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Sample records for terpene synthase tps

  1. Emission and Accumulation of Monoterpene and the Key Terpene Synthase (TPS) Associated with Monoterpene Biosynthesis in Osmanthus fragrans Lour

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    Zeng, Xiangling; Liu, Cai; Zheng, Riru; Cai, Xuan; Luo, Jing; Zou, Jingjing; Wang, Caiyun

    2016-01-01

    Osmanthus fragrans is an ornamental and economically important plant known for its magnificent aroma, and the most important aroma-active compounds in flowers are monoterpenes, mainly β-ocimene, linalool and linalool derivatives. To understand the molecular mechanism of monoterpene production, we analyzed the emission and accumulation patterns of these compounds and the transcript levels of the genes involved in their biosynthesis in two O. fragrans cultivars during flowering stages. The results showed that both emission and accumulation of monoterpenes varied with flower development and glycosylation had an important impact on floral linalool emission during this process. Gene expression demonstrated that the transcript levels of terpene synthase (TPS) genes probably played a key role in monoterpene production, compared to the genes in the MEP pathway. Phylogenetic analysis showed that OfTPS1 and OfTPS2 belonged to a TPS-g subfamily, and OfTPS3 and OfTPS4 clustered into a TPS-b subfamily. Their transient and stable expression in tobacco leaves suggested that OfTPS1 and OfTPS2 exclusively produced β-linalool, and trans-β-ocimene was the sole product from OfTPS3, while OfTPS4, a predictive sesquiterpene synthase, produced α-farnesene. These results indicate that OfTPS1, OfTPS2, and OfTPS3 could account for the major floral monoterpenes, linalool and trans-β-ocimene, produced in O. fragrans flowers. PMID:26793212

  2. Emission and accumulation of monoterpene and the key terpene synthase (TPS associated with monoterpene biosynthesis in Osmanthus fragrans Lour.

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    Xaingling eZeng

    2016-01-01

    Full Text Available Osmanthus fragrans is an ornamental and economically important plant known for its magnificent aroma, and the most important aroma-active compounds in flowers are monoterpenes, mainly β-ocimene, linalool and linalool derivatives. To understand the molecular mechanism of monoterpene production, we analyzed the emission and accumulation patterns of these compounds and the transcript levels of the genes involved in their biosynthesis in two O. fragrans cultivars during flowering stages. The results showed that both emission and accumulation of monoterpenes varied with flower development and glycosylation had an important impact on floral linalool emission during this process. Gene expression demonstrated that the transcript levels of terpene synthase (TPS genes probably played a key role in monoterpene production, compared to the genes in the MEP pathway. Phylogenetic analysis showed that OfTPS1 and OfTPS2 belonged to a TPS-g subfamily, and OfTPS3 and OfTPS4 clustered into a TPS-b subfamily. Their transient and stable expression in tobacco leaves suggested that OfTPS1 and OfTPS2 exclusively produced β-linalool, and trans-β-ocimene was the sole product from OfTPS3, while OfTPS4, a predictive sesquiterpene synthase, produced α-farnesene. These results indicate that OfTPS1, OfTPS2 and OfTPS3 could account for the major floral monoterpenes, linalool and trans-β-ocimene, produced in O. fragrans flowers.

  3. Terpene synthases from Cannabis sativa.

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    Judith K Booth

    Full Text Available Cannabis (Cannabis sativa plants produce and accumulate a terpene-rich resin in glandular trichomes, which are abundant on the surface of the female inflorescence. Bouquets of different monoterpenes and sesquiterpenes are important components of cannabis resin as they define some of the unique organoleptic properties and may also influence medicinal qualities of different cannabis strains and varieties. Transcriptome analysis of trichomes of the cannabis hemp variety 'Finola' revealed sequences of all stages of terpene biosynthesis. Nine cannabis terpene synthases (CsTPS were identified in subfamilies TPS-a and TPS-b. Functional characterization identified mono- and sesqui-TPS, whose products collectively comprise most of the terpenes of 'Finola' resin, including major compounds such as β-myrcene, (E-β-ocimene, (--limonene, (+-α-pinene, β-caryophyllene, and α-humulene. Transcripts associated with terpene biosynthesis are highly expressed in trichomes compared to non-resin producing tissues. Knowledge of the CsTPS gene family may offer opportunities for selection and improvement of terpene profiles of interest in different cannabis strains and varieties.

  4. Terpene synthases from Cannabis sativa.

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    Booth, Judith K; Page, Jonathan E; Bohlmann, Jörg

    2017-01-01

    Cannabis (Cannabis sativa) plants produce and accumulate a terpene-rich resin in glandular trichomes, which are abundant on the surface of the female inflorescence. Bouquets of different monoterpenes and sesquiterpenes are important components of cannabis resin as they define some of the unique organoleptic properties and may also influence medicinal qualities of different cannabis strains and varieties. Transcriptome analysis of trichomes of the cannabis hemp variety 'Finola' revealed sequences of all stages of terpene biosynthesis. Nine cannabis terpene synthases (CsTPS) were identified in subfamilies TPS-a and TPS-b. Functional characterization identified mono- and sesqui-TPS, whose products collectively comprise most of the terpenes of 'Finola' resin, including major compounds such as β-myrcene, (E)-β-ocimene, (-)-limonene, (+)-α-pinene, β-caryophyllene, and α-humulene. Transcripts associated with terpene biosynthesis are highly expressed in trichomes compared to non-resin producing tissues. Knowledge of the CsTPS gene family may offer opportunities for selection and improvement of terpene profiles of interest in different cannabis strains and varieties.

  5. Rice terpene synthase 24 (OsTPS24) encodes a jasmonate-responsive monoterpene synthase that produces an antibacterial γ-terpinene against rice pathogen.

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    Yoshitomi, Kayo; Taniguchi, Shiduku; Tanaka, Keiichiro; Uji, Yuya; Akimitsu, Kazuya; Gomi, Kenji

    2016-02-01

    Rice is one of the most important crops worldwide and is widely used as a model plant for molecular studies of monocotyledonous species. The plant hormone jasmonic acid (JA) is involved in rice-pathogen interactions. In addition, volatile compounds, including terpenes, whose production is induced by JA, are known to be involved in the rice defense system. In this study, we analyzed the JA-induced terpene synthase OsTPS24 in rice. We found that OsTPS24 was localized in chloroplasts and produced a monoterpene, γ-terpinene. The amount of γ-terpinene increased after JA treatment. γ-Terpinene had significant antibacterial activity against Xanthomonas oryzae pv. oryzae (Xoo); however, it did not show significant antifungal activity against Magnaporthe oryzae. The antibacterial activity of the γ-terpinene against Xoo was caused by damage to bacterial cell membranes. These results suggest that γ-terpinene plays an important role in JA-induced resistance against Xoo, and that it functions as an antibacterial compound in rice. Copyright © 2015 Elsevier GmbH. All rights reserved.

  6. The Eucalyptus terpene synthase gene family.

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    Külheim, Carsten; Padovan, Amanda; Hefer, Charles; Krause, Sandra T; Köllner, Tobias G; Myburg, Alexander A; Degenhardt, Jörg; Foley, William J

    2015-06-11

    Terpenoids are abundant in the foliage of Eucalyptus, providing the characteristic smell as well as being valuable economically and influencing ecological interactions. Quantitative and qualitative inter- and intra- specific variation of terpenes is common in eucalypts. The genome sequences of Eucalyptus grandis and E. globulus were mined for terpene synthase genes (TPS) and compared to other plant species. We investigated the relative expression of TPS in seven plant tissues and functionally characterized five TPS genes from E. grandis. Compared to other sequenced plant genomes, Eucalyptus grandis has the largest number of putative functional TPS genes of any sequenced plant. We discovered 113 and 106 putative functional TPS genes in E. grandis and E. globulus, respectively. All but one TPS from E. grandis were expressed in at least one of seven plant tissues examined. Genomic clusters of up to 20 genes were identified. Many TPS are expressed in tissues other than leaves which invites a re-evaluation of the function of terpenes in Eucalyptus. Our data indicate that terpenes in Eucalyptus may play a wider role in biotic and abiotic interactions than previously thought. Tissue specific expression is common and the possibility of stress induction needs further investigation. Phylogenetic comparison of the two investigated Eucalyptus species gives insight about recent evolution of different clades within the TPS gene family. While the majority of TPS genes occur in orthologous pairs some clades show evidence of recent gene duplication, as well as loss of function.

  7. The rice terpene synthase gene OsTPS19 functions as an (S)-limonene synthase in planta, and its overexpression leads to enhanced resistance to the blast fungus Magnaporthe oryzae.

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    Chen, Xujun; Chen, Hao; Yuan, Joshua S; Köllner, Tobias G; Chen, Yuying; Guo, Yufen; Zhuang, Xiaofeng; Chen, Xinlu; Zhang, Yong-Jun; Fu, Jianyu; Nebenführ, Andreas; Guo, Zejian; Chen, Feng

    2018-03-06

    Rice blast disease, caused by the fungus Magnaporthe oryzae, is the most devastating disease of rice. In our ongoing characterization of the defence mechanisms of rice plants against M. oryzae, a terpene synthase gene OsTPS19 was identified as a candidate defence gene. Here, we report the functional characterization of OsTPS19, which is up-regulated by M. oryzae infection. Overexpression of OsTPS19 in rice plants enhanced resistance against M. oryzae, while OsTPS19 RNAi lines were more susceptible to the pathogen. Metabolic analysis revealed that the production of a monoterpene (S)-limonene was increased and decreased in OsTPS19 overexpression and RNAi lines, respectively, suggesting that OsTPS19 functions as a limonene synthase in planta. This notion was further supported by in vitro enzyme assays with recombinant OsTPS19, in which OsTPS19 had both sesquiterpene activity and monoterpene synthase activity, with limonene as a major product. Furthermore, in a subcellular localization experiment, OsTPS19 was localized in plastids. OsTPS19 has a highly homologous paralog, OsTPS20, which likely resulted from a recent gene duplication event. We found that the variation in OsTPS19 and OsTPS20 enzyme activities was determined by a single amino acid in the active site cavity. The expression of OsTPS20 was not affected by M. oryzae infection. This indicates functional divergence of OsTPS19 and OsTPS20. Lastly, (S)-limonene inhibited the germination of M. oryzae spores in vitro. OsTPS19 was determined to function as an (S)-limonene synthase in rice and plays a role in defence against M. oryzae, at least partly, by inhibiting spore germination. © 2018 The Authors. Plant Biotechnology Journal published by Society for Experimental Biology and The Association of Applied Biologists and John Wiley & Sons Ltd.

  8. The Tomato Terpene Synthase Gene Family1[W][OA

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    Falara, Vasiliki; Akhtar, Tariq A.; Nguyen, Thuong T.H.; Spyropoulou, Eleni A.; Bleeker, Petra M.; Schauvinhold, Ines; Matsuba, Yuki; Bonini, Megan E.; Schilmiller, Anthony L.; Last, Robert L.; Schuurink, Robert C.; Pichersky, Eran

    2011-01-01

    Compounds of the terpenoid class play numerous roles in the interactions of plants with their environment, such as attracting pollinators and defending the plant against pests. We show here that the genome of cultivated tomato (Solanum lycopersicum) contains 44 terpene synthase (TPS) genes, including 29 that are functional or potentially functional. Of these 29 TPS genes, 26 were expressed in at least some organs or tissues of the plant. The enzymatic functions of eight of the TPS proteins were previously reported, and here we report the specific in vitro catalytic activity of 10 additional tomato terpene synthases. Many of the tomato TPS genes are found in clusters, notably on chromosomes 1, 2, 6, 8, and 10. All TPS family clades previously identified in angiosperms are also present in tomato. The largest clade of functional TPS genes found in tomato, with 12 members, is the TPS-a clade, and it appears to encode only sesquiterpene synthases, one of which is localized to the mitochondria, while the rest are likely cytosolic. A few additional sesquiterpene synthases are encoded by TPS-b clade genes. Some of the tomato sesquiterpene synthases use z,z-farnesyl diphosphate in vitro as well, or more efficiently than, the e,e-farnesyl diphosphate substrate. Genes encoding monoterpene synthases are also prevalent, and they fall into three clades: TPS-b, TPS-g, and TPS-e/f. With the exception of two enzymes involved in the synthesis of ent-kaurene, the precursor of gibberellins, no other tomato TPS genes could be demonstrated to encode diterpene synthases so far. PMID:21813655

  9. Isolation and characterization of terpene synthases in cotton (Gossypium hirsutum).

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    Yang, Chang-Qing; Wu, Xiu-Ming; Ruan, Ju-Xin; Hu, Wen-Li; Mao, Yin-Bo; Chen, Xiao-Ya; Wang, Ling-Jian

    2013-12-01

    Cotton plants accumulate gossypol and related sesquiterpene aldehydes, which function as phytoalexins against pathogens and feeding deterrents to herbivorous insects. However, to date little is known about the biosynthesis of volatile terpenes in this crop. Herein is reported that 5 monoterpenes and 11 sesquiterpenes from extracts of a glanded cotton cultivar, Gossypium hirsutum cv. CCRI12, were detected by gas chromatography-mass spectrometry (GC-MS). By EST data mining combined with Rapid Amplification of cDNA Ends (RACE), full-length cDNAs of three terpene synthases (TPSs), GhTPS1, GhTPS2 and GhTPS3 were isolated. By in vitro assays of the recombinant proteins, it was found that GhTPS1 and GhTPS2 are sesquiterpene synthases: the former converted farnesyl pyrophosphate (FPP) into β-caryophyllene and α-humulene in a ratio of 2:1, whereas the latter produced several sesquiterpenes with guaia-1(10),11-diene as the major product. By contrast, GhTPS3 is a monoterpene synthase, which produced α-pinene, β-pinene, β-phellandrene and trace amounts of other monoterpenes from geranyl pyrophosphate (GPP). The TPS activities were also supported by Virus Induced Gene Silencing (VIGS) in the cotton plant. GhTPS1 and GhTPS3 were highly expressed in the cotton plant overall, whereas GhTPS2 was expressed only in leaves. When stimulated by mechanical wounding, Verticillium dahliae (Vde) elicitor or methyl jasmonate (MeJA), production of terpenes and expression of the corresponding synthase genes were induced. These data demonstrate that the three genes account for the biosynthesis of volatile terpenes of cotton, at least of this Upland cotton. Copyright © 2013 Elsevier Ltd. All rights reserved.

  10. Genomic Analysis of Terpene Synthase Family and Functional Characterization of Seven Sesquiterpene Synthases from Citrus sinensis

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    Berta Alquézar

    2017-08-01

    Full Text Available Citrus aroma and flavor, chief traits of fruit quality, are derived from their high content in essential oils of most plant tissues, including leaves, stems, flowers, and fruits. Accumulated in secretory cavities, most components of these oils are volatile terpenes. They contribute to defense against herbivores and pathogens, and perhaps also protect tissues against abiotic stress. In spite of their importance, our understanding of the physiological, biochemical, and genetic regulation of citrus terpene volatiles is still limited. The availability of the sweet orange (Citrus sinensis L. Osbeck genome sequence allowed us to characterize for the first time the terpene synthase (TPS family in a citrus type. CsTPS is one of the largest angiosperm TPS families characterized so far, formed by 95 loci from which just 55 encode for putative functional TPSs. All TPS angiosperm families, TPS-a, TPS-b, TPS-c, TPS-e/f, and TPS-g were represented in the sweet orange genome, with 28, 18, 2, 2, and 5 putative full length genes each. Additionally, sweet orange β-farnesene synthase, (Z-β-cubebene/α-copaene synthase, two β-caryophyllene synthases, and three multiproduct enzymes yielding β-cadinene/α-copaene, β-elemene, and β-cadinene/ledene/allo-aromandendrene as major products were identified, and functionally characterized via in vivo recombinant Escherichia coli assays.

  11. Functional Genomics Reveals That a Compact Terpene Synthase Gene Family Can Account for Terpene Volatile Production in Apple1[W

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    Nieuwenhuizen, Niels J.; Green, Sol A.; Chen, Xiuyin; Bailleul, Estelle J.D.; Matich, Adam J.; Wang, Mindy Y.; Atkinson, Ross G.

    2013-01-01

    Terpenes are specialized plant metabolites that act as attractants to pollinators and as defensive compounds against pathogens and herbivores, but they also play an important role in determining the quality of horticultural food products. We show that the genome of cultivated apple (Malus domestica) contains 55 putative terpene synthase (TPS) genes, of which only 10 are predicted to be functional. This low number of predicted functional TPS genes compared with other plant species was supported by the identification of only eight potentially functional TPS enzymes in apple ‘Royal Gala’ expressed sequence tag databases, including the previously characterized apple (E,E)-α-farnesene synthase. In planta functional characterization of these TPS enzymes showed that they could account for the majority of terpene volatiles produced in cv Royal Gala, including the sesquiterpenes germacrene-D and (E)-β-caryophyllene, the monoterpenes linalool and α-pinene, and the homoterpene (E)-4,8-dimethyl-1,3,7-nonatriene. Relative expression analysis of the TPS genes indicated that floral and vegetative tissues were the primary sites of terpene production in cv Royal Gala. However, production of cv Royal Gala floral-specific terpenes and TPS genes was observed in the fruit of some heritage apple cultivars. Our results suggest that the apple TPS gene family has been shaped by a combination of ancestral and more recent genome-wide duplication events. The relatively small number of functional enzymes suggests that the remaining terpenes produced in floral and vegetative and fruit tissues are maintained under a positive selective pressure, while the small number of terpenes found in the fruit of modern cultivars may be related to commercial breeding strategies. PMID:23256150

  12. Identification, Functional Characterization, and Evolution of Terpene Synthases from a Basal Dicot1[OPEN

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    Yahyaa, Mosaab; Matsuba, Yuki; Brandt, Wolfgang; Doron-Faigenboim, Adi; Bar, Einat; McClain, Alan; Davidovich-Rikanati, Rachel; Lewinsohn, Efraim; Pichersky, Eran; Ibdah, Mwafaq

    2015-01-01

    Bay laurel (Laurus nobilis) is an agriculturally and economically important dioecious tree in the basal dicot family Lauraceae used in food and drugs and in the cosmetics industry. Bay leaves, with their abundant monoterpenes and sesquiterpenes, are used to impart flavor and aroma to food, and have also drawn attention in recent years because of their potential pharmaceutical applications. To identify terpene synthases (TPSs) involved in the production of these volatile terpenes, we performed RNA sequencing to profile the transcriptome of L. nobilis leaves. Bioinformatic analysis led to the identification of eight TPS complementary DNAs. We characterized the enzymes encoded by three of these complementary DNAs: a monoterpene synthase that belongs to the TPS-b clade catalyzes the formation of mostly 1,8-cineole; a sesquiterpene synthase belonging to the TPS-a clade catalyzes the formation of mainly cadinenes; and a diterpene synthase of the TPS-e/f clade catalyzes the formation of geranyllinalool. Comparison of the sequences of these three TPSs indicated that the TPS-a and TPS-b clades of the TPS gene family evolved early in the evolution of the angiosperm lineage, and that geranyllinalool synthase activity is the likely ancestral function in angiosperms of genes belonging to an ancient TPS-e/f subclade that diverged from the kaurene synthase gene lineages before the split of angiosperms and gymnosperms. PMID:26157114

  13. Genome-wide identification, functional and evolutionary analysis of terpene synthases in pineapple.

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    Chen, Xiaoe; Yang, Wei; Zhang, Liqin; Wu, Xianmiao; Cheng, Tian; Li, Guanglin

    2017-10-01

    Terpene synthases (TPSs) are vital for the biosynthesis of active terpenoids, which have important physiological, ecological and medicinal value. Although terpenoids have been reported in pineapple (Ananas comosus), genome-wide investigations of the TPS genes responsible for pineapple terpenoid synthesis are still lacking. By integrating pineapple genome and proteome data, twenty-one putative terpene synthase genes were found in pineapple and divided into five subfamilies. Tandem duplication is the cause of TPS gene family duplication. Furthermore, functional differentiation between each TPS subfamily may have occurred for several reasons. Sixty-two key amino acid sites were identified as being type-II functionally divergence between TPS-a and TPS-c subfamily. Finally, coevolution analysis indicated that multiple amino acid residues are involved in coevolutionary processes. In addition, the enzyme activity of two TPSs were tested. This genome-wide identification, functional and evolutionary analysis of pineapple TPS genes provide a new insight into understanding the roles of TPS family and lay the basis for further characterizing the function and evolution of TPS gene family. Copyright © 2017 Elsevier Ltd. All rights reserved.

