC3 in solid-state cultures of a mixture of orange bagasse and wheat bran (1 : 1 w/w, or orange bagasse, wheat bran and sugarcane bagasse (1 : 1 : 0.5 w/w, and in a submerged liquid culture with orange bagasse and wheat bran (3% as the carbon source. PL production was highest (1,500 U  mL−1 or 300 Ug−1 of substrate in solid-state fermentation (SSF on wheat bran and orange bagasse at 96 hours. PL production in submerged fermentation (SmF was influenced by the initial pH of the medium. With the initial pH adjusted to 4.5, 5.0, and 5.5, the peak activity was observed after 72, 48, and 24 hours of fermentation, respectively, when the pH of the medium reached the value 5.0. PL from SSF and SmF were loaded on Sephadex-G75 columns and six activity peaks were obtained from crude enzyme from SSF and designated PL I, II, III, IV, V, and VI, while five peaks were obtained from crude enzyme from SmF and labeled PL  I′, II′, III′, IV′, and VII′. Crude enzyme and fraction III from each fermentative process were tested further. The optimum pH for crude PL from either process was 5.5, while that for PL III was 8.0. The maximum activity of enzymes from SSF was observed at 35∘C, but crude enzyme was more thermotolerant than PL III, maintaining its maximum activity up to 45∘C. Crude enzyme from SmF and PL   III′ showed thermophilic profiles of activity, with maximum activity at 60 and 55∘C, respectively. In the absence of substrate, the crude enzyme from SSF was stable over the pH range 3.0–10.0 and PL III was most stable in the pH range 4.0–7.0. Crude enzyme from SmF retained 70%–80% of its maximum activity in the acid-neutral pH range (4.0–7.0, but PIII showed high stability at alkaline pH (7.5–9.5. PL from SSF was more thermolabile than that from SmF. The latter maintained 60% of its initial activity after 1 h at 55

  18. Production of Pectate Lyase by Penicillium viridicatum RFC3 in Solid-State and Submerged Fermentation

    Science.gov (United States)

    Ferreira, Viviani; da Silva, Roberto; Silva, Dênis; Gomes, Eleni

    2010-01-01

    Pectate lyase (PL) was produced by the filamentous fungus Penicillium viridicatum RFC3 in solid-state cultures of a mixture of orange bagasse and wheat bran (1 : 1 w/w), or orange bagasse, wheat bran and sugarcane bagasse (1 : 1 : 0.5 w/w), and in a submerged liquid culture with orange bagasse and wheat bran (3%) as the carbon source. PL production was highest (1,500 U  mL−1 or 300 Ug−1 of substrate) in solid-state fermentation (SSF) on wheat bran and orange bagasse at 96 hours. PL production in submerged fermentation (SmF) was influenced by the initial pH of the medium. With the initial pH adjusted to 4.5, 5.0, and 5.5, the peak activity was observed after 72, 48, and 24 hours of fermentation, respectively, when the pH of the medium reached the value 5.0. PL from SSF and SmF were loaded on Sephadex-G75 columns and six activity peaks were obtained from crude enzyme from SSF and designated PL I, II, III, IV, V, and VI, while five peaks were obtained from crude enzyme from SmF and labeled PL  I′, II′, III′, IV′, and VII′. Crude enzyme and fraction III from each fermentative process were tested further. The optimum pH for crude PL from either process was 5.5, while that for PL III was 8.0. The maximum activity of enzymes from SSF was observed at 35°C, but crude enzyme was more thermotolerant than PL III, maintaining its maximum activity up to 45°C. Crude enzyme from SmF and PL III′ showed thermophilic profiles of activity, with maximum activity at 60 and 55°C, respectively. In the absence of substrate, the crude enzyme from SSF was stable over the pH range 3.0–10.0 and PL III was most stable in the pH range 4.0–7.0. Crude enzyme from SmF retained 70%–80% of its maximum activity in the acid-neutral pH range (4.0–7.0), but PIII showed high stability at alkaline pH (7.5–9.5). PL from SSF was more thermolabile than that from SmF. The latter maintained 60% of its initial activity after 1 h at 55°C. The differing

  19. A Look at the Researches of the Syro-Italian Archaeological Mission in Palmyra (PAL.M.A.I.S.

    Directory of Open Access Journals (Sweden)