  14. Rapid Discovery and Functional Characterization of Terpene Synthases from Four Endophytic Xylariaceae.

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    Weihua Wu

    Full Text Available Endophytic fungi are ubiquitous plant endosymbionts that establish complex and poorly understood relationships with their host organisms. Many endophytic fungi are known to produce a wide spectrum of volatile organic compounds (VOCs with potential energy applications, which have been described as "mycodiesel". Many of these mycodiesel hydrocarbons are terpenes, a chemically diverse class of compounds produced by many plants, fungi, and bacteria. Due to their high energy densities, terpenes, such as pinene and bisabolene, are actively being investigated as potential "drop-in" biofuels for replacing diesel and aviation fuel. In this study, we rapidly discovered and characterized 26 terpene synthases (TPSs derived from four endophytic fungi known to produce mycodiesel hydrocarbons. The TPS genes were expressed in an E. coli strain harboring a heterologous mevalonate pathway designed to enhance terpene production, and their product profiles were determined using Solid Phase Micro-Extraction (SPME and GC-MS. Out of the 26 TPS's profiled, 12 TPS's were functional, with the majority of them exhibiting both monoterpene and sesquiterpene synthase activity.

  15. The variability of sesquiterpenes emitted from two Zea mays cultivars is controlled by allelic variation of two terpene synthase genes encoding stereoselective multiple product enzymes.

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    Köllner, Tobias G; Schnee, Christiane; Gershenzon, Jonathan; Degenhardt, Jörg

    2004-05-01

    The mature leaves and husks of Zea mays release a complex blend of terpene volatiles after anthesis consisting predominantly of bisabolane-, sesquithujane-, and bergamotane-type sesquiterpenes. The varieties B73 and Delprim release the same volatile constituents but in significantly different proportions. To study the molecular genetic and biochemical mechanisms controlling terpene diversity and distribution in these varieties, we isolated the closely related terpene synthase genes terpene synthase4 (tps4) and tps5 from both varieties. The encoded enzymes, TPS4 and TPS5, each formed the same complex mixture of sesquiterpenes from the precursor farnesyl diphosphate but with different proportions of products. These mixtures correspond to the sesquiterpene blends observed in the varieties B73 and Delprim, respectively. The differences in the stereoselectivity of TPS4 and TPS5 are determined by four amino acid substitutions with the most important being a Gly instead of an Ala residue at position 409 at the catalytic site of the enzyme. Although both varieties contain tps4 and tps5 alleles, their differences in terpene composition result from the fact that B73 has only a single functional allele of tps4 and no functional alleles of tps5, whereas Delprim has only a functional allele of tps5 and no functional alleles of tps4. Lack of functionality was shown to be attributable to frame-shift mutations or amino acid substitutions that greatly reduce the activity of their encoded proteins. Therefore, the diversity of sesquiterpenes in these two maize cultivars is strongly influenced by single nucleotide changes in the alleles of two terpene synthase genes.

  16. RNA sequencing on Amomum villosum Lour. induced by MeJA identifies the genes of WRKY and terpene synthases involved in terpene biosynthesis.

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    He, Xueying; Wang, Huan; Yang, Jinfen; Deng, Ke; Wang, Teng

    2018-02-01

    Amomum villosum Lour. is an important Chinese medicinal plant that has diverse medicinal functions, and mainly contains volatile terpenes. This study aims to explore the WRKY transcription factors (TFs) and terpene synthase (TPS) unigenes that might be involved in terpene biosynthesis in A. villosum, and thus providing some new information on the regulation of terpenes in plants. RNA sequencing of A. villosum induced by methyl jasmonate (MeJA) revealed that the WRKY family was the second largest TF family in the transcriptome. Thirty-six complete WRKY domain sequences were expressed in response to MeJA. Further, six WRKY unigenes were highly correlated with eight deduced TPS unigenes. Ultimately, we combined the terpene abundance with the expression of candidate WRKY TFs and TPS unigenes to presume a possible model wherein AvWRKY61, AvWRKY28, and AvWRKY40 might coordinately trans-activate the AvNeoD promoter. We propose an approach to further investigate TF unigenes that might be involved in terpenoid biosynthesis, and identified four unigenes for further analyses.

  17. Multi-substrate terpene synthases: their occurrence and physiological significance

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    Leila Pazouki

    2016-07-01

    Full Text Available Terpene synthases are responsible for synthesis of a large number of terpenes in plants using substrates provided by two distinct metabolic pathways, the mevalonate-dependent pathway that is located in cytosol and has been suggested to be responsible for synthesis of sesquiterpenes (C15, and 2-C-methyl-D-erythritol-4-phosphate pathway located in plastids and suggested to be responsible for the synthesis of hemi- (C5, mono- (C10 and diterpenes (C20. Recent advances in characterization of genes and enzymes responsible for substrate and end product biosynthesis as well as efforts in metabolic engineering have demonstrated existence of a number of multi-substrate terpene synthases. This review summarizes the progress in the characterization of such multi-substrate terpene synthases and suggests that the presence of multi-substrate use might have been significantly underestimated. Multi-substrate use could lead to important changes in terpene product profiles upon substrate profile changes under perturbation of metabolism in stressed plants as well as under certain developmental stages. We therefore argue that multi-substrate use can be significant under physiological conditions and can result in complicate modifications in terpene profiles.

  18. Molecular cloning and functional characterization of three terpene synthases from unripe fruit of black pepper (Piper nigrum).

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    Jin, Zhehao; Kwon, Moonhyuk; Lee, Ah-Reum; Ro, Dae-Kyun; Wungsintaweekul, Juraithip; Kim, Soo-Un

    2018-01-15

    To identify terpene synthases (TPS) responsible for the biosynthesis of the sesquiterpenes that contribute to the characteristic flavors of black pepper (Piper nigrum), unripe peppercorn was subjected to the Illumina transcriptome sequencing. The BLAST analysis using amorpha-4,11-diene synthase as a query identified 19 sesquiterpene synthases (sesqui-TPSs), of which three full-length cDNAs (PnTPS1 through 3) were cloned. These sesqui-TPS cDNAs were expressed in E. coli to produce recombinant enzymes for in vitro assays, and also expressed in the engineered yeast strain to assess their catalytic activities in vivo. PnTPS1 produced β-caryophyllene as a main product and humulene as a minor compound, and thus was named caryophyllene synthase (PnCPS). Likewise, PnTPS2 and PnTPS3 were, respectively, named cadinol/cadinene synthase (PnCO/CDS) and germacrene D synthase (PnGDS). PnGDS expression in yeast yielded β-cadinene and α-copaene, the rearrangement products of germacrene D. Their k cat /K m values (20-37.7 s -1  mM -1 ) were comparable to those of other sesqui-TPSs. Among three PnTPSs, the transcript level of PnCPS was the highest, correlating with the predominant β-caryophyllene biosynthesis in the peppercorn. The products and rearranged products of three PnTPSs could account for about a half of the sesquiterpenes in number found in unripe peppercorn. Copyright © 2017 Elsevier Inc. All rights reserved.

  19. Differential accumulation of volatile terpene and terpene synthase mRNAs during lavender (Lavandula angustifolia and L. x intermedia) inflorescence development.

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    Guitton, Yann; Nicolè, Florence; Moja, Sandrine; Valot, Nadine; Legrand, Sylvain; Jullien, Frédéric; Legendre, Laurent

    2010-02-01

    Despite the commercial importance of Lavandula angustifolia Mill. and L. x intermedia Emeric ex Loisel floral essential oils (EOs), no information is currently available on potential changes in individual volatile organic compound (VOC) content during inflorescence development. Calyces were found to be the main sites of VOC accumulation. The 20 most abundant VOCs could be separated into three sub-groups according to their patterns of change in concentration The three groups of VOCs sequentially dominated the global scent bouquet of inflorescences, the transition between the first and second groups occurring around the opening of the first flower of the inflorescence and the one between the second and third groups at the start of seed set. Changes in calyx VOC accumulation were linked to the developmental stage of individual flowers. Leaves accumulated a smaller number of VOCs which were a subset of those seen in preflowering inflorescences. Their nature and content remained constant during the growing season. Quantitative real time polymerase chain reaction assessments of the expression of two terpene synthase (TPS) genes, LaLIMS and LaLINS, revealed similar trends between their patterns of expression and those of their VOC products. Molecular and chemical analyses suggest that changes in TPS expression occur during lavender inflorescence development and lead to changes in EO composition. Both molecular data and terpene analysis support the findings that changes in biosynthesis of terpene occurred during inflorescence development.

  20. Monoterpene and sesquiterpene synthases and the origin of terpene skeletal diversity in plants.

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    Degenhardt, Jörg; Köllner, Tobias G; Gershenzon, Jonathan

    2009-01-01

    The multitude of terpene carbon skeletons in plants is formed by enzymes known as terpene synthases. This review covers the monoterpene and sesquiterpene synthases presenting an up-to-date list of enzymes reported and evidence for their ability to form multiple products. The reaction mechanisms of these enzyme classes are described, and information on how terpene synthase proteins mediate catalysis is summarized. Correlations between specific amino acid motifs and terpene synthase function are described, including an analysis of the relationships between active site sequence and cyclization type and a discussion of whether specific protein features might facilitate multiple product formation.

  1. Transcriptome mining, functional characterization, and phylogeny of a large terpene synthase gene family in spruce (Picea spp.

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    Dullat Harpreet K

    2011-03-01

    Full Text Available Abstract Background In conifers, terpene synthases (TPSs of the gymnosperm-specific TPS-d subfamily form a diverse array of mono-, sesqui-, and diterpenoid compounds, which are components of the oleoresin secretions and volatile emissions. These compounds contribute to defence against herbivores and pathogens and perhaps also protect against abiotic stress. Results The availability of extensive transcriptome resources in the form of expressed sequence tags (ESTs and full-length cDNAs in several spruce (Picea species allowed us to estimate that a conifer genome contains at least 69 unique and transcriptionally active TPS genes. This number is comparable to the number of TPSs found in any of the sequenced and well-annotated angiosperm genomes. We functionally characterized a total of 21 spruce TPSs: 12 from Sitka spruce (P. sitchensis, 5 from white spruce (P. glauca, and 4 from hybrid white spruce (P. glauca × P. engelmannii, which included 15 monoterpene synthases, 4 sesquiterpene synthases, and 2 diterpene synthases. Conclusions The functional diversity of these characterized TPSs parallels the diversity of terpenoids found in the oleoresin and volatile emissions of Sitka spruce and provides a context for understanding this chemical diversity at the molecular and mechanistic levels. The comparative characterization of Sitka spruce and Norway spruce diterpene synthases revealed the natural occurrence of TPS sequence variants between closely related spruce species, confirming a previous prediction from site-directed mutagenesis and modelling.

  2. Profiling of the Terpene Metabolome in Carrot Fruits of Wild ( Daucus carota L. ssp. carota) Accessions and Characterization of a Geraniol Synthase.

    Science.gov (United States)

    Yahyaa, Mosaab; Ibdah, Muhammad; Marzouk, Sally; Ibdah, Mwafaq

    2018-03-14

    Fruits from wild carrot ( Daucus carota L. ssp. carota) have been used for medicinal purposes since ancient times. The oil of its seeds, with their abundant monoterpenes and sesquiterpenes, has drawn attention in recent years because of its potential pharmaceutical application. A combined chemical, biochemical, and molecular study was conducted to evaluate the differential accumulation of terpene volatiles in carrot fruits of wild accessions. This work reports a similarity-based cloning strategy identification and functional characterization of one carrot monoterpene terpene synthase, WtDcTPS1. Recombinant WtDcTPS1 protein produces mainly geraniol, the predominant monoterpene in carrot seeds of wild accession 23727. The results suggest a role for the WtDcTPS1 gene in the biosynthesis of carrot fruit aroma and flavor compounds.

  3. Comparative analysis and validation of the malachite green assay for the high throughput biochemical characterization of terpene synthases.

    Science.gov (United States)

    Vardakou, Maria; Salmon, Melissa; Faraldos, Juan A; O'Maille, Paul E

    2014-01-01

    Terpenes are the largest group of natural products with important and diverse biological roles, while of tremendous economic value as fragrances, flavours and pharmaceutical agents. Class-I terpene synthases (TPSs), the dominant type of TPS enzymes, catalyze the conversion of prenyl diphosphates to often structurally diverse bioactive terpene hydrocarbons, and inorganic pyrophosphate (PPi). To measure their kinetic properties, current bio-analytical methods typically rely on the direct detection of hydrocarbon products by radioactivity measurements or gas chromatography-mass spectrometry (GC-MS). In this study we employed an established, rapid colorimetric assay, the pyrophosphate/malachite green assay (MG), as an alternative means for the biochemical characterization of class I TPSs activity.•We describe the adaptation of the MG assay for turnover and catalytic efficiency measurements of TPSs.•We validate the method by direct comparison with established assays. The agreement of k cat/K M among methods makes this adaptation optimal for rapid evaluation of TPSs.•We demonstrate the application of the MG assay for the high-throughput screening of TPS gene libraries.

  4. Natural variation in monoterpene synthesis in kiwifruit: transcriptional regulation of terpene synthases by NAC and ETHYLENE-INSENSITIVE3-like transcription factors.

    Science.gov (United States)

    Nieuwenhuizen, Niels J; Chen, Xiuyin; Wang, Mindy Y; Matich, Adam J; Perez, Ramon Lopez; Allan, Andrew C; Green, Sol A; Atkinson, Ross G

    2015-04-01

    Two kiwifruit (Actinidia) species with contrasting terpene profiles were compared to understand the regulation of fruit monoterpene production. High rates of terpinolene production in ripe Actinidia arguta fruit were correlated with increasing gene and protein expression of A. arguta terpene synthase1 (AaTPS1) and correlated with an increase in transcript levels of the 2-C-methyl-D-erythritol 4-phosphate pathway enzyme 1-deoxy-D-xylulose-5-phosphate synthase (DXS). Actinidia chinensis terpene synthase1 (AcTPS1) was identified as part of an array of eight tandemly duplicated genes, and AcTPS1 expression and terpene production were observed only at low levels in developing fruit. Transient overexpression of DXS in Nicotiana benthamiana leaves elevated monoterpene synthesis by AaTPS1 more than 100-fold, indicating that DXS is likely to be the key step in regulating 2-C-methyl-D-erythritol 4-phosphate substrate flux in kiwifruit. Comparative promoter analysis identified potential NAC (for no apical meristem [NAM], Arabidopsis transcription activation factor [ATAF], and cup-shaped cotyledon [CUC])-domain transcription factor) and ETHYLENE-INSENSITIVE3-like transcription factor (TF) binding sites in the AaTPS1 promoter, and cloned members of both TF classes were able to activate the AaTPS1 promoter in transient assays. Electrophoretic mobility shift assays showed that AaNAC2, AaNAC3, and AaNAC4 bind a 28-bp fragment of the proximal NAC binding site in the AaTPS1 promoter but not the A. chinensis AcTPS1 promoter, where the NAC binding site was mutated. Activation could be restored by reintroducing multiple repeats of the 12-bp NAC core-binding motif. The absence of NAC transcriptional activation in ripe A. chinensis fruit can account for the low accumulation of AcTPS1 transcript, protein, and monoterpene volatiles in this species. These results indicate the importance of NAC TFs in controlling monoterpene production and other traits in ripening fruits. © 2015 American

  5. Natural Variation in Monoterpene Synthesis in Kiwifruit: Transcriptional Regulation of Terpene Synthases by NAC and ETHYLENE-INSENSITIVE3-Like Transcription Factors1

    Science.gov (United States)

    Nieuwenhuizen, Niels J.; Chen, Xiuyin; Wang, Mindy Y.; Matich, Adam J.; Perez, Ramon Lopez; Allan, Andrew C.; Green, Sol A.; Atkinson, Ross G.

    2015-01-01

    Two kiwifruit (Actinidia) species with contrasting terpene profiles were compared to understand the regulation of fruit monoterpene production. High rates of terpinolene production in ripe Actinidia arguta fruit were correlated with increasing gene and protein expression of A. arguta terpene synthase1 (AaTPS1) and correlated with an increase in transcript levels of the 2-C-methyl-d-erythritol 4-phosphate pathway enzyme 1-deoxy-d-xylulose-5-phosphate synthase (DXS). Actinidia chinensis terpene synthase1 (AcTPS1) was identified as part of an array of eight tandemly duplicated genes, and AcTPS1 expression and terpene production were observed only at low levels in developing fruit. Transient overexpression of DXS in Nicotiana benthamiana leaves elevated monoterpene synthesis by AaTPS1 more than 100-fold, indicating that DXS is likely to be the key step in regulating 2-C-methyl-d-erythritol 4-phosphate substrate flux in kiwifruit. Comparative promoter analysis identified potential NAC (for no apical meristem [NAM], Arabidopsis transcription activation factor [ATAF], and cup-shaped cotyledon [CUC])-domain transcription factor) and ETHYLENE-INSENSITIVE3-like transcription factor (TF) binding sites in the AaTPS1 promoter, and cloned members of both TF classes were able to activate the AaTPS1 promoter in transient assays. Electrophoretic mobility shift assays showed that AaNAC2, AaNAC3, and AaNAC4 bind a 28-bp fragment of the proximal NAC binding site in the AaTPS1 promoter but not the A. chinensis AcTPS1 promoter, where the NAC binding site was mutated. Activation could be restored by reintroducing multiple repeats of the 12-bp NAC core-binding motif. The absence of NAC transcriptional activation in ripe A. chinensis fruit can account for the low accumulation of AcTPS1 transcript, protein, and monoterpene volatiles in this species. These results indicate the importance of NAC TFs in controlling monoterpene production and other traits in ripening fruits. PMID:25649633

  6. A high-throughput colorimetric screening assay for terpene synthase activity based on substrate consumption.

    Directory of Open Access Journals (Sweden)

    Maiko Furubayashi

    Full Text Available Terpene synthases catalyze the formation of a variety of terpene chemical structures. Systematic mutagenesis studies have been effective in providing insights into the characteristic and complex mechanisms of C-C bond formations and in exploring the enzymatic potential for inventing new chemical structures. In addition, there is growing demand to increase terpene synthase activity in heterologous hosts, given the maturation of metabolic engineering and host breeding for terpenoid synthesis. We have developed a simple screening method for the cellular activities of terpene synthases by scoring their substrate consumption based on the color loss of the cell harboring carotenoid pathways. We demonstrate that this method can be used to detect activities of various terpene synthase or prenyltransferase genes in a high-throughput manner, irrespective of the product type, enabling the mutation analysis and directed evolution of terpene synthases. We also report the possibility for substrate-specific screening system of terpene synthases by taking advantage of the substrate-size specificity of C30 and C40 carotenoid pathways.

  7. Two residues determine the product profile of the class II diterpene synthases TPS14 and TPS21 of Tripterygium wilfordii

    DEFF Research Database (Denmark)

    Hansen, Nikolaj Lervad; Nissen, Jakob N.; Hamberger, Björn Robert

    2017-01-01

    residue gave mixed product profiles. Two mutants, TwTPS14:Y265H and TwTPS21:A325V, also produced ent-copalyl diphosphate, highlighting the evolutionary potential of enzymes of this family to drive rapid diversification of plant diterpene biosynthesis through neo-functionalization. Our study contributes......The medicinal plant Tripterygium wilfordii (Celastraceae) contains a pair of class II diterpene synthases (diTPS) of specialized labdane-type metabolism that, despite remarkably close homology, form strikingly different products. TwTPS21 catalyzes bicyclization of the linear C20 precursor......-directed mutagenesis, we generated a panel of six variants, where one, or both positions were exchanged between the enzymes. In coupled heterologous assays with a corresponding class I diTPS, TwTPS2, complete product interchange was observed in variants with both reciprocal mutations, while substitutions of either...