    Gioia Zenoni

    2014-08-01

    Full Text Available Dal 2007 la Missione Archeologica Italo-Siriana a Palmira (PAL.M.A.I.S., nata da un accordo fra l’Università degli Studi di Milano e la Direzione Generale delle Antichità e dei Musei di Damasco, opera nel sito di Tadmor-Palmira (Siria sotto la direzione della prof. Maria Teresa Grassi. Le ricerche, finalizzate allo studio dell’edilizia residenziale privata della città romana attraverso l’indagine di un settore cruciale, il Quartiere Sud-Ovest, sono state supportate da attività sul campo - fra cui lo scavo stratigrafico dell’Edificio con Peristilio, ora interrotto - e dal 2011 si sono concentrate sullo studio dell’architettura e della cultura materiale in vista della pubblicazione finale.

  20. One Year Experience of Pheburane(®) (Sodium Phenylbutyrate) Treatment in a Patient with Argininosuccinate Lyase Deficiency.

    Science.gov (United States)

    Uçar, Sema Kalkan; Ozbaran, Burcu; Altinok, Yasemin Atik; Kose, Melis; Canda, Ebru; Kagnici, Mehtap; Coker, Mahmut

    2015-01-01

    Argininosuccinate lyase deficiency (ASLD) is a urea cycle disorder (UCD) treated with dietary adjustment and nitrogen scavenging agents. "Pheburane(®)" is a new tasteless and odour-free formulation of sodium phenylbutyrate, indicated in the treatment of UCD.A male patient diagnosed with ASLD was put on treatment with the new formulation of sodium phenylbutyrate (granules) for a period of one year, at 500 mg/kg orally in 3 intakes/day. Plasma glutamine, arginine, citrulline, argininosuccinate, serum sodium, potassium, liver function tests and urine orotate all remained unchanged over this period. There was no difference in mean ammonia levels before and after treatment, and no hyperammonemia episode occurred during treatment with Pheburane(®). An improvement in a measurement of quality of life (QOL) was noted after treatment with Pheburane(®). Good metabolic control and improved QOL were achieved throughout the treatment period.

  1. Effect of pyrogallol on the physiology and biochemistry of litchi fruit during storage

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    Jing Guoxing

    2013-01-01

    Full Text Available Abstract Background Litchi (Litchi chinensis Sonn. fruit are highly perishable and have a very short shelf life, easily turning brown and decaying. This study investigated the efficiency of pyrogallol, a catechin on the physiology and biochemistry in relation to storage life of litchi fruit. Results Fruit were treated with pyrogallol at 1 mM and then stored at ambient temperature (25°C or low temperature (4°C. Compared with control, pyrogallol significantly reduced pericarp browning and delayed the rotting of fruit day 4 at 25°C, and on day 30 at 4°C. The chemical treatment reduced respiration rate and the activities of peroxidase (POD and polyphenol oxidase (PPO, and delayed the loss of membrane permeability. Pyrogallol increased the activity of phenylalanine ammonia-lyase (PAL, delayed the loss of anthocyanin and phenolics, and maintained high 2,2-diphenyl-1-picrlhydrazyl (DPPH radical scavenging activity and reducing power. High performance liquid chromatograph (HPLC analysis clearly indicated that treated fruit contained higher concentration of the four phenolic compounds procyanidin B1, (+-catechin, (−-epicatechin and (−-epicatechin-3-gallate. Conclusions The application of pyrogallol partially reducing pericarp browning and changed quality-related physiological activities and, thus, pyrogallol could have beneficial effects on pericarp browning and fruit decay control, and could be helpful for litchi fruit postharvest storage.

  2. Évolution des politiques environnementales et tourisme de montagne au Népal

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    Isabelle Sacareau

    2009-12-01

    Full Text Available Le développement du trekking au Népal a suscité des inquiétudes qui ont abouti à la création d’aires protégées sur la foi d’un scénario de crise environnementale dont les touristes et les paysans étaient jugés responsables. Devant les conflits entre la conservation et les nécessités de développement des régions concernées, la gouvernance des aires protégées a évolué dans le sens d’une meilleure prise en compte des sociétés locales. Pour autant les mesures prises ne font qu’accompagner et diriger des dynamiques touristiques plus anciennes qui s’exercent à des échelles plus vastes. Le trekking constitue ainsi un système touristique très largement aux mains des sociétés montagnardes du pays qui a permis l’amélioration de leurs conditions de vie tout en limitant ses impacts environnementaux. En ce sens le trekking répond assez largement aux principes du développement durable.Against a backdrop of environmental crisis, attributed to the impact of tourism and the practices of rural populations, Nepal has created protected areas with a view to preserving the Himalayan environment and promoting sustainable tourism in the regions concerned. Given the conflicts between conservation needs and development needs, local communities are now being given a bigger role in the governance of these protected areas. Yet the measures being taken simply accompany and guide well-established tourism dynamics that operate on a much greater scale. Trekking is thus a tourism system largely in the hands of the country’s mountain communities and is an activity that has enabled these communities to improve their living conditions while at the same time limiting environmental impacts. In this sense it is very much in line with the principles of sustainable development.

  3. Probing Reversible Chemistry in Coenzyme B12-Dependent Ethanolamine Ammonia Lyase with Kinetic Isotope Effects

    Science.gov (United States)

    Jones, Alex R; Rentergent, Julius; Scrutton, Nigel S; Hay, Sam

    2015-01-01

    Coenzyme B12-dependent enzymes such as ethanolamine ammonia lyase have remarkable catalytic power and some unique properties that enable detailed analysis of the reaction chemistry and associated dynamics. By selectively deuterating the substrate (ethanolamine) and/or the β-carbon of the 5′-deoxyadenosyl moiety of the intrinsic coenzyme B12, it was possible to experimentally probe both the forward and reverse hydrogen atom transfers between the 5′-deoxyadenosyl radical and substrate during single-turnover stopped-flow measurements. These data are interpreted within the context of a kinetic model where the 5′-deoxyadenosyl radical intermediate may be quasi-stable and rearrangement of the substrate radical is essentially irreversible. Global fitting of these data allows estimation of the intrinsic rate constants associated with CoC homolysis and initial H-abstraction steps. In contrast to previous stopped-flow studies, the apparent kinetic isotope effects are found to be relatively small. PMID:25950663

  4. Cloning, sequence and expression of the pel gene from an Amycolata sp.

    Science.gov (United States)

    Brühlmann, F; Keen, N T

    1997-11-20

    The pel gene from an Amycolata sp. encoding a pectate lyase (EC 4.2.2.2) was isolated by activity screening a genomic DNA library in Streptomyces lividans TK24. Subsequent subcloning and sequencing of a 2.3 kb BamHI BglII fragment revealed an open reading frame of 930 nt corresponding to a protein of 29,660 Da. The overall G + C content for the coding region was 65%, with a strong G + C preference in the third (wobble) codon position (93%). A putative ribosome-binding site 5'-GGGAG-3' preceded the translational start codon by 7 base pairs. The Amycolata pectate lyase contains a signal peptide of 26 amino acids, that is cleaved after the sequence Ala-Thr-Ala. The size of the deduced protein as well as its N-terminal amino-acid sequence match the wild-type pectate lyase from the Amycolata sp. Expression of the pel gene in S. lividans TK24 resulted in high pectate lyase activity in the culture supernatant, concomitant with the appearance of a dominant protein band on a sodium dodecyl polyacrylamide gel at 30 kDa. No pectate lyase activity was detected in E. coli BL21 with the pel gene under the strong T7 promotor. The deduced amino-acid sequence showed 40% identity with PelE from Erwinia chrysanthemi and the pectate lyase from Glomerella cingulata. The Amycolata pectate lyase clearly belongs to the pectate lyase superfamily, sharing all functional amino acids and likely has a similar structural topology as Pels from Erwinia chrysanthemi and Bacillus subtilis.

  5. Metabolic reduction of phenylpropanoid compounds in primary leaves of rye (Secale cereale L.) leads to increased UV-B sensitivity of photosynthesis

    International Nuclear Information System (INIS)

    Reuber, S.; Leitsch, J.; Krause, G.H.; Weissenböck, G.

    1993-01-01

    The present study was undertaken in order to investigate the suitability of certain markers for UV plant response. In addition, we attempted to link the internal tissue distribution of specific UV-absorbing compounds to profiles of radiation gradients within intact primary rye leaves (Secale cereale L. cv. Kustro). Etiolated rye seedlings irradiated with low visible light (LL) and/or UV radiation were used to study enzyme activities of the two key enzymes, phenylalanine ammonia-lyase (PAL) and chalcone synthase (CHS), together with the tissue-specific accumulation of soluble phenylpropanoid products. Plants grown under relatively high visible light (HL) with or without supplementary UV-B radiation were used for further characterization. Apparent quantum yield and fluorescence quenching parameters were monitored to assess potential physiological changes due to UV-B exposure in HL-grown seedlings. A quartz fibreoptic microprobe was used to characterize the internal UV-B gradient of the leaf. The response of the phenylpropanoid metabolism to UV radiation was similar in primary leaves of both etiolated and HL-treated green plants. The epidermis-specific flavonoids together with CHS activity turned out to be suitable markers for assessing the effect of UV on the phenolic metabolism. The functional role of phenylpropanoid compounds was strongly implicated in protecting rye from UV-B radiation