  8. Suites of Terpene Synthases Explain Differential Terpenoid Production in Ginger and Turmeric Tissues

    Science.gov (United States)

    Koo, Hyun Jo; Gang, David R.

    2012-01-01

    The essential oils of ginger (Zingiber officinale) and turmeric (Curcuma longa) contain a large variety of terpenoids, some of which possess anticancer, antiulcer, and antioxidant properties. Despite their importance, only four terpene synthases have been identified from the Zingiberaceae family: (+)-germacrene D synthase and (S)-β-bisabolene synthase from ginger rhizome, and α-humulene synthase and β-eudesmol synthase from shampoo ginger (Zingiber zerumbet) rhizome. We report the identification of 25 mono- and 18 sesquiterpene synthases from ginger and turmeric, with 13 and 11, respectively, being functionally characterized. Novel terpene synthases, (−)-caryolan-1-ol synthase and α-zingiberene/β-sesquiphellandrene synthase, which is responsible for formation of the major sesquiterpenoids in ginger and turmeric rhizomes, were also discovered. These suites of enzymes are responsible for formation of the majority of the terpenoids present in these two plants. Structures of several were modeled, and a comparison of sets of paralogs suggests how the terpene synthases in ginger and turmeric evolved. The most abundant and most important sesquiterpenoids in turmeric rhizomes, (+)-α-turmerone and (+)-β-turmerone, are produced from (−)-α-zingiberene and (−)-β-sesquiphellandrene, respectively, via α-zingiberene/β-sesquiphellandrene oxidase and a still unidentified dehydrogenase. PMID:23272109

  9. Suites of terpene synthases explain differential terpenoid production in ginger and turmeric tissues.

    Directory of Open Access Journals (Sweden)

    Hyun Jo Koo

    Full Text Available The essential oils of ginger (Zingiber officinale and turmeric (Curcuma longa contain a large variety of terpenoids, some of which possess anticancer, antiulcer, and antioxidant properties. Despite their importance, only four terpene synthases have been identified from the Zingiberaceae family: (+-germacrene D synthase and (S-β-bisabolene synthase from ginger rhizome, and α-humulene synthase and β-eudesmol synthase from shampoo ginger (Zingiber zerumbet rhizome. We report the identification of 25 mono- and 18 sesquiterpene synthases from ginger and turmeric, with 13 and 11, respectively, being functionally characterized. Novel terpene synthases, (--caryolan-1-ol synthase and α-zingiberene/β-sesquiphellandrene synthase, which is responsible for formation of the major sesquiterpenoids in ginger and turmeric rhizomes, were also discovered. These suites of enzymes are responsible for formation of the majority of the terpenoids present in these two plants. Structures of several were modeled, and a comparison of sets of paralogs suggests how the terpene synthases in ginger and turmeric evolved. The most abundant and most important sesquiterpenoids in turmeric rhizomes, (+-α-turmerone and (+-β-turmerone, are produced from (--α-zingiberene and (--β-sesquiphellandrene, respectively, via α-zingiberene/β-sesquiphellandrene oxidase and a still unidentified dehydrogenase.

  10. Role of the Colletotrichum acutatum sesquiterpene synthase CaTPS in the biosynthesis of sesquiterpenoids

    DEFF Research Database (Denmark)

    Amby, Daniel Buchvaldt; Manczak, Tom; Petersen, Mikael Agerlin

    2016-01-01

    biosynthesis is performed by sesquiterpene synthases (TPS). Only a few TPSs have been functionally characterized from filamentous fungi and none from the genus Colletotrichum. Despite being an important fungal pathogen to agriculture, it is poorly understood at the molecular and chemical levels. The terpenoid...... characterization of TPS in Colletotrichum spp. and terpenoid profiles of Coll. acutatum, which could facilitate studies on the role of terpenoids in the ecology of Coll. acutatum....

  11. Cloning and functional characterization of three terpene synthases from lavender (Lavandula angustifolia).

    Science.gov (United States)

    Landmann, Christian; Fink, Barbara; Festner, Maria; Dregus, Márta; Engel, Karl-Heinz; Schwab, Wilfried

    2007-09-15

    The essential oil of lavender (Lavandula angustifolia) is mainly composed of mono- and sesquiterpenes. Using a homology-based PCR strategy, two monoterpene synthases (LaLIMS and LaLINS) and one sesquiterpene synthase (LaBERS) were cloned from lavender leaves and flowers. LaLIMS catalyzed the formation of (R)-(+)-limonene, terpinolene, (1R,5S)-(+)-camphene, (1R,5R)-(+)-alpha-pinene, beta-myrcene and traces of alpha-phellandrene. The proportions of these products changed significantly when Mn(2+) was supplied as the cofactor instead of Mg(2+). The second enzyme LaLINS produced exclusively (R)-(-)-linalool, the main component of lavender essential oil. LaBERS transformed farnesyl diphosphate and represents the first reported trans-alpha-bergamotene synthase. It accepted geranyl diphosphate with higher affinity than farnesyl diphosphate and also produced monoterpenes, albeit at low rates. LaBERS is probably derived from a parental monoterpene synthase by the loss of the plastidial signal peptide and by broadening its substrate acceptance spectrum. The identification and description of the first terpene synthases from L. angustifolia forms the basis for the biotechnological modification of essential oil composition in lavender.

  12. Ectopic Terpene Synthase Expression Enhances Sesquiterpene Emission in Nicotiana attenuata without Altering Defense or Development of Transgenic Plants or Neighbors1[W

    Science.gov (United States)

    Schuman, Meredith C.; Palmer-Young, Evan C.; Schmidt, Axel; Gershenzon, Jonathan; Baldwin, Ian T.

    2014-01-01

    Sesquiterpenoids, with approximately 5,000 structures, are the most diverse class of plant volatiles with manifold hypothesized functions in defense, stress tolerance, and signaling between and within plants. These hypotheses have often been tested by transforming plants with sesquiterpene synthases expressed behind the constitutively active 35S promoter, which may have physiological costs measured as inhibited growth and reduced reproduction or may require augmentation of substrate pools to achieve enhanced emission, complicating the interpretation of data from affected transgenic lines. Here, we expressed maize (Zea mays) terpene synthase10 (ZmTPS10), which produces (E)-α-bergamotene and (E)-β-farnesene, or a point mutant ZmTPS10M, which produces primarily (E)-β-farnesene, under control of the 35S promoter in the ecological model plant Nicotiana attenuata. Transgenic N. attenuata plants had specifically enhanced emission of target sesquiterpene(s) with no changes detected in their emission of any other volatiles. Treatment with herbivore or jasmonate elicitors induces emission of (E)-α-bergamotene in wild-type plants and also tended to increase emission of (E)-α-bergamotene and (E)-β-farnesene in transgenics. However, transgenics did not differ from the wild type in defense signaling or chemistry and did not alter defense chemistry in neighboring wild-type plants. These data are inconsistent with within-plant and between-plant signaling functions of (E)-β-farnesene and (E)-α-bergamotene in N. attenuata. Ectopic sesquiterpene emission was apparently not costly for transgenics, which were similar to wild-type plants in their growth and reproduction, even when forced to compete for common resources. These transgenics would be well suited for field experiments to investigate indirect ecological effects of sesquiterpenes for a wild plant in its native habitat. PMID:25187528

  13. Disruption of the Candida albicans TPS1 Gene Encoding Trehalose-6-Phosphate Synthase Impairs Formation of Hyphae and Decreases Infectivity†

    Science.gov (United States)

    Zaragoza, Oscar; Blazquez, Miguel A.; Gancedo, Carlos

    1998-01-01

    The TPS1 gene from Candida albicans, which encodes trehalose-6-phosphate synthase, has been cloned by functional complementation of a tps1 mutant from Saccharomyces cerevisiae. In contrast with the wild-type strain, the double tps1/tps1 disruptant did not accumulate trehalose at stationary phase or after heat shock. Growth of the tps1/tps1 disruptant at 30°C was indistinguishable from that of the wild type. However, at 42°C it did not grow on glucose or fructose but grew normally on galactose or glycerol. At 37°C, the yeast-hypha transition in the mutant in glucose-calf serum medium did not occur. During growth at 42°C, the mutant did not form hyphae in galactose or in glycerol. Some of the growth defects observed may be traced to an unbalanced sugar metabolism that reduces the cellular content of ATP. Mice inoculated with 106 CFU of the tps1/tps1 mutant did not show visible symptoms of infection 16 days after inoculation, while those similarly inoculated with wild-type cells were dead 12 days after inoculation. PMID:9683476

  14. The Class II trehalose 6-phosphate synthase gene PvTPS9 modulates trehalose metabolism in Phaseolus vulgaris nodules.

    Directory of Open Access Journals (Sweden)

    Aarón Barraza

    2016-11-01

    Full Text Available Legumes form symbioses with rhizobia, producing nitrogen-fixing nodules on the roots of the plant host. The network of plant signaling pathways affecting carbon metabolism may determine the final number of nodules. The trehalose biosynthetic pathway regulates carbon metabolism and plays a fundamental role in plant growth and development, as well as in plant-microbe interactions. The expression of genes for trehalose synthesis during nodule development suggests that this metabolite may play a role in legume-rhizobia symbiosis. In this work, PvTPS9, which encodes a Class II trehalose-6-phosphate synthase (TPS of common bean (Phaseolus vulgaris, was silenced by RNA interference in transgenic nodules. The silencing of PvTPS9 in root nodules resulted in a reduction of 85% (± 1% of its transcript, which correlated with a 30% decrease in trehalose contents of transgenic nodules and in untransformed leaves. Composite transgenic plants with PvTPS9 silenced in the roots showed no changes in nodule number and nitrogen fixation, but a severe reduction in plant biomass and altered transcript profiles of all Class II TPS genes. Our data suggest that PvTPS9 plays a key role in modulating trehalose metabolism in the symbiotic nodule and, therefore, in the whole plant.

  15. RNA sequencing on Solanum lycopersicum trichomes identifies transcription factors that activate terpene synthase promoters

    NARCIS (Netherlands)

    Spyropoulou, E.A.; Haring, M.A.; Schuurink, R.C.

    2014-01-01

    BACKGROUND: Glandular trichomes are production and storage organs of specialized metabolites such as terpenes, which play a role in the plant's defense system. The present study aimed to shed light on the regulation of terpene biosynthesis in Solanum lycopersicum trichomes by identification of

  16. Taxadiene Synthase Structure and Evolution of Modular Architecture in Terpene Biosynthesis

    Energy Technology Data Exchange (ETDEWEB)

    M Köksal; Y Jin; R Coates; R Croteau; D Christianson

    2011-12-31

    With more than 55,000 members identified so far in all forms of life, the family of terpene or terpenoid natural products represents the epitome of molecular biodiversity. A well-known and important member of this family is the polycyclic diterpenoid Taxol (paclitaxel), which promotes tubulin polymerization and shows remarkable efficacy in cancer chemotherapy. The first committed step of Taxol biosynthesis in the Pacific yew (Taxus brevifolia) is the cyclization of the linear isoprenoid substrate geranylgeranyl diphosphate (GGPP) to form taxa-4(5),11(12)diene, which is catalysed by taxadiene synthase. The full-length form of this diterpene cyclase contains 862 residues, but a roughly 80-residue amino-terminal transit sequence is cleaved on maturation in plastids. We now report the X-ray crystal structure of a truncation variant lacking the transit sequence and an additional 27 residues at the N terminus, hereafter designated TXS. Specifically, we have determined structures of TXS complexed with 13-aza-13,14-dihydrocopalyl diphosphate (1.82 {angstrom} resolution) and 2-fluorogeranylgeranyl diphosphate (2.25 {angstrom} resolution). The TXS structure reveals a modular assembly of three {alpha}-helical domains. The carboxy-terminal catalytic domain is a class I terpenoid cyclase, which binds and activates substrate GGPP with a three-metal ion cluster. The N-terminal domain and a third 'insertion' domain together adopt the fold of a vestigial class II terpenoid cyclase. A class II cyclase activates the isoprenoid substrate by protonation instead of ionization, and the TXS structure reveals a definitive connection between the two distinct cyclase classes in the evolution of terpenoid biosynthesis.

  17. Functional Annotation, Genome Organization and Phylogeny of the Grapevine (Vitis vinifera Terpene Synthase Gene Family Based on Genome Assembly, FLcDNA Cloning, and Enzyme Assays

    Directory of Open Access Journals (Sweden)

    Toub Omid

    2010-10-01

    Full Text Available Abstract Background Terpenoids are among the most important constituents of grape flavour and wine bouquet, and serve as useful metabolite markers in viticulture and enology. Based on the initial 8-fold sequencing of a nearly homozygous Pinot noir inbred line, 89 putative terpenoid synthase genes (VvTPS were predicted by in silico analysis of the grapevine (Vitis vinifera genome assembly 1. The finding of this very large VvTPS family, combined with the importance of terpenoid metabolism for the organoleptic properties of grapevine berries and finished wines, prompted a detailed examination of this gene family at the genomic level as well as an investigation into VvTPS biochemical functions. Results We present findings from the analysis of the up-dated 12-fold sequencing and assembly of the grapevine genome that place the number of predicted VvTPS genes at 69 putatively functional VvTPS, 20 partial VvTPS, and 63 VvTPS probable pseudogenes. Gene discovery and annotation included information about gene architecture and chromosomal location. A dense cluster of 45 VvTPS is localized on chromosome 18. Extensive FLcDNA cloning, gene synthesis, and protein expression enabled functional characterization of 39 VvTPS; this is the largest number of functionally characterized TPS for any species reported to date. Of these enzymes, 23 have unique functions and/or phylogenetic locations within the plant TPS gene family. Phylogenetic analyses of the TPS gene family showed that while most VvTPS form species-specific gene clusters, there are several examples of gene orthology with TPS of other plant species, representing perhaps more ancient VvTPS, which have maintained functions independent of speciation. Conclusions The highly expanded VvTPS gene family underpins the prominence of terpenoid metabolism in grapevine. We provide a detailed experimental functional annotation of 39 members of this important gene family in grapevine and comprehensive information

  18. The terpene synthase gene family in Tripterygium wilfordii harbors a labdane-type diterpene synthase among the monoterpene synthase TPS-b subfamily

    DEFF Research Database (Denmark)

    Hansen, Nikolaj Lervad; Heskes, Allison Maree; Hamberger, Britta

    2017-01-01

    Tripterygium wilfordii (Celastraceae) is a medicinal plant with anti-inflammatory and immunosuppressive properties. Identification of a vast array of unusual sesquiterpenoids, diterpenoids and triterpenoids in T. wilfordii has spurred investigations of their pharmacological properties. The tri-ep...

  19. Emission and Accumulation of Monoterpene and the Key Terpene Synthase (TPS) Associated with Monoterpene Biosynthesis in Osmanthus fragrans Lour

    OpenAIRE

    Zeng, Xiangling; Liu, Cai; Zheng, Riru; Cai, Xuan; Luo, Jing; Zou, Jingjing; Wang, Caiyun

    2016-01-01

    Osmanthus fragrans is an ornamental and economically important plant known for its magnificent aroma, and the most important aroma-active compounds in flowers are monoterpenes, mainly β-ocimene, linalool and linalool derivatives. To understand the molecular mechanism of monoterpene production, we analyzed the emission and accumulation patterns of these compounds and the transcript levels of the genes involved in their biosynthesis in two O. fragrans cultivars during flowering stages. The resu...

  20. Genome-wide analysis of the Solanum tuberosum (potato) trehalose-6-phosphate synthase (TPS) gene family: evolution and differential expression during development and stress.

    Science.gov (United States)

    Xu, Yingchun; Wang, Yanjie; Mattson, Neil; Yang, Liu; Jin, Qijiang

    2017-12-01

    Trehalose-6-phosphate synthase (TPS) serves important functions in plant desiccation tolerance and response to environmental stimuli. At present, a comprehensive analysis, i.e. functional classification, molecular evolution, and expression patterns of this gene family are still lacking in Solanum tuberosum (potato). In this study, a comprehensive analysis of the TPS gene family was conducted in potato. A total of eight putative potato TPS genes (StTPSs) were identified by searching the latest potato genome sequence. The amino acid identity among eight StTPSs varied from 59.91 to 89.54%. Analysis of d N /d S ratios suggested that regions in the TPP (trehalose-6-phosphate phosphatase) domains evolved faster than the TPS domains. Although the sequence of the eight StTPSs showed high similarity (2571-2796 bp), their gene length is highly differentiated (3189-8406 bp). Many of the regulatory elements possibly related to phytohormones, abiotic stress and development were identified in different TPS genes. Based on the phylogenetic tree constructed using TPS genes of potato, and four other Solanaceae plants, TPS genes could be categorized into 6 distinct groups. Analysis revealed that purifying selection most likely played a major role during the evolution of this family. Amino acid changes detected in specific branches of the phylogenetic tree suggests relaxed constraints might have contributed to functional divergence among groups. Moreover, StTPSs were found to exhibit tissue and treatment specific expression patterns upon analysis of transcriptome data, and performing qRT-PCR. This study provides a reference for genome-wide identification of the potato TPS gene family and sets a framework for further functional studies of this important gene family in development and stress response.

  1. Susceptibility to Verticillium longisporum is linked to monoterpene production by TPS23/27 in Arabidopsis.

    Science.gov (United States)

    Roos, Jonas; Bejai, Sarosh; Mozūraitis, Raimondas; Dixelius, Christina

    2015-02-01

    The fungus Verticillium longisporum is a soil-borne plant pathogen of increasing economic importance, and information on plant responses to it is limited. To identify the genes and components involved in the early stages of infection, transcripts in roots of V. longisporum-challenged Arabidopsis Col-0 and the susceptible NON-RACE SPECIFIC DISEASE RESISTANCE 1 (ndr1-1) mutant were compared using ATH1 gene chips. The analysis revealed altered transcript levels of several terpene biosynthesis genes, including the monoterpene synthase TPS23/27. When transgenic 35S:TPS23/27 and TPS23/27-amiRNA plants were monitored the over-expresser line showed enhanced fungal colonization whereas the silenced genotype was indistinguishable from Col-0. Transcript analysis of terpene biosynthesis genes suggested that only the TPS23/27 pathway is affected in the two transgenic genotypes. To confirm changes in monoterpene production, emitted volatiles were determined using solid-phase microextraction and gas chromatography-mass spectrometry. Levels of all identified TPS23/27 monoterpene products were significantly altered in the transgenic plants. A stimulatory effect on conidial germination and hyphal growth of V. longisporum was also seen in co-cultivation with 35S:TPS23/27 plants and upon exposure to 1,8-cineole, the main product of TPS23/27. Methyl jasmonate treatments of myc2-1 and myc2-2 mutants and analysis of TPS23/27:uidA in the myc2-2 background suggested a dependence on jasmonic acid mediated by the transcription factor MYC2. Taken together, our results show that TPS23/27-produced monoterpenes stimulate germination and subsequent invasion of V. longisporum in Arabidopsis roots. © 2014 The Authors The Plant Journal © 2014 John Wiley & Sons Ltd.

  2. Influence of gibberellin and daminozide on the expression of terpene synthases and on monoterpenes in common sage (Salvia officinalis).