  6. S1P Lyase Regulation of Thymic Egress and Oncogenic Inflammatory Signaling

    Science.gov (United States)

    Kumar, Ashok; Zamora-Pineda, Jesus; Degagné, Emilie

    2017-01-01

    Sphingosine-1-phosphate (S1P) is a potent lipid signaling molecule that regulates pleiotropic biological functions including cell migration, survival, angiogenesis, immune cell trafficking, inflammation, and carcinogenesis. It acts as a ligand for a family of cell surface receptors. S1P concentrations are high in blood and lymph but low in tissues, especially the thymus and lymphoid organs. S1P chemotactic gradients are essential for lymphocyte egress and other aspects of physiological cell trafficking. S1P is irreversibly degraded by S1P lyase (SPL). SPL regulates lymphocyte trafficking, inflammation and other physiological and pathological processes. For example, SPL located in thymic dendritic cells acts as a metabolic gatekeeper that controls the normal egress of mature T lymphocytes from the thymus into the circulation, whereas SPL deficiency in gut epithelial cells promotes colitis and colitis-associated carcinogenesis (CAC). Recently, we identified a complex syndrome comprised of nephrosis, adrenal insufficiency, and immunological defects caused by inherited mutations in human SGPL1, the gene encoding SPL. In the present article, we review current evidence supporting the role of SPL in thymic egress, inflammation, and cancer. Lastly, we summarize recent progress in understanding other SPL functions, its role in inherited disease, and SPL targeting for therapeutic purposes. PMID:29333002

  7. PECTATE LYASE-LIKE 9 from Brassica campestris is associated with intine formation.

    Science.gov (United States)

    Jiang, Jingjing; Yao, Lina; Yu, Youjian; Liang, Ying; Jiang, Jianxia; Ye, Nenghui; Miao, Ying; Cao, Jiashu

    2014-12-01

    Brassica campestris pectate lyase-like 9 (BcPLL9) was previously identified as a differentially expressed gene both in buds during late pollen developmental stage and in pistils during fertilization in Chinese cabbage. To characterize the gene's function, antisense-RNA lines of BcPLL9 (bcpll9) were constructed in Chinese cabbage. Self- and cross-fertilization experiments harvested half seed yields when bcpll9 lines were used as pollen donors. In vivo and in vitro pollen germination assays showed that nearly half of the pollen tubes in bcpll9 were irregular with shorter length and uneven surface. Aniline blue staining identified abnormal accumulation of a specific bright blue unknown material in the bcpll9 pollen portion. Scanning electron microscopy observation verified the abnormal outthrust material to be near the pollen germinal furrows. Transmission electron microscopy observation revealed the internal endintine layer was overdeveloped and predominantly occupied the intine. This abnormally formed intine likely induced the wavy structure and growth arrest of the pollen tube in half of the bcpll9 pollen grains, which resulted in less seed yields. Collectively, this study presented a novel PLL gene that has an important function in B. campestris intine formation. Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.

  8. S1P Lyase Regulation of Thymic Egress and Oncogenic Inflammatory Signaling

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    Ashok Kumar

    2017-01-01

    Full Text Available Sphingosine-1-phosphate (S1P is a potent lipid signaling molecule that regulates pleiotropic biological functions including cell migration, survival, angiogenesis, immune cell trafficking, inflammation, and carcinogenesis. It acts as a ligand for a family of cell surface receptors. S1P concentrations are high in blood and lymph but low in tissues, especially the thymus and lymphoid organs. S1P chemotactic gradients are essential for lymphocyte egress and other aspects of physiological cell trafficking. S1P is irreversibly degraded by S1P lyase (SPL. SPL regulates lymphocyte trafficking, inflammation and other physiological and pathological processes. For example, SPL located in thymic dendritic cells acts as a metabolic gatekeeper that controls the normal egress of mature T lymphocytes from the thymus into the circulation, whereas SPL deficiency in gut epithelial cells promotes colitis and colitis-associated carcinogenesis (CAC. Recently, we identified a complex syndrome comprised of nephrosis, adrenal insufficiency, and immunological defects caused by inherited mutations in human SGPL1, the gene encoding SPL. In the present article, we review current evidence supporting the role of SPL in thymic egress, inflammation, and cancer. Lastly, we summarize recent progress in understanding other SPL functions, its role in inherited disease, and SPL targeting for therapeutic purposes.