    Science.gov (United States)

    Schmiderer, Corinna; Grausgruber-Gröger, Sabine; Grassi, Paolo; Steinborn, Ralf; Novak, Johannes

    2010-07-01

    Common sage (Salvia officinalis L., Lamiaceae) is one of the most important medicinal and aromatic plants, with antioxidant, antimicrobial, spasmolytic, astringent, antihidrotic and specific sensorial properties. The essential oil of the plant, composed mainly of the monoterpenes 1,8-cineole, alpha-thujone, beta-thujone and camphor, is responsible for some of these effects. Gibberellins regulate diverse physiological processes in plants, such as seed germination, shoot elongation and cell division. In this study, we analyzed the effect of exogenously applied plant growth regulators, namely gibberellic acid (GA(3)) and daminozide, on leaf morphology and essential oil formation of two leaf stages during the period of leaf expansion. Essential oil content increased with increasing levels of gibberellins and decreased when gibberellin biosynthesis was blocked with daminozide. With increasing levels of gibberellins, 1,8-cineole and camphor contents increased. Daminozide blocked the accumulation of alpha- and beta-thujone. GA(3) at the highest level applied also led to a significant decrease of alpha- and beta-thujone. Monoterpene synthases are a class of enzymes responsible for the first step in monoterpene biosynthesis, competing for the same substrate geranylpyrophosphate. The levels of gene expression of the three most important monoterpene synthases in sage were investigated, 1,8-cineole synthase leading directly to 1,8-cineole, (+)-sabinene synthase responsible for the first step in the formation of alpha- and beta-thujone, and (+)-bornyl diphosphate synthase, the first step in camphor biosynthesis. The foliar application of GA(3) increased, while daminozide significantly decreased gene expression of the monoterpene synthases. The amounts of two of the end products, 1,8-cineole and camphor, were directly correlated with the levels of gene expression of the respective monoterpene synthases, indicating transcriptional control, while the formation of alpha- and beta

  3. Valencene synthase from the heartwood of Nootka cypress (Callitropsis nootkatensis) for biotechnological production of valencene.

    Science.gov (United States)

    Beekwilder, Jules; van Houwelingen, Adèle; Cankar, Katarina; van Dijk, Aalt D J; de Jong, René M; Stoopen, Geert; Bouwmeester, Harro; Achkar, Jihane; Sonke, Theo; Bosch, Dirk

    2014-02-01

    Nootkatone is one of the major terpenes in the heartwood of the Nootka cypress Callitropsis nootkatensis. It is an oxidized sesquiterpene, which has been postulated to be derived from valencene. Both valencene and nootkatone are used for flavouring citrus beverages and are considered among the most valuable terpenes used at commercial scale. Functional evaluation of putative terpene synthase genes sourced by large-scale EST sequencing from Nootka cypress wood revealed a valencene synthase gene (CnVS). CnVS expression in different tissues from the tree correlates well with nootkatone content, suggesting that CnVS represents the first dedicated gene in the nootkatone biosynthetic pathway in C. nootkatensis The gene belongs to the gymnosperm-specific TPS-d subfamily of terpenes synthases and its protein sequence has low similarity to known citrus valencene synthases. In vitro, CnVS displays high robustness under different pH and temperature regimes, potentially beneficial properties for application in different host and physiological conditions. Biotechnological production of sesquiterpenes has been shown to be feasible, but productivity of microbial strains expressing valencene synthase from Citrus is low, indicating that optimization of valencene synthase activity is needed. Indeed, expression of CnVS in Saccharomyces cerevisiae indicated potential for higher yields. In an optimized Rhodobacter sphaeroides strain, expression of CnVS increased valencene yields 14-fold to 352 mg/L, bringing production to levels with industrial potential. © 2013 Society for Experimental Biology, Association of Applied Biologists and John Wiley & Sons Ltd.

  4. Characterization of two monoterpene synthases involved in floral scent formation in Hedychium coronarium.

    Science.gov (United States)

    Yue, Yuechong; Yu, Rangcai; Fan, Yanping

    2014-10-01

    Hedychium coronarium, a perennial herb belonging to the family Zingiberaceae, is cultivated as a garden plant or cut flower as well as for medicine and aromatic oil. Its flowers emit a fresh and inviting scent, which is mainly because of monoterpenes present in the profile of the floral volatiles. However, fragrance produced as a result of monoterpenes has not been well studied. In the present study, two novel terpene synthase (TPS) genes (HcTPS7 and HcTPS8) were isolated to study the biosynthesis of monoterpenes in H. coronarium. In vitro characterization showed that the recombinant HcTPS7 was capable of generating sabinene as its main product, in addition to nine sub-products from geranyl diphosphate (GPP). Recombinant HcTPS8 almost specifically catalyzed the formation of linalool from GPP, while it converted farnesyl diphosphate (FPP) to α-bergamotene, cis-α-bisabolene, β-farnesene and other ten sesquiterpenes. Subcellular localization experiments revealed that HcTPS7 and HcTPS8 were located in plastids. Real-time PCR analyses showed that HcTPS7 and HcTPS8 genes were highly expressed in petals and sepals, but were almost undetectable in vegetative organs. The changes of their expression levels in petals were positively correlated with the emission patterns of sabinene and linalool, respectively, during flower development. The results indicated that HcTPS7 and HcTPS8 were involved in the biosynthesis of sabinene and linalool in H. coronarium flowers. Results on these two TPSs first characterized from H. coronarium provide new insights into molecular mechanisms of terpene biosynthesis in this species and also lay the basis for biotechnological modification of floral scent profile in Hedychium.

  5. Methods for high yield production of terpenes

    Energy Technology Data Exchange (ETDEWEB)

    Kutchan, Toni; Higashi, Yasuhiro; Feng, Xiaohong

    2017-01-03

    Provided are enhanced high yield production systems for producing terpenes in plants via the expression of fusion proteins comprising various combinations of geranyl diphosphate synthase large and small subunits and limonene synthases. Also provided are engineered oilseed plants that accumulate monoterpene and sesquiterpene hydrocarbons in their seeds, as well as methods for producing such plants, providing a system for rapidly engineering oilseed crop production platforms for terpene-based biofuels.

  6. Insight into Biochemical Characterization of Plant Sesquiterpene Synthases

    DEFF Research Database (Denmark)

    Manczak, Tom; Simonsen, Henrik Toft

    2016-01-01

    A fast and reproducible protocol was established for enzymatic characterization of plant sesquiterpene synthases that can incorporate radioactivity in their products. The method utilizes the 96-well format in conjunction with cluster tubes and enables processing of >200 samples a day. Along...... with reduced reagent usage, it allows further reduction in the use of radioactive isotopes and flammable organic solvents. The sesquiterpene synthases previously characterized were expressed in yeast, and the plant-derived Thapsia garganica kunzeaol synthase TgTPS2 was tested in this method. KM for TgTPS2...... was found to be 0.55 μM; the turnover number, kcat, was found to be 0.29 s-1, kcat for TgTPS2 is in agreement with that of terpene synthases of other plants, and kcat/KM was found to be 0.53 s-1 μM-1 for TgTPS2. The kinetic parameters were in agreement with previously published data....

  7. Allium sativum produces terpenes with fungistatic properties in response to infection with Sclerotium cepivorum.

    Science.gov (United States)

    Pontin, Mariela; Bottini, Rubén; Burba, José Luis; Piccoli, Patricia

    2015-07-01

    This study investigated terpene biosynthesis in different tissues (root, protobulb, leaf sheath and blade) of in vitro-grown garlic plants either infected or not (control) with Sclerotium cepivorum, the causative agent of Allium White Rot disease. The terpenes identified by gas chromatography-electron impact mass spectrometry (GC-EIMS) in infected plants were nerolidol, phytol, squalene, α-pinene, terpinolene, limonene, 1,8-cineole and γ-terpinene, whose levels significantly increased when exposed to the fungus. Consistent with this, an increase in terpene synthase (TPS) activity was measured in infected plants. Among the terpenes identified, nerolidol, α-pinene and terpinolene were the most abundant with antifungal activity against S. cepivorum being assessed in vitro by mycelium growth inhibition. Nerolidol and terpinolene significantly reduced sclerotia production, while α-pinene stimulated it in a concentration-dependent manner. Parallel to fungal growth inhibition, electron microscopy observations established morphological alterations in the hyphae exposed to terpinolene and nerolidol. Differences in hyphal EtBr uptake suggested that one of the antifungal mechanisms of nerolidol and terpinolene might be disruption of fungal membrane integrity. Copyright © 2015 Elsevier Ltd. All rights reserved.

  8. Principal component analysis (PCA of volatile terpene compounds dataset emitted by genetically modified sweet orange fruits and juices in which a D-limonene synthase was either up- or down-regulated vs. empty vector controls

    Directory of Open Access Journals (Sweden)

    Ana Rodríguez

    2016-12-01

    Full Text Available We have categorized the dataset from content and emission of terpene volatiles of peel and juice in both Navelina and Pineapple sweet orange cultivars in which D-limonene was either up- (S, down-regulated (AS or non-altered (EV; control (“Impact of D-limonene synthase up- or down-regulation on sweet orange fruit and juice odor perception”(A. Rodríguez, J.E. Peris, A. Redondo, T. Shimada, E. Costell, I. Carbonell, C. Rojas, L. Peña, (2016 [1]. Data from volatile identification and quantification by HS-SPME and GC–MS were classified by Principal Component Analysis (PCA individually or as chemical groups. AS juice was characterized by the higher influence of the oxygen fraction, and S juice by the major influence of ethyl esters. S juices emitted less linalool compared to AS and EV juices.

  9. TPS for Outer Planets

    Science.gov (United States)

    Venkatapathy, Ethiraj; Ellerby, D.; Gage, P.; Gasch, M.; Hwang, H.; Prabhu, D.; Stackpoole, M.; Wercinski, Paul

    2018-01-01

    This invited talk will provide an assessment of the TPS needs for Outer Planet In-situ missions to destinations with atmosphere. The talk will outline the drivers for TPS from destination, science, mission architecture and entry environment. An assessment of the readiness of the TPS, both currently available and under development, for Saturn, Titan, Uranus and Neptune are provided. The challenges related to sustainability of the TPS for future missions are discussed.

  10. Isolation and functional characterization of a τ-cadinol synthase, a new sesquiterpene synthase from Lavandula angustifolia.

    Science.gov (United States)

    Jullien, Frédéric; Moja, Sandrine; Bony, Aurélie; Legrand, Sylvain; Petit, Cécile; Benabdelkader, Tarek; Poirot, Kévin; Fiorucci, Sébastien; Guitton, Yann; Nicolè, Florence; Baudino, Sylvie; Magnard, Jean-Louis

    2014-01-01

    In this paper we characterize three sTPSs: a germacrene D (LaGERDS), a (E)-β-caryophyllene (LaCARS) and a τ-cadinol synthase (LaCADS). τ-cadinol synthase is reported here for the first time and its activity was studied in several biological models including transiently or stably transformed tobacco species. Three dimensional structure models of LaCADS and Ocimum basilicum γ-cadinene synthase were built by homology modeling using the template structure of Gossypium arboreum δ-cadinene synthase. The depiction of their active site organization provides evidence of the global influence of the enzymes on the formation of τ-cadinol: instead of a unique amino-acid, the electrostatic properties and solvent accessibility of the whole active site in LaCADS may explain the stabilization of the cadinyl cation intermediate. Quantitative PCR performed from leaves and inflorescences showed two patterns of expression. LaGERDS and LaCARS were mainly expressed during early stages of flower development and, at these stages, transcript levels paralleled the accumulation of the corresponding terpene products (germacrene D and (E)-β-caryophyllene). By contrast, the expression level of LaCADS was constant in leaves and flowers. Phylogenetic analysis provided informative results on potential duplication process leading to sTPS diversification in lavender.

  11. [Regulation of terpene metabolism

    Energy Technology Data Exchange (ETDEWEB)

    Croteau, R.

    1989-11-09

    Terpenoid oils, resins, and waxes from plants are important renewable resources. The objective of this project is to understand the regulation of terpenoid metabolism using the monoterpenes (C[sub 10]) as a model. The pathways of monoterpene biosynthesis and catabolism have been established, and the relevant enzymes characterized. Developmental studies relating enzyme levels to terpene accumulation within the oil gland sites of synthesis, and work with bioregulators, indicate that monoterpene production is controlled by terpene cyclases, the enzymes catalyzing the first step of the monoterpene pathway. As the leaf oil glands mature, cyclase levels decline and monoterpene biosynthesis ceases. Yield then decreases as the monoterpenes undergo catabolism by a process involving conversion to a glycoside and transport from the leaf glands to the root. At this site, the terpenoid is oxidatively degraded to acetate that is recycled into other lipid metabolites. During the transition from terpene biosynthesis to catabolism, the oil glands undergo dramatic ultrastructural modification. Degradation of the producing cells results in mixing of previously compartmentized monoterpenes with the catabolic enzymes, ultimately leading to yield decline. This regulatory model is being applied to the formation of other terpenoid classes (C[sub 15] C[sub 20], C[sub 30], C[sub 40]) within the oil glands. Preliminary investigations on the formation of sesquiterpenes (C[sub 15]) suggest that the corresponding cyclases may play a lesser role in determining yield of these products, but that compartmentation effects are important. From these studies, a comprehensive scheme for the regulation of terpene metabolism is being constructed. Results from this project wail have important consequences for the yield and composition of terpenoid natural products that can be made available for industrial exploitation.

  12. Cloning and functional characterization of β-phellandrene synthase from Lavandula angustifolia.

    Science.gov (United States)

    Demissie, Zerihun A; Sarker, Lukman S; Mahmoud, Soheil S

    2011-04-01

    En route to building genomics resources for Lavandula, we have obtained over 14,000 ESTs for leaves and flowers of L. angustifolia, a major essential oil crop, and identified a number of previously uncharacterized terpene synthase (TPS) genes. Here we report the cloning, expression in E. coli, and functional characterization of β-phellandrene synthase, LaβPHLS. The ORF--excluding the transit peptide--for this gene encoded a 62.3 kDa protein that contained all conserved motifs present in plant TPSs. Expression in bacteria resulted in the production of a soluble protein that was purified by Ni-NTA agarose affinity chromatography. While the recombinant LaβPHLS did not utilize FPP as a substrate, it converted GPP (the preferred substrate) and NPP into β-phellandrene as the major product, with K (m) and k (cat) of 6.55 μM and 1.75 × 10(-2) s(-1), respectively, for GPP. The LaβPHLS transcripts were highly abundant in young leaves where β-phellandrene is produced, but were barely detectable in flowers and older leaves, where β-phellandrene is not synthesized in significant quantities. This data indicate that β-phellandrene biosynthesis is transcriptionally and developmentally regulated. We also cloned and expressed in E. coli a second TPS-like protein, LaTPS-I, that lacks an internal stretch of 73 amino acids, including the signature DDxxD divalent metal binding motif, compared to other plant TPSs. The recombinant LaTPS-I did not produce detectable products in vitro when assayed with GPP, NPP or FPP as substrates. The lack of activity is most likely due to the absence of catalytically important amino acid residues within the missing region.

  13. [Regulation of terpene metabolism

    Energy Technology Data Exchange (ETDEWEB)

    Croteau, R.

    1992-01-01

    This report describes accomplishments over the past year on understanding of terpene synthesis in mint plants and sage. Specifically reported are the fractionation of 4-S-limonene synthetase, the enzyme responsible for the first committed step to monoterpene synthesis, along with isolation of the corresponding RNA and DNA cloning of its gene; the localization of the enzyme within the oil glands, regulation of transcription and translation of the synthetase, the pathway to camphor biosynthesis,a nd studies on the early stages and branch points of the isoprenoid pathway.

  14. Test Program Set (TPS) Lab

    Data.gov (United States)

    Federal Laboratory Consortium — The ARDEC TPS Laboratory provides an organic Test Program Set (TPS) development, maintenance, and life cycle management capability for DoD LCMC materiel developers....

  15. Transcriptome resources and functional characterization of monoterpene synthases for two host species of the mountain pine beetle, lodgepole pine (Pinus contorta) and jack pine (Pinus banksiana)

    Science.gov (United States)

    2013-01-01

    Background The mountain pine beetle (MPB, Dendroctonus ponderosae) epidemic has affected lodgepole pine (Pinus contorta) across an area of more than 18 million hectares of pine forests in western Canada, and is a threat to the boreal jack pine (Pinus banksiana) forest. Defence of pines against MPB and associated fungal pathogens, as well as other pests, involves oleoresin monoterpenes, which are biosynthesized by families of terpene synthases (TPSs). Volatile monoterpenes also serve as host recognition cues for MPB and as precursors for MPB pheromones. The genes responsible for terpene biosynthesis in jack pine and lodgepole pine were previously unknown. Results We report the generation and quality assessment of assembled transcriptome resources for lodgepole pine and jack pine using Sanger, Roche 454, and Illumina sequencing technologies. Assemblies revealed transcripts for approximately 20,000 - 30,000 genes from each species and assembly analyses led to the identification of candidate full-length prenyl transferase, TPS, and P450 genes of oleoresin biosynthesis. We cloned and functionally characterized, via expression of recombinant proteins in E. coli, nine different jack pine and eight different lodgepole pine mono-TPSs. The newly identified lodgepole pine and jack pine mono-TPSs include (+)-α-pinene synthases, (-)-α-pinene synthases, (-)-β-pinene synthases, (+)-3-carene synthases, and (-)-β-phellandrene synthases from each of the two species. Conclusion In the absence of genome sequences, transcriptome assemblies are important for defence gene discovery in lodgepole pine and jack pine, as demonstrated here for the terpenoid pathway genes. The product profiles of the functionally annotated mono-TPSs described here can account for the major monoterpene metabolites identified in lodgepole pine and jack pine. PMID:23679205

  16. Transcriptome resources and functional characterization of monoterpene synthases for two host species of the mountain pine beetle, lodgepole pine (Pinus contorta) and jack pine (Pinus banksiana).

    Science.gov (United States)

    Hall, Dawn E; Yuen, Macaire M S; Jancsik, Sharon; Quesada, Alfonso Lara; Dullat, Harpreet K; Li, Maria; Henderson, Hannah; Arango-Velez, Adriana; Liao, Nancy Y; Docking, Roderick T; Chan, Simon K; Cooke, Janice Ek; Breuil, Colette; Jones, Steven Jm; Keeling, Christopher I; Bohlmann, Jörg

    2013-05-16

    The mountain pine beetle (MPB, Dendroctonus ponderosae) epidemic has affected lodgepole pine (Pinus contorta) across an area of more than 18 million hectares of pine forests in western Canada, and is a threat to the boreal jack pine (Pinus banksiana) forest. Defence of pines against MPB and associated fungal pathogens, as well as other pests, involves oleoresin monoterpenes, which are biosynthesized by families of terpene synthases (TPSs). Volatile monoterpenes also serve as host recognition cues for MPB and as precursors for MPB pheromones. The genes responsible for terpene biosynthesis in jack pine and lodgepole pine were previously unknown. We report the generation and quality assessment of assembled transcriptome resources for lodgepole pine and jack pine using Sanger, Roche 454, and Illumina sequencing technologies. Assemblies revealed transcripts for approximately 20,000 - 30,000 genes from each species and assembly analyses led to the identification of candidate full-length prenyl transferase, TPS, and P450 genes of oleoresin biosynthesis. We cloned and functionally characterized, via expression of recombinant proteins in E. coli, nine different jack pine and eight different lodgepole pine mono-TPSs. The newly identified lodgepole pine and jack pine mono-TPSs include (+)-α-pinene synthases, (-)-α-pinene synthases, (-)-β-pinene synthases, (+)-3-carene synthases, and (-)-β-phellandrene synthases from each of the two species. In the absence of genome sequences, transcriptome assemblies are important for defence gene discovery in lodgepole pine and jack pine, as demonstrated here for the terpenoid pathway genes. The product profiles of the functionally annotated mono-TPSs described here can account for the major monoterpene metabolites identified in lodgepole pine and jack pine.

  17. Structural Basis of Catalysis in the Bacterial Monoterpene Synthases Linalool Synthase and 1,8-Cineole Synthase

    OpenAIRE

    Karuppiah, Vijaykumar; Ranaghan, Kara E.; Leferink, Nicole G. H.; Johannissen, Linus O.; Shanmugam, Muralidharan; Ní Cheallaigh, Aisling; Bennett, Nathan J.; Kearsey, Lewis J.; Takano, Eriko; Gardiner, John M.; van der Kamp, Marc W.; Hay, Sam; Mulholland, Adrian J.; Leys, David; Scrutton, Nigel S.