  9. Activity of adenylate cyclase in plasma membranes of pulmonary tissue remote times following nonlethal gamma-irradiation of rats

    International Nuclear Information System (INIS)

    Slozhenkina, L.V.; Ruda, V.P.; Ushakova, T.E.; Kuzin, A.M.

    1990-01-01

    Basal and stimulated activity of adenylate cyclase (cyclizing ATP-pyrophosphate lyase, E.C. 4.6.1.1., AC) in plasma membranes of pumonary tissye was being studied during a year after fractionated irradiation of rats (2 Gyx3). Basal and hormone-stimulated activity of AC was shown to vary significantly from normal 6 and 12 months after irradiation. The exposed membranes responded differently to AC activation by isoproterenol and F -

  10. Physical activity and modernization among Bolivian Amerindians.

    Science.gov (United States)

    Gurven, Michael; Jaeggi, Adrian V; Kaplan, Hillard; Cummings, Daniel

    2013-01-01

    Physical inactivity is a growing public health problem, and the fourth leading risk factor for global mortality. Conversely, indigenous populations living traditional lifestyles reportedly engage in vigorous daily activity that is protective against non-communicable diseases. Here we analyze physical activity patterns among the Tsimane, forager-horticulturalists of Amazonian Bolivia with minimal heart disease and diabetes. We assess age patterns of adult activity among men and women, test whether modernization affects activity levels, and examine whether nascent obesity is associated with reduced activity. A factorial method based on a large sample of behavioral observations was employed to estimate effects of age, sex, body mass index, and modernization variables on physical activity ratio (PAR), the ratio of total energy expenditure to basal metabolic rate. Accelerometry combined with heart rate monitoring was compared to the factorial method and used for nighttime sampling. Tsimane men and women display 24 hr physical activity level (PAL) of 2.02-2.15 and 1.73-1.85, respectively. Little time was spent "sedentary", whereas most activity was light to moderate, rather than vigorous. Activity peaks by the late twenties in men, and declines thereafter, but remains constant among women after the early teens. Neither BMI, fat free mass or body fat percentage are associated with PAR. There was no negative effect of modernization on physical activity. Tsimane display relatively high PALs typical of other subsistence populations, but of moderate intensity, and not outside the range of developed populations. Despite rapidly increasing socioeconomic change, there is little evidence that total activity has yet been affected. Overweight and obesity are more prevalent among women than men, and Spanish fluency is associated with greater obesity in women. The lack of cardiovascular disease among Tsimane is unlikely caused by activity alone; further study of diet, food intake and

  11. Physical activity and modernization among Bolivian Amerindians.

    Directory of Open Access Journals (Sweden)

    Michael Gurven

    Full Text Available Physical inactivity is a growing public health problem, and the fourth leading risk factor for global mortality. Conversely, indigenous populations living traditional lifestyles reportedly engage in vigorous daily activity that is protective against non-communicable diseases. Here we analyze physical activity patterns among the Tsimane, forager-horticulturalists of Amazonian Bolivia with minimal heart disease and diabetes. We assess age patterns of adult activity among men and women, test whether modernization affects activity levels, and examine whether nascent obesity is associated with reduced activity.A factorial method based on a large sample of behavioral observations was employed to estimate effects of age, sex, body mass index, and modernization variables on physical activity ratio (PAR, the ratio of total energy expenditure to basal metabolic rate. Accelerometry combined with heart rate monitoring was compared to the factorial method and used for nighttime sampling. Tsimane men and women display 24 hr physical activity level (PAL of 2.02-2.15 and 1.73-1.85, respectively. Little time was spent "sedentary", whereas most activity was light to moderate, rather than vigorous. Activity peaks by the late twenties in men, and declines thereafter, but remains constant among women after the early teens. Neither BMI, fat free mass or body fat percentage are associated with PAR. There was no negative effect of modernization on physical activity.Tsimane display relatively high PALs typical of other subsistence populations, but of moderate intensity, and not outside the range of developed populations. Despite rapidly increasing socioeconomic change, there is little evidence that total activity has yet been affected. Overweight and obesity are more prevalent among women than men, and Spanish fluency is associated with greater obesity in women. The lack of cardiovascular disease among Tsimane is unlikely caused by activity alone; further study of diet

  12. [An operational remote sensing algorithm of land surface evapotranspiration based on NOAA PAL dataset].