    2017-01-01

    Terpenoids form the largest and stereochemically most diverse class of natural products, and there is considerable interest in producing these by biocatalysis with whole cells or purified enzymes, and by metabolic engineering. The monoterpenes are an important class of terpenes and are industrially important as flavors and fragrances. We report here structures for the recently discovered Streptomyces clavuligerus monoterpene synthases linalool synthase (bLinS) and 1,8-cineole synthase (bCinS)...

  18. [Regulation of terpene metabolism

    Energy Technology Data Exchange (ETDEWEB)

    Croteau, R.

    1991-01-01

    During the last grant period, we have completed studies on the key pathways of monoterpene biosynthesis and catabolism in sage and peppermint, and have, by several lines of evidence, deciphered the rate-limiting step of each pathway. We have at least partially purified and characterized the relevant enzymes of each pathway. We have made a strong case, based on analytical, in vivo, and in vitro studies, that terpene accumulation depends upon the balance between biosynthesis and catabolism, and provided supporting evidence that these processes are developmentally-regulated and very closely associated with senescence of the oil glands. Oil gland ontogeny has been characterized at the ultrastructural level. We have exploited foliar-applied bioregulators to delay gland senescence, and have developed tissue explant and cell culture systems to study several elusive aspects of catabolism. We have isolated pure gland cell clusters and localized monoterpene biosynthesis and catabolism within these structures, and have used these preparations as starting materials for the purification to homogeneity of target regulatory'' enzymes. We have thus developed the necessary background knowledge, based on a firm understanding of enzymology, as well as the necessary experimental tools for studying the regulation of monoterpene metabolism at the molecular level. Furthermore, we are now in a position to extend our systematic approach to other terpenoid classes (C[sub 15]-C[sub 30]) produced by oil glands.

  19. Bornyl-diphosphate synthase from Lavandula angustifolia: A major monoterpene synthase involved in essential oil quality.

    Science.gov (United States)

    Despinasse, Yolande; Fiorucci, Sébastien; Antonczak, Serge; Moja, Sandrine; Bony, Aurélie; Nicolè, Florence; Baudino, Sylvie; Magnard, Jean-Louis; Jullien, Frédéric

    2017-05-01

    Lavender essential oils (EOs) of higher quality are produced by a few Lavandula angustifolia cultivars and mainly used in the perfume industry. Undesirable compounds such as camphor and borneol are also synthesized by lavender leading to a depreciated EO. Here, we report the cloning of bornyl diphosphate synthase of lavender (LaBPPS), an enzyme that catalyzes the production of bornyl diphosphate (BPP) and then by-products such as borneol or camphor, from an EST library. Compared to the BPPS of Salvia officinalis, the functional characterization of LaBPPS showed several differences in amino acid sequence, and the distribution of catalyzed products. Molecular modeling of the enzyme's active site suggests that the carbocation intermediates are more stable in LaBPPS than in SoBPPS leading probably to a lower efficiency of LaBPPS to convert GPP into BPP. Quantitative RT-PCR performed from leaves and flowers at different development stages of L. angustifolia samples show a clear correlation between transcript level of LaBPPS and accumulation of borneol/camphor, suggesting that LaBPPS is mainly responsible of in vivo biosynthesis of borneol/camphor in fine lavender. A phylogenetic analysis of terpene synthases (TPS) pointed out the basal position of LaBPPS in the TPSb clade, suggesting that LaBPPS could be an ancestor of others lavender TPSb. Finally, borneol could be one of the first monoterpenes to be synthesized in the Lavandula subgenus. Knowledge gained from these experiments will facilitate future studies to improve the lavender oils through metabolic engineering or plant breeding. Accession numbers: LaBPPS: KM015221. Copyright © 2017. Published by Elsevier Ltd.

  20. Regulation of terpene metabolism. Progress report, 1983

    International Nuclear Information System (INIS)

    Croteau, R.

    1986-01-01

    Studies on the metabolism of terpenes by peppermint (Menta piperita) are described. The studies describe the characterization of enzymes involved in the biosynthesis and catabolism of terpenes and the ultrastructure of the oil glands. 10 refs. (DT)

  1. Isolation and characterization of three new monoterpene synthases from Artemisia annua

    OpenAIRE

    Ju-Xin eRuan; Jian-Xu eLi; Xin eFang; Ling-Jian eWang; Wen-Li eHu; Xiao-Ya eChen; Changqing eYang

    2016-01-01

    Artemisia annua, an annual herb used in traditional Chinese medicine, produces a wealth of monoterpenes and sesquiterpenes, including the well-known sesquiterpene lactone artemisinin, an active ingredient in the treatment for malaria. Here we report three new monoterpene synthases of A. annua. From a glandular trichome cDNA library, monoterpene synthases of AaTPS2, AaTPS5 and AaTPS6, were isolated and characterized. The recombinant proteins of AaTPS5 and AaTPS6 produced multiple products with...

  2. Cloning and Comparative Studies of Seaweed Trehalose-6-Phosphate Synthase Genes

    Directory of Open Access Journals (Sweden)

    Bin Wang

    2010-07-01

    Full Text Available The full-length cDNA sequence (3219 base pairs of the trehalose-6-phosphate synthase gene of Porphyra yezoensis (PyTPS was isolated byRACE-PCR and deposited in GenBank (NCBI with the accession number AY729671. PyTPS encodes a protein of 908 amino acids before a stop codon, and has a calculated molecular mass of 101,591 Daltons. The PyTPS protein consists of a TPS domain in the N-terminus and a putative TPP domain at the C-terminus. Homology alignment for PyTPS and the TPS proteins from bacteria, yeast and higher plants indicated that the most closely related sequences to PyTPS were those from higher plants (OsTPS and AtTPS5, whereas the most distant sequence to PyTPS was from bacteria (EcOtsAB. Based on the identified sequence of the PyTPS gene, PCR primers were designed and used to amplify the TPS genes from nine other seaweed species. Sequences of the nine obtained TPS genes were deposited in GenBank (NCBI. All 10 TPS genes encoded peptides of 908 amino acids and the sequences were highly conserved both in nucleotide composition (>94% and in amino acid composition (>96%. Unlike the TPS genes from some other plants, there was no intron in any of the 10 isolated seaweed TPS genes.

  3. Isolation and Characterization of Three New Monoterpene Synthases from Artemisia annua

    Science.gov (United States)

    Ruan, Ju-Xin; Li, Jian-Xu; Fang, Xin; Wang, Ling-Jian; Hu, Wen-Li; Chen, Xiao-Ya; Yang, Chang-Qing

    2016-01-01

    Artemisia annua, an annual herb used in traditional Chinese medicine, produces a wealth of monoterpenes and sesquiterpenes, including the well-known sesquiterpene lactone artemisinin, an active ingredient in the treatment for malaria. Here we report three new monoterpene synthases of A. annua. From a glandular trichome cDNA library, monoterpene synthases of AaTPS2, AaTPS5, and AaTPS6, were isolated and characterized. The recombinant proteins of AaTPS5 and AaTPS6 produced multiple products with camphene and 1,8-cineole as major products, respectively, and AaTPS2 produced a single product, β-myrcene. Although both Mg2+ and Mn2+ were able to support their catalytic activities, altered product spectrum was observed in the presence of Mn2+ for AaTPS2 and AaTPS5. Analysis of extracts of aerial tissues and root of A. annua with gas chromatography–mass spectrometry detected more than 20 monoterpenes, of which the three enzymes constituted more than 1/3 of the total. Mechanical wounding induced the expression of all three monoterpene synthase genes, and transcript levels of AaTPS5 and AaTPS6 were also elevated after treatments with phytohormones of methyl jasmonate, salicylic acid, and gibberellin, suggesting a role of these monoterpene synthases in plant–environment interactions. The three new monoterpene synthases reported here further our understanding of molecular basis of monoterpene biosynthesis and regulation in plant. PMID:27242840

  4. Phosphorylation and 14-3-3 binding of Arabidopsis trehalose-phosphate synthase 5 in response to 2-deoxyglucose

    DEFF Research Database (Denmark)

    Harthill, Jean E; Meek, Sarah E M; Morrice, Nick

    2006-01-01

    Trehalose-6-phosphate is a 'sugar signal' that regulates plant metabolism and development. The Arabidopsis genome encodes trehalose-6-phosphate synthase (TPS) and trehalose-6-phosphatase (TPP) enzymes. It also encodes class II proteins (TPS isoforms 5-11) that contain both TPS-like and TPP...

  5. Engineering Escherichia coli for the production of terpene mixture enriched in caryophyllene and caryophyllene alcohol as potential aviation fuel compounds

    Directory of Open Access Journals (Sweden)

    Weihua Wu

    2018-06-01

    Full Text Available Recent studies have revealed that caryophyllene and its stereoisomers not only exhibit multiple biological activities but also have desired properties as renewable candidates for ground transportation and jet fuel applications. This study presents the first significant production of caryophyllene and caryolan-1-ol by an engineered E. coli with heterologous expression of mevalonate pathway genes with a caryophyllene synthase and a caryolan-1-ol synthase. By optimizing metabolic flux and fermentation parameters, the engineered strains yielded 449 mg/L of total terpene, including 406 mg/L sesquiterpene with 100 mg/L caryophyllene and 10 mg/L caryolan-1-ol. Furthermore, a marine microalgae hydrolysate was used as the sole carbon source for the production of caryophyllene and other terpene compounds. Under the optimal fermentation conditions, 360 mg/L of total terpene, 322 mg/L of sesquiterpene, and 75 mg/L caryophyllene were obtained from the pretreated algae hydrolysates. The highest yields achieved on the biomass basis were 48 mg total terpene/g algae and 10 mg caryophyllene/g algae and the caryophyllene yield is approximately ten times higher than that from plant tissues by solvent extraction. The study provides a sustainable alternative for production of caryophyllene and its alcohol from microalgae biomass as potential candidates for next generation aviation fuels. Keywords: Caryophyllene, Caryolan-1-ol, Caryophyllene synthase, Caryolan-1-ol synthase, Mevalonate pathway, Bioproduct

  6. Identification and Functional Characterization of Sesquiterpene Synthases from Xanthium strumarium.

    Science.gov (United States)

    Li, Yuanjun; Chen, Fangfang; Li, Zhenqiu; Li, Changfu; Zhang, Yansheng

    2016-03-01

    Xanthium strumarium synthesizes various pharmacologically active sesquiterpenes. The molecular characterization of sesquiterpene biosynthesis in X. strumarium has not been reported so far. In this study, the cDNAs coding for three sesquiterpene synthases (designated as XsTPS1, XsTPS2 and XsTPS3) were isolated using the X. strumarium transcriptome that we recently constructed. XsTPS1, XsTPS2 and XsTPS3 were revealed to have primary activities forming germacrene D, guaia-4,6-diene and germacrene A, respectively, by either ectopic expression in yeast cells or purified recombinant protein-based in vitro assays. Quantitative real-time PCRs and metabolite analysis for the different plant parts showed that the transcript abundance of XsTPS1-XsTPS3 is consistent with the accumulation pattern of their enzymatic products, supporting their biochemical functions in vivo. In particular, we discovered that none of the XsTPS2 product, guaia-4,6-diene, can be detected in one of the X. strumarium cultivars used in this study (it was named the Hubei-cultivar), in which a natural deletion of two A bases in the XsTPS2 cDNA disrupts its activity, which further confirmed the proposed biochemical role of XsTPS2 in X. strumarium in vivo. © The Author 2016. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oup.com.

  7. New TPS materials for aerocapture

    International Nuclear Information System (INIS)

    Laub, Bernard

    2002-01-01

    Many planetary probes, landers and aerocapture concepts are conceived for entry trajectories where peak convective heat flux is in the range 150-400 W/cm2. This may be too severe an environment for either reusable or low-density ablative materials. The high-density ablatives will work in such environments but the associated TPS weight requirements can be prohibitive. Unfortunately, there are few, if any, well-understood materials that provide reliable, predictable ablative performance for the 150-400 W/cm2 regime while still providing weight efficient TPS solutions. JPL has recently been evaluating an Earth aerocapture demonstration at an entry velocity of ≅10 km/s. TPS thickness and areal weight requirements were determined for current ablative TPS candidates (e.g., SLA-561V, PICA) where, for the large integrated heat loads associated with aerocapture, it is shown that some of these materials may not provide efficient thermal protection. A new concept, employing a low catalycity, high emissivity coating on a low-density ceramic tile is evaluated and shown to provide significant benefits for such missions

  8. Transformation of trehalose synthase gene (TPS Gene) into corn ...

    African Journals Online (AJOL)

    ARL

    2011-11-02

    Nov 2, 2011 ... With water resources increasingly diminishing, drought has become the upmost .... six to seven leaf stage, DNA was extracted for molecule detection and the .... condition of drought stress, and their electrolyte leakage rate was ...

  9. Identification of a Fungal 1,8-Cineole Synthase from Hypoxylon sp. with Specificity Determinants in Common with the Plant Synthases*

    Science.gov (United States)

    Shaw, Jeffrey J.; Berbasova, Tetyana; Sasaki, Tomoaki; Jefferson-George, Kyra; Spakowicz, Daniel J.; Dunican, Brian F.; Portero, Carolina E.; Narváez-Trujillo, Alexandra; Strobel, Scott A.

    2015-01-01

    Terpenes are an important and diverse class of secondary metabolites widely produced by fungi. Volatile compound screening of a fungal endophyte collection revealed a number of isolates in the family Xylariaceae, producing a series of terpene molecules, including 1,8-cineole. This compound is a commercially important component of eucalyptus oil used in pharmaceutical applications and has been explored as a potential biofuel additive. The genes that produce terpene molecules, such as 1,8-cineole, have been little explored in fungi, providing an opportunity to explore the biosynthetic origin of these compounds. Through genome sequencing of cineole-producing isolate E7406B, we were able to identify 11 new terpene synthase genes. Expressing a subset of these genes in Escherichia coli allowed identification of the hyp3 gene, responsible for 1,8-cineole biosynthesis, the first monoterpene synthase discovered in fungi. In a striking example of convergent evolution, mutational analysis of this terpene synthase revealed an active site asparagine critical for water capture and specificity during cineole synthesis, the same mechanism used in an unrelated plant homologue. These studies have provided insight into the evolutionary relationship of fungal terpene synthases to those in plants and bacteria and further established fungi as a relatively untapped source of this important and diverse class of compounds. PMID:25648891

  10. Isolation and characterization of three new monoterpene synthases from Artemisia annua

    Directory of Open Access Journals (Sweden)

    Ju-Xin eRuan

    2016-05-01

    Full Text Available Artemisia annua, an annual herb used in traditional Chinese medicine, produces a wealth of monoterpenes and sesquiterpenes, including the well-known sesquiterpene lactone artemisinin, an active ingredient in the treatment for malaria. Here we report three new monoterpene synthases of A. annua. From a glandular trichome cDNA library, monoterpene synthases of AaTPS2, AaTPS5 and AaTPS6, were isolated and characterized. The recombinant proteins of AaTPS5 and AaTPS6 produced multiple products with camphene and 1,8-cineole as major products, respectively, and AaTPS2 produced a single product, β-myrcene. Although both Mg2+ and Mn2+ were able to support their catalytic activities, altered product spectrum was observed in the presence of Mn2+ for AaTPS2 and AaTPS5. Analysis of extracts of aerial tissues and root of A. annua with gas chromatography-mass spectrometry (GC-MS detected more than 20 monoterpenes, of which the three enzymes constituted more than 1/3 of the total. Mechanical wounding induced the expression of all three monoterpene synthase genes, and transcript levels of AaTPS5 and AaTPS6 were also elevated after treatments with phytohormones of methyl jasmonate (MeJA, salicylic acid (SA and gibberellin (GA, suggesting a role of these monoterpene synthases in plant-environment interactions. The three new monoterpene synthases reported here further our understanding of molecular basis of monoterpene biosynthesis and regulation in plant.

  11. Simultaneous Determination of Flavonols and Terpene Lactones in ...

    African Journals Online (AJOL)

    Liquid Chromatography-Tandem - Mass Spectrometry: 1. ... Results: The method showed high selectivity of the flavonols and terpene ... Lower limit of quantification (LLOQ) was 1.232, 0.240, 0.200, ... flavonoid glycosides and terpene lactones.

  12. 40 CFR 721.9635 - Terpene residue distillates.

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 30 2010-07-01 2010-07-01 false Terpene residue distillates. 721.9635... Substances § 721.9635 Terpene residue distillates. (a) Chemical substance and significant new uses subject to reporting. (1) The chemical substance identified generically as terpene residue distillates (PMN P-96-897...

  13. Diterpene synthases of the biosynthetic system of medicinally active diterpenoids in Marrubium vulgare

    DEFF Research Database (Denmark)

    Zerbe, Philipp; Chiang, Angela; Dullat, Harpreet

    2014-01-01

    Marrubium vulgare (Lamiaceae) is a medicinal plant whose major bioactive compounds, marrubiin and other labdane-related furanoid diterpenoids, have potential applications as anti-diabetics, analgesics or vasorelaxants. Metabolite and transcriptome profiling of M. vulgare leaves identified five...... different candidate diterpene synthases (diTPSs) of the TPS-c and TPS-e/f clades. We describe the in vitro and in vivo functional characterization of the M. vulgare diTPS family. In addition to MvEKS ent-kaurene synthase of general metabolism, we identified three diTPSs of specialized metabolism: MvCPS3...

  14. BIOMIMETIC STRATEGIES IN ORGANIC SYNTHESIS. TERPENES

    Directory of Open Access Journals (Sweden)

    V. Kulcitki

    2012-12-01

    Full Text Available The current paper represents an outline of the selected contributions to the biomimetic procedures and approaches for the synthesis of terpenes with complex structure and diverse functionalisation pattern. These include homologation strategies, cyclisations, rearrangements, as well as biomimetic remote functionalisations.

  15. Identification of TPS family members in apple (Malus x domestica Borkh.) and the effect of sucrose sprays on TPS expression and floral induction.

    Science.gov (United States)

    Du, Lisha; Qi, Siyan; Ma, Juanjuan; Xing, Libo; Fan, Sheng; Zhang, Songwen; Li, Youmei; Shen, Yawen; Zhang, Dong; Han, Mingyu

    2017-11-01

    Trehalose (α-D-glucopyranosyl α-D-glucopyranoside) is a non-reducing disaccharide that serves as a carbon source and stress protectant in apple trees. Trehalose-6-phosphate (T6P) is the biosynthetic precursor of trehalose. It functions as a crucial signaling molecule involved in the regulation of floral induction, and is closely related to sucrose. Trehalose-6-phosphate synthase (TPS) family members are pivotal components of the T6P biosynthetic pathway. The present study identified 13 apple TPS family members and characterized their expression patterns in different tissues and in response to exogenous application of sucrose during floral induction. 'Fuji' apple trees were sprayed with sucrose prior to the onset of floral induction. Bud growth, flowering rate, and endogenous sugar levels were then monitored. The expression of genes associated with sucrose metabolism and flowering were also characterized by RT-quantitative PCR. Results revealed that sucrose applications significantly improved flower production and increased bud size and fresh weight, as well as the sucrose content in buds and leaves. Furthermore, the expression of MdTPS1, 2, 4, 10, and 11 was rapidly and significantly up-regulated in response to the sucrose treatments. In addition, the expression levels of flowering-related genes (e.g., SPL genes, FT1, and AP1) also increased in response to the sucrose sprays. In summary, apple TPS family members were identified that may influence the regulation of floral induction and other responses to sucrose. The relationship between sucrose and T6P or TPS during the regulation of floral induction in apple trees is discussed. Copyright © 2017 Elsevier Masson SAS. All rights reserved.

  16. Differential sensing using proteins: exploiting the cross-reactivity of serum albumin to pattern individual terpenes and terpenes in perfume.

    Science.gov (United States)

    Adams, Michelle M; Anslyn, Eric V

    2009-12-02

    There has been a growing interest in the use of differential sensing for analyte classification. In an effort to mimic the mammalian senses of taste and smell, which utilize protein-based receptors, we have introduced serum albumins as nonselective receptors for recognition of small hydrophobic molecules. Herein, we employ a sensing ensemble consisting of serum albumins, a hydrophobic fluorescent indicator (PRODAN), and a hydrophobic additive (deoxycholate) to detect terpenes. With the aid of linear discriminant analysis, we successfully applied our system to differentiate five terpenes. We then extended our terpene analysis and utilized our sensing ensemble for terpene discrimination within the complex mixtures found in perfume.

  17. Biosynthesis of the oxygenated diterpene nezukol in the medicinal plant Isodon rubescens is catalyzed by a pair of diterpene synthases.

    Directory of Open Access Journals (Sweden)

    Kyle A Pelot

    Full Text Available Plants produce an immense diversity of natural products (i.e. secondary or specialized metabolites that offer a rich source of known and potentially new pharmaceuticals and other desirable bioproducts. The Traditional Chinese Medicinal plant Isodon rubescens (Lamiaceae contains an array of bioactive labdane-related diterpenoid natural products. Of these, the ent-kauranoid oridonin is the most prominent specialized metabolite that has been extensively studied for its potent antimicrobial and anticancer efficacy. Mining of a previously established transcriptome of I. rubescens leaf tissue identified seven diterpene synthase (diTPSs candidates. Here we report the functional characterization of four I. rubescens diTPSs. IrTPS5 and IrTPS3 were identified as an ent-copalyl diphosphate (CPP synthase and a (+-CPP synthase, respectively. Distinct transcript abundance of IrTPS5 and the predicted ent-CPP synthase IrTPS1 suggested a role of IrTPS5 in specialized ent-kaurene metabolism possibly en route to oridonin. Nicotiana benthamiana co-expression assays demonstrated that IrTPS4 functions sequentially with IrTPS3 to form miltiradiene. In addition, IrTPS2 converted the IrTPS3 product (+-CPP into the hydroxylated tricyclic diterpene nezukol not previously identified in I. rubescens. Metabolite profiling verified the presence of nezukol in I. rubescens leaf tissue. The proposed IrTPS2-catalyzed reaction mechanism proceeds via the common ionization of the diphosphate group of (+-CPP, followed by formation of an intermediary pimar-15-en-8-yl+ carbocation and neutralization of the carbocation by water capture at C-8 to yield nezukol, as confirmed by nuclear magnetic resonance (NMR analysis. Oxygenation activity is rare for the family of class I diTPSs and offers new catalysts for developing metabolic engineering platforms to produce a broader spectrum of bioactive diterpenoid natural products.

  18. [Regulation of terpene metabolism.] Progress report

    International Nuclear Information System (INIS)

    Croteau, R.

    1984-01-01

    This research program represents a very broad-based approach to understanding the biochemistry of the monoterpene and sesquiterpene constituents of the essential oils. This program includes basic research on the pathways, enzymes and mechanisms of terpene biosynthesis and catabolism, on the physiology of essential oil production, and on the morphology and development of oil glands, as well as practical approaches to manipulating essential oil composition and yield. As a natural extension of research on monoterpene biosynthesis and catabolism in sage and peppermint we have explored some aspects of possible regulatory mechanisms. Tentative evidence has been obtained for developmental regulation of the levels of biosynthetic and catabolic enzymes. 10 refs., 8 figs

  19. Engineering a Synthetic Microbial Consortium for Comprehensive Conversion of Algae Biomass into Terpenes for Advanced Biofuels and Bioproducts

    Energy Technology Data Exchange (ETDEWEB)

    Wu, Weihua [Sandia National Lab. (SNL-CA), Livermore, CA (United States); Wu, Benjamin Chiau-Pin [Sandia National Lab. (SNL-CA), Livermore, CA (United States); Davis, Ryan Wesley [Sandia National Lab. (SNL-CA), Livermore, CA (United States)

    2015-10-01

    Recent strategies for algae-based biofuels have primarily focused on biodiesel production by exploiting high algal lipid yields under nutrient stress conditions. However, under conditions supporting robust algal biomass accumulation, carbohydrate and proteins typically comprise up to ~80% of the ash-free dry weight of algae biomass. Therefore, comprehensive utilization of algal biomass for production of multipurpose intermediate- to high-value bio-based products will promote scale-up of algae production and processing to commodity volumes. Terpenes are hydrocarbon and hydrocarbon-like (C:O>10:1) compounds with high energy density, and are therefore potentially promising candidates for the next generation of value added bio-based chemicals and “drop-in” replacements for petroleum-based fuels. In this study, we demonstrated the feasibility of bioconversion of proteins into sesquiterpene compounds as well as comprehensive bioconversion of algal carbohydrates and proteins into biofuels. To achieve this, the mevalonate pathway was reconstructed into an E. coli chassis with six different terpene synthases (TSs). Strains containing the various TSs produced a spectrum of sesquiterpene compounds in minimal medium containing amino acids as the sole carbon source. The sesquiterpene production was optimized through three different regulation strategies using chamigrene synthase as an example. The highest total terpene titer reached 166 mg/L, and was achieved by applying a strategy to minimize mevalonate accumulation in vivo. The highest yields of total terpene were produced under reduced IPTG induction levels (0.25 mM), reduced induction temperature (25°C), and elevated substrate concentration (20 g/L amino acid mixture). A synthetic bioconversion consortium consisting of two engineering E. coli strains (DH1-TS and YH40-TS) with reconstructed terpene biosynthetic pathways was designed for comprehensive single-pot conversion of algal carbohydrates and proteins to

  20. Terpenes removal from biogas; Terpenenverwijdering uit biogas

    Energy Technology Data Exchange (ETDEWEB)

    Schulze, P.; Holstein, J.; De Haan, HR.; Vlap, H. [DNV KEMA, Arnhem (Netherlands)

    2013-06-15

    Biogas may contain unwanted and harmful components, including aromatic hydrocarbons such as terpenes. These terpenes (organic oils) are mainly present in citrus peel and plant residues; that is why especially raw biogas from organic waste digestion plants contains high concentrations of terpenes. If terpenes end up in the gas grid (with the injected biomethane) there is a risk that plastics (PE pipes) lose their mechanical properties by absorbing liquids or extracting ethereal plasticizers. This can lead to embrittlement greatly lowering the reliability of the piping. In addition, soft components are als o affected (gaskets and rubber O-rings). Besides the impact on the integrity of the gas grid, terpenes also mask the odor of natural gas odorants such as THT. This impedes the detection of gas leaks which is a significant security risk. Furthermore, the presence of terpenes in biogas leads to fouling of equipment used for the drying of biomethane, as well as contamination of adsorption liquids and membranes used in the upgrading process. Currently, terpenes are removed by activated carbon filters. The tool life of such a filter can be relatively short if terpene concentrations are high in the biogas; this results in a significant increase of the operational costs, due to the replacement of the carbon. This study looked at alternative techniques for removing much of the terpenes from biogas in a simple, efficient and cheap way. In a workshop with stakeholders two techniques were chosen to be tested on laboratory scale in order to demonstrate the proof of principle. These techniques are photo-oxydation and a gas scrubbing. Of all investigated techniques for the removal of limonene the application of UV radiation seems to be the most promising option because of the simplicity of the process, the high efficiency (up to 94%), the comparable operational costs with activated carbon (6.7 to 9.5 euro/kg limonene removed, compared to 10 euro/kg limonene removed for activated

  1. Terpene-induced porphyria and the illness of Vincent van Gogh

    Energy Technology Data Exchange (ETDEWEB)

    Lambrecht, R.; Cable, E.; Cable, J.; Clements, E.; Donohue, S.; Greene, Y.; Srivastava, K.; Arnold, W.; Bonkovsky, H. (Univ. of Massachusetts Medical Center, Worcester (United States) Univ. of Kansas Medical Center, Kansas City (United States))

    1992-01-01

    Vincent van Gogh suffered from recurrent bouts of an illness that may have been acute porphyria and abused camphor and alcohol, the latter particularly in the form of absinthe, a liqueur distilled from wormwood that was popular in 19th C France. To learn whether camphor or terpenes found in absinthe are porphyrogenic, the authors studied them in cultures of chick embryo liver cells. All were found to be porphyrogenic, especially in the presence of deferoxamine. The terpenes also induced the activity and protein amount of 5-aminolevulinate synthase and heme oxygenase, and induced activities of benzphetamine demethylase. The degree of porphyrin and enzyme induction produced by 1mM camphor was similar to that produced by 50uM glutethimide, a potent porphyrogen. Potency of pinene and thujone were lower. Camphor and glutethimide both produced accumulations of 8- and 7-COOH porphyrins, whereas pinene and thujone produced 4- and 2-COOH porphyrin accumulation. The authors conclude that camphor, pinen and thujone are porphyrogenic, cable of exacerbating acute porphyria, and may have done so in van Gogh.

  2. Selected oxidized fragrance terpenes are common contact allergens

    DEFF Research Database (Denmark)

    Matura, Mihaly; Sköld, Maria; Börje, Anna

    2005-01-01

    Terpenes are widely used fragrance compounds in fine fragrances, but also in domestic and occupational products. Terpenes oxidize easily due to autoxidation on air exposure. Previous studies have shown that limonene, linalool and caryophyllene are not allergenic themselves but readily form...... allergenic products on air-exposure. This study aimed to determine the frequency and characteristics of allergic reactions to selected oxidized fragrance terpenes other than limonene. In total 1511 consecutive dermatitis patients in 6 European dermatology centres were patch tested with oxidized fragrance...

  3. Reconstitution of a fungal meroterpenoid biosynthesis reveals the involvement of a novel family of terpene cyclases

    Science.gov (United States)

    Itoh, Takayuki; Tokunaga, Kinya; Matsuda, Yudai; Fujii, Isao; Abe, Ikuro; Ebizuka, Yutaka; Kushiro, Tetsuo

    2010-10-01

    Meroterpenoids are hybrid natural products of both terpenoid and polyketide origin. We identified a biosynthetic gene cluster that is responsible for the production of the meroterpenoid pyripyropene in the fungus Aspergillus fumigatus through reconstituted biosynthesis of up to five steps in a heterologous fungal expression system. The cluster revealed a previously unknown terpene cyclase with an unusual sequence and protein primary structure. The wide occurrence of this sequence in other meroterpenoid and indole-diterpene biosynthetic gene clusters indicates the involvement of these enzymes in the biosynthesis of various terpenoid-bearing metabolites produced by fungi and bacteria. In addition, a novel polyketide synthase that incorporated nicotinyl-CoA as the starter unit and a prenyltransferase, similar to that in ubiquinone biosynthesis, was found to be involved in the pyripyropene biosynthesis. The successful production of a pyripyropene analogue illustrates the catalytic versatility of these enzymes for the production of novel analogues with useful biological activities.

  4. Benzalacetone Synthase

    Directory of Open Access Journals (Sweden)

    Ikuro eAbe

    2012-03-01

    Full Text Available Benzalacetone synthase, from the medicinal plant Rheum palmatum (Polygonaceae (RpBAS, is a plant-specific chalcone synthase (CHS superfamily of type III polyketide synthase (PKS. RpBAS catalyzes the one-step, decarboxylative condensation of 4-coumaroyl-CoA with malonyl-CoA to produce the C6-C4 benzalacetone scaffold. The X-ray crystal structures of RpBAS confirmed that the diketide-forming activity is attributable to the characteristic substitution of the conserved active-site "gatekeeper" Phe with Leu. Furthermore, the crystal structures suggested that RpBAS employs novel catalytic machinery for the thioester bond cleavage of the enzyme-bound diketide intermediate and the final decarboxylation reaction to produce benzalacetone. Finally, by exploiting the remarkable substrate tolerance and catalytic versatility of RpBAS, precursor-directed biosynthesis efficiently generated chemically and structurally divergent, unnatural novel polyketide scaffolds. These findings provided a structural basis for the functional diversity of the type III PKS enzymes.

  5. Invertebrate Trehalose-6-Phosphate Synthase Gene: Genetic Architecture, Biochemistry, Physiological Function, and Potential Applications

    Directory of Open Access Journals (Sweden)

    Bin Tang

    2018-01-01

    Full Text Available The non-reducing disaccharide trehalose is widely distributed among various organisms. It plays a crucial role as an instant source of energy, being the major blood sugar in insects. In addition, it helps countering abiotic stresses. Trehalose synthesis in insects and other invertebrates is thought to occur via the trehalose-6-phosphate synthase (TPS and trehalose-6-phosphate phosphatase (TPP pathways. In many insects, the TPP gene has not been identified, whereas multiple TPS genes that encode proteins harboring TPS/OtsA and TPP/OtsB conserved domains have been found and cloned in the same species. The function of the TPS gene in insects and other invertebrates has not been reviewed in depth, and the available information is quite fragmented. The present review discusses the current understanding of the trehalose synthesis pathway, TPS genetic architecture, biochemistry, physiological function, and potential sensitivity to insecticides. We note the variability in the number of TPS genes in different invertebrate species, consider whether trehalose synthesis may rely only on the TPS gene, and discuss the results of in vitro TPS overexpression experiment. Tissue expression profile and developmental characteristics of the TPS gene indicate that it is important in energy production, growth and development, metamorphosis, stress recovery, chitin synthesis, insect flight, and other biological processes. We highlight the molecular and biochemical properties of insect TPS that make it a suitable target of potential pest control inhibitors. The application of trehalose synthesis inhibitors is a promising direction in insect pest control because vertebrates do not synthesize trehalose; therefore, TPS inhibitors would be relatively safe for humans and higher animals, making them ideal insecticidal agents without off-target effects.

  6. Structural determinants of reductive terpene cyclization in iridoid biosynthesis

    DEFF Research Database (Denmark)

    Kries, Hajo; Caputi, Lorenzo; Stevenson, Clare E M

    2016-01-01

    The carbon skeleton of ecologically and pharmacologically important iridoid monoterpenes is formed in a reductive cyclization reaction unrelated to canonical terpene cyclization. Here we report the crystal structure of the recently discovered iridoid cyclase (from Catharanthus roseus) bound...

  7. Selected oxidized fragrance terpenes are common contact allergens.

    Science.gov (United States)

    Matura, Mihaly; Sköld, Maria; Börje, Anna; Andersen, Klaus E; Bruze, Magnus; Frosch, Peter; Goossens, An; Johansen, Jeanne D; Svedman, Cecilia; White, Ian R; Karlberg, Ann-Therese

    2005-06-01

    Terpenes are widely used fragrance compounds in fine fragrances, but also in domestic and occupational products. Terpenes oxidize easily due to autoxidation on air exposure. Previous studies have shown that limonene, linalool and caryophyllene are not allergenic themselves but readily form allergenic products on air-exposure. This study aimed to determine the frequency and characteristics of allergic reactions to selected oxidized fragrance terpenes other than limonene. In total 1511 consecutive dermatitis patients in 6 European dermatology centres were patch tested with oxidized fragrance terpenes and some oxidation fractions and compounds. Oxidized linalool and its hydroperoxide fraction were found to be common contact allergens. Of the patients tested, 1.3% showed a positive reaction to oxidized linalool and 1.1% to the hydroperoxide fraction. About 0.5% of the patients reacted to oxidized caryophyllene whereas 1 patient reacted to oxidized myrcene. Of the patients reacting to the oxidized terpenes, 58% had fragrance-related contact allergy and/or a positive history for adverse reaction to fragrances. Autoxidation of fragrance terpenes contributes greatly to fragrance allergy, which emphasizes the need of testing with compounds that patients are actually exposed to and not only with the ingredients originally applied in commercial formulations.

  8. Transcriptome profiling of the Australian arid-land plant Eremophila serrulata (A.DC.) Druce (Scrophulariaceae) for the identification of monoterpene synthases.

    Science.gov (United States)

    Kracht, Octavia Natascha; Ammann, Ann-Christin; Stockmann, Julia; Wibberg, Daniel; Kalinowski, Jörn; Piotrowski, Markus; Kerr, Russell; Brück, Thomas; Kourist, Robert

    2017-04-01

    Plant terpenoids are a large and highly diverse class of metabolites with an important role in the immune defense. They find wide industrial application as active pharmaceutical ingredients, aroma and fragrance compounds. Several Eremophila sp. derived terpenoids have been documented. To elucidate the terpenoid metabolism, the transcriptome of juvenile and mature Eremophila serrulata (A.DC.) Druce (Scrophulariaceae) leaves was sequenced and a transcript library was generated. We report on the first transcriptomic dataset of an Eremophila plant. IlluminaMiSeq sequencing (2 × 300 bp) revealed 7,093,266 paired reads, which could be assembled to 34,505 isogroups. To enable detection of terpene biosynthetic genes, leaves were separately treated with methyl jasmonate, a well-documented inducer of plant secondary metabolites. In total, 21 putative terpene synthase genes were detected in the transcriptome data. Two terpene synthase isoenzymatic genes, termed ES01 and ES02, were successfully expressed in E. coli. The resulting proteins catalyzed the conversion of geranyl pyrophosphate, the universal substrate of monoterpene synthases to myrcene and Z-(b)-ocimene, respectively. The transcriptomic data and the discovery of the first terpene synthases from Eremophila serrulata are the initial step for the understanding of the terpene metabolism in this medicinally important plant genus. Copyright © 2017 Elsevier Ltd. All rights reserved.

  9. Seasonal influence on gene expression of monoterpene synthases in Salvia officinalis (Lamiaceae).

    Science.gov (United States)

    Grausgruber-Gröger, Sabine; Schmiderer, Corinna; Steinborn, Ralf; Novak, Johannes

    2012-03-01

    Garden sage (Salvia officinalis L., Lamiaceae) is one of the most important medicinal and aromatic plants and possesses antioxidant, antimicrobial, spasmolytic, astringent, antihidrotic and specific sensorial properties. The essential oil of the plant, formed mainly in very young leaves, is in part responsible for these activities. It is mainly composed of the monoterpenes 1,8-cineole, α- and β-thujone and camphor synthesized by the 1,8-cineole synthase, the (+)-sabinene synthase and the (+)-bornyl diphosphate synthase, respectively, and is produced and stored in epidermal glands. In this study, the seasonal influence on the formation of the main monoterpenes in young, still expanding leaves of field-grown sage plants was studied in two cultivars at the level of mRNA expression, analyzed by qRT-PCR, and at the level of end-products, analyzed by gas chromatography. All monoterpene synthases and monoterpenes were significantly influenced by cultivar and season. 1,8-Cineole synthase and its end product 1,8-cineole remained constant until August and then decreased slightly. The thujones increased steadily during the vegetative period. The transcript level of their corresponding terpene synthase, however, showed its maximum in the middle of the vegetative period and declined afterwards. Camphor remained constant until August and then declined, exactly correlated with the mRNA level of the corresponding terpene synthase. In summary, terpene synthase mRNA expression and respective end product levels were concordant in the case of 1,8-cineole (r=0.51 and 0.67 for the two cultivars, respectively; p<0.05) and camphor (r=0.75 and 0.82; p<0.05) indicating basically transcriptional control, but discordant for α-/β-thujone (r=-0.05 and 0.42; p=0.87 and 0.13, respectively). Copyright © 2011 Elsevier GmbH. All rights reserved.

  10. Downregulation of dTps1 in Drosophila melanogaster larvae confirms involvement of trehalose in redox regulation following desiccation.