    Science.gov (United States)

    Hou, Ying-Yu; He, Yan-Bo; Wang, Jian-Lin; Tian, Guo-Liang

    2009-10-01

    Based on the time series 10-day composite NOAA Pathfinder AVHRR Land (PAL) dataset (8 km x 8 km), and by using land surface energy balance equation and "VI-Ts" (vegetation index-land surface temperature) method, a new algorithm of land surface evapotranspiration (ET) was constructed. This new algorithm did not need the support from meteorological observation data, and all of its parameters and variables were directly inversed or derived from remote sensing data. A widely accepted ET model of remote sensing, i. e., SEBS model, was chosen to validate the new algorithm. The validation test showed that both the ET and its seasonal variation trend estimated by SEBS model and our new algorithm accorded well, suggesting that the ET estimated from the new algorithm was reliable, being able to reflect the actual land surface ET. The new ET algorithm of remote sensing was practical and operational, which offered a new approach to study the spatiotemporal variation of ET in continental scale and global scale based on the long-term time series satellite remote sensing images.

  13. Simultaneous measurement of two enzyme activities using infrared spectroscopy: A comparative evaluation of PARAFAC, TUCKER and N-PLS modeling.

    Science.gov (United States)

    Baum, Andreas; Hansen, Per Waaben; Meyer, Anne S; Mikkelsen, Jørn Dalgaard

    2013-08-06

    Enzymes are used in many processes to release fermentable sugars for green production of biofuel, or the refinery of biomass for extraction of functional food ingredients such as pectin or prebiotic oligosaccharides. The complex biomasses may, however, require a multitude of specific enzymes which are active on specific substrates generating a multitude of products. In this paper we use the plant polymer, pectin, to present a method to quantify enzyme activity of two pectolytic enzymes by monitoring their superimposed spectral evolutions simultaneously. The data is analyzed by three chemometric multiway methods, namely PARAFAC, TUCKER3 and N-PLS, to establish simultaneous enzyme activity assays for pectin lyase and pectin methyl esterase. Correlation coefficients Rpred(2) for prediction test sets are 0.48, 0.96 and 0.96 for pectin lyase and 0.70, 0.89 and 0.89 for pectin methyl esterase, respectively. The retrieved models are compared and prediction test sets show that especially TUCKER3 performs well, even in comparison to the supervised regression method N-PLS. Copyright © 2013 Elsevier B.V. All rights reserved.

  14. Modeling and Re-Engineering of Azotobacter vinelandii Alginate Lyase to Enhance Its Catalytic Efficiency for Accelerating Biofilm Degradation.

    Science.gov (United States)

    Jang, Chul Ho; Piao, Yu Lan; Huang, Xiaoqin; Yoon, Eun Jeong; Park, So Hee; Lee, Kyoung; Zhan, Chang-Guo; Cho, Hoon

    2016-01-01

    Alginate is known to prevent elimination of Pseudomonas aeruginosa biofilms. Alginate lyase (AlgL) might therefore facilitate treatment of Pseudomonas aeruginosa-infected cystic fibrosis patients. However, the catalytic activity of wild-type AlgL is not sufficiently high. Therefore, molecular modeling and site-directed mutagenesis of AlgL might assist in enzyme engineering for therapeutic development. AlgL, isolated from Azotobacter vinelandii, catalyzes depolymerization of alginate via a β-elimination reaction. AlgL was modeled based on the crystal structure template of Sphingomonas AlgL species A1-III. Based on this computational analysis, AlgL was subjected to site-directed mutagenesis to improve its catalytic activity. The kcat/Km of the K194E mutant showed a nearly 5-fold increase against the acetylated alginate substrate, as compared to the wild-type. Double and triple mutants (K194E/K245D, K245D/K319A, K194E/K245D/E312D, and K194E/K245D/K319A) were also prepared. The most potent mutant was observed to be K194E/K245D/K319A, which has a 10-fold improved kcat value (against acetylated alginate) compared to the wild-type enzyme. The antibiofilm effect of both AlgL forms was identified in combination with piperacillin/tazobactam (PT) and the disruption effect was significantly higher in mutant AlgL combined with PT than wild-type AlgL. However, for both the wild-type and K194E/K245D/K319A mutant, the use of the AlgL enzyme alone did not show significant antibiofilm effect.