    Science.gov (United States)

    Thorat, Leena; Mani, Krishna-Priya; Thangaraj, Pradeep; Chatterjee, Suvro; Nath, Bimalendu B

    2016-03-01

    As a survival strategy to environmental water deficits, desiccation-tolerant organisms are commonly known for their ability to recruit stress-protective biomolecules such as trehalose. We have previously reported the pivotal role of trehalose in larval desiccation tolerance in Drosophila melanogaster. Trehalose has emerged as a versatile molecule, serving mainly as energy source in insects and also being a stress protectant. While several recent reports have revealed the unconventional role of trehalose in scavenging reactive oxygen species in yeast and plants, this aspect has not received much attention in animals. We examined the status of desiccation-induced generation of reactive oxygen species in D. melanogaster larvae and the possible involvement of trehalose in ameliorating the harmful consequences thereof. Insect trehalose synthesis is governed by the enzyme trehalose 6-phosphate synthase 1 (TPS1). Using the ubiquitous da-GAL4-driven expression of the dTps1-RNAi transgene, we generated dTps1-downregulated Drosophila larvae possessing depleted levels of dTps1 transcripts. This resulted in the inability of the larvae for trehalose synthesis, thereby allowing us to elucidate the significance of trehalose in the regulation of desiccation-responsive redox homeostasis. Furthermore, the results from molecular genetics studies, biochemical assays, electron spin resonance analyses and a simple, non-invasive method of whole larval live imaging suggested that trehalose in collaboration with superoxide dismutase (SOD) is involved in the maintenance of redox state in D. melanogaster.

  11. Antimalarial activity of the terpene nerolidol.

    Science.gov (United States)

    Saito, Alexandre Y; Marin Rodriguez, Adriana A; Menchaca Vega, Danielle S; Sussmann, Rodrigo A C; Kimura, Emília A; Katzin, Alejandro M

    2016-12-01

    Malaria, an infectious disease that kills more than 438,000 people per year worldwide, is a major public health problem. The emergence of strains resistant to conventional therapeutic agents necessitates the discovery of new drugs. We previously demonstrated that various substances, including terpenes, have antimalarial activity in vitro and in vivo. Nerolidol is a sesquiterpene present as an essential oil in several plants that is used in scented products and has been approved by the US Food and Drug Administration as a food-flavouring agent. In this study, the antimalarial activity of nerolidol was investigated in a mouse model of malaria. Mice were infected with Plasmodium berghei ANKA and were treated with 1000 mg/kg/dose nerolidol in two doses delivered by the oral or inhalation route. In mice treated with nerolidol, parasitaemia was inhibited by >99% (oral) and >80% (inhalation) until 14 days after infection (P  0.05). The toxicity of nerolidol administered by either route was not significant, whilst genotoxicity was observed only at the highest dose tested. These results indicate that combined use of nerolidol and other drugs targeting different points of the same isoprenoid pathway may be an effective treatment for malaria. Copyright © 2016 Elsevier B.V. and International Society of Chemotherapy. All rights reserved.

  12. Terpene and dextran renewable resources for the synthesis of amphiphilic biopolymers.

    Science.gov (United States)

    Alvès, Marie-Hélène; Sfeir, Huda; Tranchant, Jean-François; Gombart, Emilie; Sagorin, Gilles; Caillol, Sylvain; Billon, Laurent; Save, Maud

    2014-01-13

    The present work shows the synthesis of amphiphilic polymers based on the hydrophilic dextran and the hydrophobic terpenes as renewable resources. The first step concerns the synthesis of functional terpene molecules by thiol-ene addition chemistry involving amino or carboxylic acid thiols and dihydromyrcenol terpene. The terpene-modified polysaccharides were subsequently synthesized by coupling the functional terpenes with dextran. A reductive amination step produced terpene end-modified dextran with 94% of functionalization, while the esterification step produced three terpene-grafted dextrans with a number of terpene units per dextran of 1, 5, and 10. The amphiphilic renewable grafted polymers were tested as emulsifiers for the stabilization of liquid miniemulsion of terpene droplets dispersed in an aqueous phase. The average hydrodynamic diameter of the stable droplets was observed at about 330 nm.

  13. Structural and In Vivo Studies on Trehalose-6-Phosphate Synthase from Pathogenic Fungi Provide Insights into Its Catalytic Mechanism, Biological Necessity, and Potential for Novel Antifungal Drug Design

    Directory of Open Access Journals (Sweden)

    Yi Miao

    2017-07-01

    Full Text Available The disaccharide trehalose is critical to the survival of pathogenic fungi in their human host. Trehalose-6-phosphate synthase (Tps1 catalyzes the first step of trehalose biosynthesis in fungi. Here, we report the first structures of eukaryotic Tps1s in complex with substrates or substrate analogues. The overall structures of Tps1 from Candida albicans and Aspergillus fumigatus are essentially identical and reveal N- and C-terminal Rossmann fold domains that form the glucose-6-phosphate and UDP-glucose substrate binding sites, respectively. These Tps1 structures with substrates or substrate analogues reveal key residues involved in recognition and catalysis. Disruption of these key residues severely impaired Tps1 enzymatic activity. Subsequent cellular analyses also highlight the enzymatic function of Tps1 in thermotolerance, yeast-hypha transition, and biofilm development. These results suggest that Tps1 enzymatic functionality is essential for the fungal stress response and virulence. Furthermore, structures of Tps1 in complex with the nonhydrolyzable inhibitor, validoxylamine A, visualize the transition state and support an internal return-like catalytic mechanism that is generalizable to other GT-B-fold retaining glycosyltransferases. Collectively, our results depict key Tps1-substrate interactions, unveil the enzymatic mechanism of these fungal proteins, and pave the way for high-throughput inhibitor screening buttressed and guided by the current structures and those of high-affinity ligand-Tps1 complexes.

  14. Progress in terpene synthesis strategies through engineering of Saccharomyces cerevisiae.

    Science.gov (United States)

    Paramasivan, Kalaivani; Mutturi, Sarma

    2017-12-01

    Terpenes are natural products with a remarkable diversity in their chemical structures and they hold a significant market share commercially owing to their distinct applications. These potential molecules are usually derived from terrestrial plants, marine and microbial sources. In vitro production of terpenes using plant tissue culture and plant metabolic engineering, although receiving some success, the complexity in downstream processing because of the interference of phenolics and product commercialization due to regulations that are significant concerns. Industrial workhorses' viz., Escherichia coli and Saccharomyces cerevisiae have become microorganisms to produce non-native terpenes in order to address critical issues such as demand-supply imbalance, sustainability and commercial viability. S. cerevisiae enjoys several advantages for synthesizing non-native terpenes with the most significant being the compatibility for expressing cytochrome P450 enzymes from plant origin. Moreover, achievement of high titers such as 40 g/l of amorphadiene, a sesquiterpene, boosts commercial interest and encourages the researchers to envisage both molecular and process strategies for developing yeast cell factories to produce these compounds. This review contains a brief consideration of existing strategies to engineer S. cerevisiae toward the synthesis of terpene molecules. Some of the common targets for synthesis of terpenes in S. cerevisiae are as follows: overexpression of tHMG1, ERG20, upc2-1 in case of all classes of terpenes; repression of ERG9 by replacement of the native promoter with a repressive methionine promoter in case of mono-, di- and sesquiterpenes; overexpression of BTS1 in case of di- and tetraterpenes. Site-directed mutagenesis such as Upc2p (G888A) in case of all classes of terpenes, ERG20p (K197G) in case of monoterpenes, HMG2p (K6R) in case of mono-, di- and sesquiterpenes could be some generic targets. Efforts are made to consolidate various studies

  15. Production of terpenes in the culture of Chlorophyceae and Rhodophyta

    Science.gov (United States)

    Abe, M.; Hashimoto, S.

    2014-12-01

    Terpenes show high reactivity in the troposphere, contributing to organic aerosol reactions with OH radicals. One of the main sources of terpenes in the atmosphere is terrestrial plants. It has been recently reported that marine phytoplankton also produce monoterpenes (Yassaa et al: 2008). Because aerosol production of natural origin affects the cloud cover over the open ocean, it is important to investigate the origin of aerosol generation in the open ocean. In this study, we investigated the production of terpenes and isoprene with a focus on Chlamydomonas (Chlorophyceae) and Rhodella maculata (Rhodophyta). Concentrations of terpenes and isoprene were measured using a dynamic headspace (GERSTEL DHS)—gas chromatograph (Agilent 6890N)—mass spectrometer (Agilent 5975C). In addition, chlorophyll a was measured using a fluorometer (Turner TD-700). The results showed that isoprene, α-pinene, and β-pinene were produced by Chlamydomonas sp. and that isoprene, limonene, and camphene were produced by Rhodella maculata. Chlamydomonas sp. produced α-pinene and β-pinene, similar to land plants. The ratio of the pinene/isoprene concentrations in the atmosphere over seawater where phytoplankton are blooming has been reported as approximately 0.7 (Yassaa et al: 2008). In this experiment, the pinene/isoprene concentration ratios in the cultures were approximately 0.1. This result indicates that marine phytoplankton may not be ignored in the marine atmosphere chemistry of terpenes.

  16. Mechanism of Action and Inhibition of dehydrosqualene Synthase

    Energy Technology Data Exchange (ETDEWEB)

    F Lin; C Liu; Y Liu; Y Zhang; K Wang; W Jeng; T Ko; R Cao; A Wang; E Oldfield

    2011-12-31

    'Head-to-head' terpene synthases catalyze the first committed steps in sterol and carotenoid biosynthesis: the condensation of two isoprenoid diphosphates to form cyclopropylcarbinyl diphosphates, followed by ring opening. Here, we report the structures of Staphylococcus aureus dehydrosqualene synthase (CrtM) complexed with its reaction intermediate, presqualene diphosphate (PSPP), the dehydrosqualene (DHS) product, as well as a series of inhibitors. The results indicate that, on initial diphosphate loss, the primary carbocation so formed bends down into the interior of the protein to react with C2,3 double bond in the prenyl acceptor to form PSPP, with the lower two-thirds of both PSPP chains occupying essentially the same positions as found in the two farnesyl chains in the substrates. The second-half reaction is then initiated by the PSPP diphosphate returning back to the Mg{sup 2+} cluster for ionization, with the resultant DHS so formed being trapped in a surface pocket. This mechanism is supported by the observation that cationic inhibitors (of interest as antiinfectives) bind with their positive charge located in the same region as the cyclopropyl carbinyl group; that S-thiolo-diphosphates only inhibit when in the allylic site; activity results on 11 mutants show that both DXXXD conserved domains are essential for PSPP ionization; and the observation that head-to-tail isoprenoid synthases as well as terpene cyclases have ionization and alkene-donor sites which spatially overlap those found in CrtM.

  17. Menthol differs from other terpenic essential oil constituents.

    Science.gov (United States)

    Kolassa, Norbert

    2013-02-01

    The European Medicines Agency concluded that there is a risk of suppositories containing terpenic derivatives, which are used to treat coughs and colds, inducing neurological disorders, especially convulsions, in infants and small children. Terpenic derivatives are found in essential oils obtained from plants and include camphor, eucalyptol (syn. 1,8-cineol), thujone, and menthol. Chemistry, pharmacodynamics and pharmacokinetics of these compounds are clearly different and explain the appearance of convulsions following camphor, thujone, and eucalyptus oil overdose/poisoning, whereas no convulsions have been reported in cases of menthol overdose/poisoning in accordance with the pharmacological properties of menthol. Thus, a general verdict on all terpenic derivatives without differentiation appears inappropriate. Copyright © 2012 Elsevier Inc. All rights reserved.

  18. Converting S-limonene synthase to pinene or phellandrene synthases reveals the plasticity of the active site.

    Science.gov (United States)

    Xu, Jinkun; Ai, Ying; Wang, Jianhui; Xu, Jingwei; Zhang, Yongkang; Yang, Dong

    2017-05-01

    S-limonene synthase is a model monoterpene synthase that cyclizes geranyl pyrophosphate (GPP) to form S-limonene. It is a relatively specific enzyme as the majority of its products are composed of limonene. In this study, we converted it to pinene or phellandrene synthases after introducing N345A/L423A/S454A or N345I mutations. Further studies on N345 suggest the polarity of this residue plays a critical role in limonene production by stabilizing the terpinyl cation intermediate. If it is mutated to a non-polar residue, further cyclization or hydride shifts occurs so the carbocation migrates towards the pyrophosphate, leading to the production of pinene or phellandrene. On the other hand, mutant enzymes that still possess a polar residue at this position produce limonene as the major product. N345 is not the only polar residue that may stabilize the terpinyl cation because it is not strictly conserved among limonene synthases across species and there are also several other polar residues in this area. These residues could form a "polar pocket" that may collectively play this stabilizing role. Our study provides important insights into the catalytic mechanism of limonene synthases. Furthermore, it also has wider implications on the evolution of terpene synthases. Copyright © 2017 Elsevier Ltd. All rights reserved.

  19. Oil production at different stages of leaf development in Lippia alba

    OpenAIRE

    Diego Pandeló; Talita D. Melo; Júnya L. Singulani; Fernanda A. F. Guedes; Marco A. Machado; Cíntia M. Coelho; Lyderson F. Viccini; Marcelo O. Santos

    2012-01-01

    The aim of this work was to analyze terpene oil production and terpene synthases (TPS) gene expression from leaves at different developmental stages of different chemotypes of Lippia alba (Mill.) N.E. Br. ex Britton & P. Wilson, Verbenaceae. Hydro-distilled essential oil were used for chemical analysis and gene expression of three monoterpene synthase genes called LaTPS12, LaTPS23 and LaTPS25 were used for analyses of gene expression associated to oil production. The putative genes were a...

  20. Sesquiterpene Synthase-3-Hydroxy-3-Methylglutaryl Coenzyme A Synthase Fusion Protein Responsible for Hirsutene Biosynthesis in Stereum hirsutum.

    Science.gov (United States)

    Flynn, Christopher M; Schmidt-Dannert, Claudia

    2018-06-01

    The wood-rotting mushroom Stereum hirsutum is a known producer of a large number of namesake hirsutenoids, many with important bioactivities. Hirsutenoids form a structurally diverse and distinct class of sesquiterpenoids. No genes involved in hirsutenoid biosynthesis have yet been identified or their enzymes characterized. Here, we describe the cloning and functional characterization of a hirsutene synthase as an unexpected fusion protein of a sesquiterpene synthase (STS) with a C-terminal 3-hydroxy-3-methylglutaryl-coenzyme A (3-hydroxy-3-methylglutaryl-CoA) synthase (HMGS) domain. Both the full-length fusion protein and truncated STS domain are highly product-specific 1,11-cyclizing STS enzymes with kinetic properties typical of STSs. Complementation studies in Saccharomyces cerevisiae confirmed that the HMGS domain is also functional in vivo Phylogenetic analysis shows that the hirsutene synthase domain does not form a clade with other previously characterized sesquiterpene synthases from Basidiomycota. Comparative gene structure analysis of this hirsutene synthase with characterized fungal enzymes reveals a significantly higher intron density, suggesting that this enzyme may be acquired by horizontal gene transfer. In contrast, the HMGS domain is clearly related to other fungal homologs. This STS-HMGS fusion protein is part of a biosynthetic gene cluster that includes P450s and oxidases that are expressed and could be cloned from cDNA. Finally, this unusual fusion of a terpene synthase to an HMGS domain, which is not generally recognized as a key regulatory enzyme of the mevalonate isoprenoid precursor pathway, led to the identification of additional HMGS duplications in many fungal genomes, including the localization of HMGSs in other predicted sesquiterpenoid biosynthetic gene clusters. IMPORTANCE Hirsutenoids represent a structurally diverse class of bioactive sesquiterpenoids isolated from fungi. Identification of their biosynthetic pathways will provide

  1. Biotransformation of geosmin by terpene-degrading bacteria.

    Science.gov (United States)

    Two terpene-degrading bacteria that are able to transform geosmin have been identified. Pseudomonas sp. SBR3-tpnb, isolated on -terpinene, converts geosmin to several products; the major products are keto-geosmins. This geosmin transformation ability is inducible by -terpinene. Rhodococcus wratisl...

  2. Transfer of Orally Administered Terpenes in Goat Milk and Cheese

    Directory of Open Access Journals (Sweden)

    I. Poulopoulou

    2012-10-01

    Full Text Available The objective of the present study was to investigate the relationships between terpenes’ intake and their presence in animal tissues (blood and milk as well as in the final product (cheese. Eight dairy goats were divided in two balanced groups, representing control (C and treatment (T group. In T group oral administration of a mixture of terpenes (α-pinene, limonene and β-caryophyllene was applied over a period of 18 d. Cheese was produced, from C and T groups separately, on three time points, twice during the period of terpenes’ oral administration and once after the end of experiment. Terpenes were identified in blood by extraction using petroleum ether and in milk and cheese by the use of solid phase micro-extraction (SPME method, followed by GC-MS analysis. Chemical properties of the milk and the produced cheeses were analyzed and found not differing between the two groups. Limonene and α-pinene were found in all blood and milk samples of the T group after a lag-phase of 3 d, while β-caryophyllene was determined only in few milk samples. Moreover, none of the terpenes were traced in blood and milk of C animals. In cheese, terpenes’ concentrations presented a more complicated pattern implying that terpenes may not be reliable feed tracers. We concluded that monoterpenes can be regarded as potential feed tracers for authentification of goat milk, but further research is required on factors affecting their transfer.

  3. An (E,E)-a-farnesene synthase gene of soybean has a role in defense against nematodes and is involved in synthesizing insect-induced volatiles

    Science.gov (United States)

    Plant terpene synthase genes (TPSs) have roles in diverse biological processes. Here we report the functional characterization of one member of the soybean TP S gene family, which was designated GmAFS. Recombinant GmAFS produced in E.coli catalyzed the formation of a sesquiterpene (E,E)-a-farnesene....

  4. Oxidatively Stable Flexible Aerogel Composites for Reusable TPS, Phase I

    Data.gov (United States)

    National Aeronautics and Space Administration — NASA?s Next Generation Launch Vehicle Technology Program has an interest in robust TPS materials with the highest level of thermal performance at the lowest possible...

  5. Fiber-Coupled Spectrometer for TPS Materials, Phase I

    Data.gov (United States)

    National Aeronautics and Space Administration — Heat shield technology is a critical component of manned spaceflight. In particular, the new Crew Exploration Vehicle (CEV) requires thermal protection systems (TPS)...

  6. Monoterpene synthase from Dracocephalum kotschyi and SPME-GC-MS analysis of its aroma profile

    Directory of Open Access Journals (Sweden)

    S. Saeidnia

    2014-04-01

    Full Text Available Dracocephalum kotschyi (Lamiaceae, as one of the remarkable aromatic plants, widely grows and also is cultivated in various temperate regions of Iran. There are diverse reports about the composition of the oil of this plant representing limonene derivatives as its major compounds. There is no report on cloning of mono- or sesquiterpene synthases from this plant. In the present study, the aroma profile of D. kotschyi has been extracted and analyzed via Headspace Solid-Phase Microextraction technique coupled with Gas Chromatography- Mass Spectroscopy. In order to determine the sequence of the active terpene synthase in this plant, first mRNA was prepared and cloning was performed by 3’ and 5’-RACEs-PCR method, then cDNA was sequenced and finally aligned with other recognized terpene synthases. The results showed that the plant leaves mainly comprised geranial (37.2%, limonene-10-al (28.5%, limonene (20.1% and 1,1-dimethoxy decane (14.5%. Sequencing the cDNA cloned from this plant revealed the presence of a monoterpene synthase absolutely similar to limonene synthase, responsible in formation of limonene, terpinolene, camphene and some other cyclic monoterpenes in its young leaves.

  7. Jejak Karbon Pengolahan Sampah Di Tps Tlogomas Malang

    OpenAIRE

    Sunarto; P. Hadi, Sudharto; Purwanto

    2014-01-01

    JEJAK KARBON PENGOLAHAN SAMPAH DI tps tlogomas malang Carbon Footprint of Solid Waste Processing At TPS Tlogomas MalangSunarto1, Sudharto P. Hadi2, Purwanto31,2,3Program Doktor Ilmu Lingkungan Universitas DiponegoroAlamat korespondensi : Jl. Imam Bardjo, SH No. 3 Semarang 50241Email: 1) , 2) ste sector is one of human activities that cause global warming. Decomposition of organic waste in landfill produces greenhouse gas emissions in the form...