  15. Modeling and Re-Engineering of Azotobacter vinelandii Alginate Lyase to Enhance Its Catalytic Efficiency for Accelerating Biofilm Degradation.

    Directory of Open Access Journals (Sweden)

    Chul Ho Jang

    Full Text Available Alginate is known to prevent elimination of Pseudomonas aeruginosa biofilms. Alginate lyase (AlgL might therefore facilitate treatment of Pseudomonas aeruginosa-infected cystic fibrosis patients. However, the catalytic activity of wild-type AlgL is not sufficiently high. Therefore, molecular modeling and site-directed mutagenesis of AlgL might assist in enzyme engineering for therapeutic development. AlgL, isolated from Azotobacter vinelandii, catalyzes depolymerization of alginate via a β-elimination reaction. AlgL was modeled based on the crystal structure template of Sphingomonas AlgL species A1-III. Based on this computational analysis, AlgL was subjected to site-directed mutagenesis to improve its catalytic activity. The kcat/Km of the K194E mutant showed a nearly 5-fold increase against the acetylated alginate substrate, as compared to the wild-type. Double and triple mutants (K194E/K245D, K245D/K319A, K194E/K245D/E312D, and K194E/K245D/K319A were also prepared. The most potent mutant was observed to be K194E/K245D/K319A, which has a 10-fold improved kcat value (against acetylated alginate compared to the wild-type enzyme. The antibiofilm effect of both AlgL forms was identified in combination with piperacillin/tazobactam (PT and the disruption effect was significantly higher in mutant AlgL combined with PT than wild-type AlgL. However, for both the wild-type and K194E/K245D/K319A mutant, the use of the AlgL enzyme alone did not show significant antibiofilm effect.

  16. Visit of H.E. Mr. Attila Chikan, Minister of Economic Affairs, Prof. Adam Török, President, National Committee for Technological Development, Mr. Pal Koncz, Deputy Director General, National Committee for Technological Development, Hungary

    CERN Multimedia

    Laurent Guiraud

    1999-01-01

    Visit of H.E. Mr. Attila Chikan, Minister of Economic Affairs, Prof. Adam Török, President, National Committee for Technological Development, Mr. Pal Koncz, Deputy Director General, National Committee for Technological Development, Hungary

  17. Gaseous environment of plants and activity of enzymes of carbohydrate catabolism

    International Nuclear Information System (INIS)

    Ivanov, B.F.; Zemlyanukhin, A.A.; Igamberdiev, A.U.; Salam, A.M.M.

    1989-01-01

    The authors investigated the action of hypoxia and high CO 2 concentration in the atmosphere on activity of phosphofructokinase, aldolase, glucose phosphate isomerase, glucose-6-phosphate dehydrogenase, lactate dehydrogenase, alcohol dehydrogenase, and isocitrate lyase in pea seedlings (Pisum sativum L.), corn scutella (Zea mays L.), and hemp cotyledons (Cannabis sativa L.). The first 4-12h of hypoxia witnessed suppression of enzymes of the initial stages of glycolysis (glucose-6-phosphate isomerase, phosphofructokinase)and activation of enzymes of its final stages (alcohol dehydrogenase and lactate dehydrogenase) and enzymes linking glycolysis and the pentose phosphate pathway (aldolase and glucose-6-phosphate dehydrogenase). An excess of CO 2 in the environment accelerated and amplified this effect. At the end of a 24-h period of anaerobic incubation, deviations of enzyme activity from the control were leveled in both gaseous environments. An exception was observed in the case of phosphofructokinase, whose activity increased markedly at this time in plants exposed to CO 2 . Changes in activity of the enzymes were coupled with changes in their kinetic parameters (apparent K m and V max values). The activity of isocitrate lyase was suppressed in both variants of hypoxic gaseous environments, a finding that does not agree with the hypothesis as to participation of the glyoxylate cycle in the metabolic response of plants to oxygen stress. Thus, temporary inhibition of the system of glycolysis and activation of the pentose phosphate pathway constituted the initial response of the plants to O 2 stress, and CO 2 intensified this metabolic response

  18. Mechanistic deductions from kinetic isotope effects and pH studies of pyridoxal phosphate dependent carbon-carbon lyases: Erwinia herbicola and Citrobacter freundii tyrosine phenol-lyase

    International Nuclear Information System (INIS)

    Kiick, D.M.; Phillips, R.S.

    1988-01-01

    The pH dependence of the kinetic parameters and primary deuterium isotope effects have been determined for tyrosine phenol-lyase from both Erwinia herbicola and Citrobacter freundii. The primary deuterium isotope effects indicate that proton abstraction from the 2-position of the substrate is partially rate-limiting for both enzymes. The C. freundii enzyme primary deuterium isotope effects [DV = 3.5 and D(V/Ktyr) = 2.5] are pH independent, indicating that tyrosine is not sticky (i.e., does not dissociate slower than it reacts to give products). Since Vmax for both tyrosine and the alternate substrate S-methyl-L-cysteine is also pH independent, substrate binds only to the correctly protonated form of the enzyme. For the E. herbicola enzyme, both Vmax and V/K for tyrosine or S-methyl-L-cysteine are pH dependent, as well as both DV and D(V/Ktyr). Thus, while both the protonated and unprotonated enzyme can bind substrate, and may be interconverted directly, only the unprotonated Michaelis complex is catalytically competent. At pH 9.5, DV = 2.5 and D(V/Ktyr) = 1.5. However, at pH 6.4 the isotope effect on both parameters is equal to 4.1. From these data, the forward commitment factor (cf = 5.2) and catalytic ratio (cvf = 1.1) for tyrosine and S-methyl-L-cysteine (cf = 2.2, cvf = 24) are calculated. Also, the Michaelis complex partition ratio (cf/cvf) for substrate and products is calculated to be 4.7 for tyrosine and 0.1 for S-methyl-L-cysteine

  19. DL-β-aminobutyric acid-induced resistance in soybean against Aphis glycines Matsumura (Hemiptera: Aphididae.

    Directory of Open Access Journals (Sweden)

    Yunpeng Zhong

    Full Text Available Priming can improve plant innate capability to deal with the stresses caused by both biotic and abiotic factors. In this study, the effect of DL-β-amino-n-butyric acid (BABA against Aphis glycines Matsumura, the soybean aphid (SA was evaluated. We found that 25 mM BABA as a root drench had minimal adverse impact on plant growth and also efficiently protected soybean from SA infestation. In both choice and non-choice tests, SA number was significantly decreased to a low level in soybean seedlings drenched with 25 mM BABA compared to the control counterparts. BABA treatment resulted in a significant increase in the activities of several defense enzymes, such as phenylalanine ammonia-lyase (PAL, peroxidase (POX, polyphenol oxidase (PPO, chitinase (CHI, and β-1, 3-glucanase (GLU in soybean seedlings attacked by aphid. Meanwhile, the induction of 15 defense-related genes by aphid, such as AOS, CHS, MMP2, NPR1-1, NPR1-2, and PR genes, were significantly augmented in BABA-treated soybean seedlings. Our study suggest that BABA application is a promising way to enhance soybean resistance against SA.

  20. Phyllosticta musarum Infection-Induced Defences Suppress Anthracnose Disease Caused by Colletotrichum musae in Banana Fruits cv 'Embul'.

    Science.gov (United States)

    Abayasekara, C L; Adikaram, N K B; Wanigasekara, U W N P; Bandara, B M R

    2013-03-01

    Anthracnose development by Colletotrichum musae was observed to be significantly less in the fruits of the banana cultivar 'Embul' (Mysore, AAB) infected with Phyllosticta musarum than in fruits without such infections. Anthracnose disease originates from quiescent C. musae infections in the immature fruit. P. musarum incites minute, scattered spots, referred to as freckles, in the superficial tissues of immature banana peel which do not expand during maturation or ripening. P. musarum does not appear to have a direct suppressive effect on C. musae as conidia of C. musae germinate on both freckled and non-freckled fruit forming quiescent infections. Our investigations have shown that P. musarum infection induced several defence responses in fruit including the accumulation of five phytoalexins, upregulation of chitinase and β-1,3-glucanase, phenylalanine ammonia lyase (PAL) activity and cell wall lignification. (1)H and (13)C NMR spectral data of one purified phytoalexin compared closely with 4'-hydroxyanigorufone. Some of the P. musarum-induced defences that retained during ripening, restrict C. musae development at the ripe stage. This paper examines the potential of P. musarum-induced defences, in the control of anthracnose, the most destructive postharvest disease in banana.