  8. Antibacterial activity of sulfamethoxazole transformation products (TPs): general relevance for sulfonamide TPs modified at the para position.

    Science.gov (United States)

    Majewsky, Marius; Wagner, Danny; Delay, Markus; Bräse, Stefan; Yargeau, Viviane; Horn, Harald

    2014-10-20

    Sulfonamide antibiotics undergo transformation in the aquatic environment through biodegradation, photolysis, or hydrolysis. In this study, the residual antibacterial activity of 11 transformation products (TPs) of sulfamethoxazole (SMX) was investigated with regard to their in vitro growth and luminescence inhibition on Vibrio fischeri (30 min and 24 h exposure). Two transformation products, 4-hydroxy-SMX and N(4)-hydroxy-acetyl-SMX, were synthesized in-house and confirmed by nuclear magnetic resonance and high-resolution mass spectrometry. Results of individual compound experiments showed that TPs modified at the para amino group still exhibit clear antibacterial effects, whereas TPs resulting from breakdown of the SMX structure lost this mechanism of action. 4-NO2- and 4-OH-SMX were found to inhibit growth to a clearly greater extent than the parent compound, SMX. In contrast, the N(4)-acetyl- and N(4)-hydroxy-acetyl-derivatives retain less than 10 and 5% of the effect of SMX on growth and luminescence inhibition, respectively. The effect of a mixture of para-modified TPs was observed to be additive. Considering the homologous series of sulfa drugs widely prescribed and their common mechanism of action, the potential environmental impact must consider the total amount of sulfonamide antibiotics and their derivative TPs, which might end up in a water body. Extrapolating the results obtained here for the para TPs of SMX to other sulfa drugs and determining the persistence and occurrence of these compounds in the aquatic environment is required for improved risk assessment.

  9. Role of TPS in 125I brachytherapy for orbital tumors

    International Nuclear Information System (INIS)

    Ren Ling; Dai Haojie; Li Quan

    2012-01-01

    Objective: To investigate the role of TPS in 125 I brachytherapy for orbital tumors. Methods: Sixty-six patients with orbital tumor treated with 125 I seeds from 2005 to 2009 were retrospectively analyzed. Forty-three patients were treated using TPS guided brachytherapy and the prescribed dose was 140 Gy. Other 23 patients were treated without TPS but simply implanted with 125 I seeds at 1 cm intervals in parallel with each other intraoperatively. CT and TPS quality verification were performed postoperatively in all patients. Also, CT and (or) MRI examination were performed at 3, 6, 12 and 24 months after brachytherapy for follow-up. χ 2 test and Kaplan-Meier survival analysis with log-rank significance test were used with SPSS 17.0. Results: A total of 1070 125 I seeds were implanted in 66 cases, on average, (16.2 ± 7.3) seeds for each patient. The satisfaction rates of postoperative quality verification in patients with and without TPS pre-plans were 79.07% (34/43) and 43.48% (10/23) respectively (χ 2 =8.542, P=0.003). Ten patients were lost in follow-up. Local recurrence rates in patients with favorable postoperative quality verification were 0 (0/37) in 3 months, 6.25% (2/32) in 6 months, 13.64% (3/22) in 12 months and 3/9 in 24 months respectively, which were significantly different from those (5.26% (1/19), 16.67% (3/18), 30.77% (4/13), 6/6) in the patients with inferior postoperative quality verification (χ 2 =9.017, P=0.0003). Conclusions: TPS plays an important role in 125 I brachytherapy for orbital tumors. Also, postoperative quality verification by TPS may help predict the local recurrence after brachytherapy. (authors)

  10. Catalytic Synthesis and Antifungal Activity of New Polychlorinated Natural Terpenes

    Directory of Open Access Journals (Sweden)

    Hana Ighachane

    2017-01-01

    Full Text Available Various unsaturated natural terpenes were selectively converted to the corresponding polychlorinated products in good yields using iron acetylacetonate in combination with nucleophilic cocatalyst. The synthesized compounds were evaluated for their in vitro antifungal activity. The antifungal bioassays showed that 2c and 2d possessed significant antifungal activity against Fusarium oxysporum f. sp. albedinis (Foa, Fusarium oxysporum f. sp. canariensis (Foc, and Verticillium dahliae (Vd.

  11. TPS/LDPE blends reinforced with lignocellulose fibers; Compositos TPS/LDPE reforcados com fibras lignocelulosicas

    Energy Technology Data Exchange (ETDEWEB)

    Lopes, G.K.; Andrade, C.T., E-mail: kloc@ima.ufrj.b [Universidade Federal do Rio de Janeiro (IMA/UFRJ), RJ (Brazil). Inst. de Macromoleculas Eloisa Mano

    2010-07-01

    Because of their abundance, availability, low abrasiveness and mechanical properties, cellulose fibers have been frequently chosen as reinforcing fillers in composites. Castor bean cake, the residue from biodiesel production, is rich in lignocellulose fibers and proteins. One of these proteins is ricin, a toxin protein. In this work, ricin was denatured by heat treatment in water at 90 deg C for 4 h. Thermoplastic starch (TPS), low density polyethylene (LDPE), maleated polyethylene (used as the compatibilizing agent), and an organophilic clay were processed in the presence of different contents of heat treated castor bean cake. Processing was carried out in a single-screw extruder, at 400 rpm, with heat zones at 130 deg C, 135 deg C, 135 deg C and 130 deg C (from feed zone to die end). The structural and mechanical properties of the resulting polymeric composites were investigated, and revealed the reinforcing effect of the partially purified cellulose fibers. (author)

  12. Induced terpene accumulation in Norway spruce inhibits bark beetle colonization in a dose-dependent manner.

    Directory of Open Access Journals (Sweden)

    Tao Zhao

    Full Text Available Tree-killing bark beetles (Coleoptera, Scolytinae are among the most economically and ecologically important forest pests in the northern hemisphere. Induction of terpenoid-based oleoresin has long been considered important in conifer defense against bark beetles, but it has been difficult to demonstrate a direct correlation between terpene levels and resistance to bark beetle colonization.To test for inhibitory effects of induced terpenes on colonization by the spruce bark beetle Ips typographus (L. we inoculated 20 mature Norway spruce Picea abies (L. Karsten trees with a virulent fungus associated with the beetle, Ceratocystis polonica (Siem. C. Moreau, and investigated induced terpene levels and beetle colonization in the bark.Fungal inoculation induced very strong and highly variable terpene accumulation 35 days after inoculation. Trees with high induced terpene levels (n = 7 had only 4.9% as many beetle attacks (5.1 vs. 103.5 attacks m(-2 and 2.6% as much gallery length (0.029 m m(-2 vs. 1.11 m m(-2 as trees with low terpene levels (n = 6. There was a highly significant rank correlation between terpene levels at day 35 and beetle colonization in individual trees. The relationship between induced terpene levels and beetle colonization was not linear but thresholded: above a low threshold concentration of ∼100 mg terpene g(-1 dry phloem trees suffered only moderate beetle colonization, and above a high threshold of ∼200 mg terpene g(-1 dry phloem trees were virtually unattacked.This is the first study demonstrating a dose-dependent relationship between induced terpenes and tree resistance to bark beetle colonization under field conditions, indicating that terpene induction may be instrumental in tree resistance. This knowledge could be useful for developing management strategies that decrease the impact of tree-killing bark beetles.

  13. Woven TPS Enabling Missions Beyond Heritage Carbon Phenolic

    Science.gov (United States)

    Stackpoole, Margaret M.; Venkatapathy, Ethiraj; Feldman, Jay D.

    2013-01-01

    NASAs Office of the Chief Technologist (OCT) Game Changing Division recently funded an effort to advance a Woven TPS (WTPS) concept. WTPS is a new approach to producing TPS architectures that uses precisely engineered 3D weaving techniques to customize material characteristics needed to meet specific missions requirements for protecting space vehicles from the intense heating generated during atmospheric entry. Using WTPS, sustainable, scalable, mission-optimized TPS solutions can be achieved with relatively low life cycle costs compared with the high costs and long development schedules currently associated with material development and certification. WTPS leverages the mature state-of-the-art weaving technology that has evolved from the textile industry to design TPS materials with tailorable performance. Currently, missions anticipated encountering heat fluxes in the range of 1500 4000 Wcm2 and pressures greater than 1.5 atm are limited to using fully dense Carbon Phenolic. However, fully dense carbon phenolic is only mass efficient at higher heat fluxes g(reater than 4000 Wcm2), and current mission designs suffer this mass inefficiency for lack of an alternative mid-density TPS. WTPS not only bridges this mid-density TPS gap but also offers a replacement for carbon phenolic, which itself requires a significant and costly redevelopment effort to re-establish its capability for use in the high heat flux missions recently prioritized in the NRC Decadal survey, including probe missions to Venus, Saturn and Neptune. This presentation will overview the WTPS concept and present some results from initial testing completed comparing WTPS architectures to heritage carbon phenolic.

  14. Actividad antimicobacteriana de terpenos Antimycobacterial activity of terpenes

    Directory of Open Access Journals (Sweden)

    Juan Gabriel Bueno-Sánchez

    2009-12-01

    Full Text Available Introducción: La tuberculosis (TB, causada por Mycobacterium tuberculosis es la mayor causa de mortalidad mundial por un único agente patógeno. Asimismo, un gran número de micobacterias no tuberculosas, especialmente M. avium, M. intracellulare y M. chelonae, causan infecciones oportunistas en pacientes con SIDA. Muchos terpenos poseen actividad biológica y se emplean en el tratamiento de diversas enfermedades, razón que los hace fuente de moléculas promisorias. Objetivo: El objetivo del presente estudio fue determinar la actividad antimicobacteriana de 16 terpenos contra M. tuberculosis H37Rv y un aislamiento clínico de M. chelonae. Materiales y métodos: Se obtuvo la concentración mínima inhibitoria (CMI de los mismos y se realizaron curvas de letalidad para establecer actividad bactericida, empleando una técnica de macrodilución en caldo estandarizada para este tipo de compuestos volátiles. Resultados: Los terpenos con menor valor de CMI fueron timol y carvacrol, con concentraciones de 125-250 μg/mL, y actividad bactericida contra los dos microorganismos. Geraniol, mirceno, ρ-cimeno, alfa-pineno, presentaron valores de CMI entre 250 y 500 μg/mL. Conclusiones: Algunos terpenos han presentado actividad importante contra microorganismos del género Mycobacterium, sin embargo los valores de CMI obtenidos no explican el efecto antimicrobiano presentado por el aceite completo, se requiere evaluar las interacciones de sinergismo y/o antagonismo entre los terpenos para determinar los componentes responsables de la acción farmacológica. Salud UIS 2009; 41: 231-235Introduction: Tuberculosis (TB caused by Mycobacterium tuberculosis is the major source of global mortality from a single pathogen. Moreover, a large number of nontuberculous mycobacteria, especially M. avium, M. intracellulare and M. chelonae, cause opportunistic infection in AIDS patients. Terpenes, possess a wide spectrum of biological activity and are used in the

  15. Evolution of Conifer Diterpene Synthases: Diterpene Resin Acid Biosynthesis in Lodgepole Pine and Jack Pine Involves Monofunctional and Bifunctional Diterpene Synthases1[W][OA

    Science.gov (United States)

    Hall, Dawn E.; Zerbe, Philipp; Jancsik, Sharon; Quesada, Alfonso Lara; Dullat, Harpreet; Madilao, Lina L.; Yuen, Macaire; Bohlmann, Jörg

    2013-01-01

    Diterpene resin acids (DRAs) are major components of pine (Pinus spp.) oleoresin. They play critical roles in conifer defense against insects and pathogens and as a renewable resource for industrial bioproducts. The core structures of DRAs are formed in secondary (i.e. specialized) metabolism via cycloisomerization of geranylgeranyl diphosphate (GGPP) by diterpene synthases (diTPSs). Previously described gymnosperm diTPSs of DRA biosynthesis are bifunctional enzymes that catalyze the initial bicyclization of GGPP followed by rearrangement of a (+)-copalyl diphosphate intermediate at two discrete class II and class I active sites. In contrast, similar diterpenes of gibberellin primary (i.e. general) metabolism are produced by the consecutive activity of two monofunctional class II and class I diTPSs. Using high-throughput transcriptome sequencing, we discovered 11 diTPS from jack pine (Pinus banksiana) and lodgepole pine (Pinus contorta). Three of these were orthologous to known conifer bifunctional levopimaradiene/abietadiene synthases. Surprisingly, two sets of orthologous PbdiTPSs and PcdiTPSs were monofunctional class I enzymes that lacked functional class II active sites and converted (+)-copalyl diphosphate, but not GGPP, into isopimaradiene and pimaradiene as major products. Diterpene profiles and transcriptome sequences of lodgepole pine and jack pine are consistent with roles for these diTPSs in DRA biosynthesis. The monofunctional class I diTPSs of DRA biosynthesis form a new clade within the gymnosperm-specific TPS-d3 subfamily that evolved from bifunctional diTPS rather than monofunctional enzymes (TPS-c and TPS-e) of gibberellin metabolism. Homology modeling suggested alterations in the class I active site that may have contributed to their functional specialization relative to other conifer diTPSs. PMID:23370714

  16. Evolution of conifer diterpene synthases: diterpene resin acid biosynthesis in lodgepole pine and jack pine involves monofunctional and bifunctional diterpene synthases.

    Science.gov (United States)

    Hall, Dawn E; Zerbe, Philipp; Jancsik, Sharon; Quesada, Alfonso Lara; Dullat, Harpreet; Madilao, Lina L; Yuen, Macaire; Bohlmann, Jörg

    2013-02-01

    Diterpene resin acids (DRAs) are major components of pine (Pinus spp.) oleoresin. They play critical roles in conifer defense against insects and pathogens and as a renewable resource for industrial bioproducts. The core structures of DRAs are formed in secondary (i.e. specialized) metabolism via cycloisomerization of geranylgeranyl diphosphate (GGPP) by diterpene synthases (diTPSs). Previously described gymnosperm diTPSs of DRA biosynthesis are bifunctional enzymes that catalyze the initial bicyclization of GGPP followed by rearrangement of a (+)-copalyl diphosphate intermediate at two discrete class II and class I active sites. In contrast, similar diterpenes of gibberellin primary (i.e. general) metabolism are produced by the consecutive activity of two monofunctional class II and class I diTPSs. Using high-throughput transcriptome sequencing, we discovered 11 diTPS from jack pine (Pinus banksiana) and lodgepole pine (Pinus contorta). Three of these were orthologous to known conifer bifunctional levopimaradiene/abietadiene synthases. Surprisingly, two sets of orthologous PbdiTPSs and PcdiTPSs were monofunctional class I enzymes that lacked functional class II active sites and converted (+)-copalyl diphosphate, but not GGPP, into isopimaradiene and pimaradiene as major products. Diterpene profiles and transcriptome sequences of lodgepole pine and jack pine are consistent with roles for these diTPSs in DRA biosynthesis. The monofunctional class I diTPSs of DRA biosynthesis form a new clade within the gymnosperm-specific TPS-d3 subfamily that evolved from bifunctional diTPS rather than monofunctional enzymes (TPS-c and TPS-e) of gibberellin metabolism. Homology modeling suggested alterations in the class I active site that may have contributed to their functional specialization relative to other conifer diTPSs.

  17. Terpenes as Green Solvents for Extraction of Oil from Microalgae

    Directory of Open Access Journals (Sweden)

    Celine Dejoye Tanzi

    2012-07-01

    Full Text Available Herein is described a green and original alternative procedure for the extraction of oil from microalgae. Extractions were carried out using terpenes obtained from renewable feedstocks as alternative solvents instead of hazardous petroleum solvents such as n-hexane. The described method is achieved in two steps using Soxhlet extraction followed by the elimination of the solvent from the medium using Clevenger distillation in the second step. Oils extracted from microalgae were compared in terms of qualitative and quantitative determination. No significant difference was obtained between each extract, allowing us to conclude that the proposed method is green, clean and efficient.

  18. Transfer of terpenes from essential oils into cow milk

    DEFF Research Database (Denmark)

    Lejonklev, J.; Løkke, M.M.; Larsen, M.K.

    2013-01-01

    The objective of this study was to investigate the transfer of volatile terpenes from caraway seed and oregano plant essential oils into cow's milk through respiratory and gastrointestinal exposure. Essential oils have potential applications as feed additives because of their antimicrobial...... properties, but very little work exists on the transfer of their volatile compounds into milk. Lactating Danish Holstein cows with duodenum cannula were used. Gastrointestinal exposure was facilitated by infusing the essential oils, mixed with deodorized sesame oil, into the duodenum cannula. Two levels were...

  19. Terpenes of Salvia species leaf oils: chemosystematic implications

    OpenAIRE

    Coassini Lokar, Laura; Moneghini, Mariarosa

    2017-01-01

    Wild specimens of Salvia L. were collected in three different moments of anthesis and their volatile leaf oils were analyzed by GC/GCMS. The quantitative terpene composition is very variable with the anthesis. S. bertolonii is the richest species in a-thujone. S. officinalis is characterized by high percentages of 1,8 cineole, 4-terpineol, isorboneol and a -bisabolol. In S. verticillata high percentages of borneol and {3-cariophyllene are present. In the three species a-thujone was always mor...

  20. KOMPARASI MODEL PEMBELAJARAN TPS DAN SFE TERHADAP KEMAMPUAN PEMAHAMAN KONSEP

    Directory of Open Access Journals (Sweden)

    Febri Susanto

    2014-03-01

    Full Text Available Penelitian ini bertujuan untuk mengetahui apakah pembelajaran model Think Pair Share (TPS dan model Student Facilitator and Explaining (SFE pada siswa kelas X materi jarak pada bangun ruang dapat mencapai ketuntasan belajar dan untuk mengetahui manakah yang lebih efektif antara pembelajaran model Think Pair Share (TPS dan model Student Facilitator and Explaining (SFE. Populasi dalam penelitian ini adalah siswa kelas X SMA N 1 Kota Tegal tahun pelajaran 2012/2013 yang berada dalam sembilan kelas. Sembilan kelas tersebut kemudian dipilih dua kelas secara acak untuk dijadikan sampel dalam penelitian ini yaitu kelas X 2 sebagai kelas eksperimen 1 dan kelas X 1 sebagai kelas eksperimen 2. Uji ketuntasan belajar memberikan hasil yaitu siswa kelas eksperimen 1 dan 2 telah mencapai ketuntasan belajar. Uji kesamaan dua proporsi memberikan hasil yakni proporsi ketuntasan belajar pada aspek pemahaman konsep siswa kelas eksperimen 1 sama baiknya dibanding proporsi ketuntasan belajar pada aspek pemahaman konsep siswa kelas eksperimen 2. Hasil Penelitian menunjukkan pembelajaran model TPS dan SFE pada materi jarak pada bangun ruang telah mencapai ketuntasan belajar dan model pembelajaran TPS sama efektifnya dengan pembelajaran model SFE.

  1. Thermal, mechanical and morphological behavior of starch thermoplastic (TPS) and polycaprolactone (PCL)

    International Nuclear Information System (INIS)

    Campos, Adriana de; Marconcini, Jose M.; Mattoso, Luiz H.C.

    2011-01-01

    Thermal, mechanical and morphological properties of thermoplastic starch (TPS) and polycaprolactone (PCL) blend obtained by extrusion was studied. The results showed that TPS/PCL blends are immiscible, however it is suggested some interaction in the interphase between TPS and PCL as observed by crystallinity decrease of the blend. The PCL addition in the TPS improves the properties and decreases the cost of the blend. (author)

  2. High-temperature behavior of advanced spacecraft TPS

    Science.gov (United States)

    Pallix, Joan

    1994-05-01

    The objective of this work has been to develop more efficient, lighter weight, and higher temperature thermal protection systems (TPS) for future reentry space vehicles. The research carried out during this funding period involved the design, analysis, testing, fabrication, and characterization of thermal protection materials to be used on future hypersonic vehicles. This work is important for the prediction of material performance at high temperature and aids in the design of thermal protection systems for a number of programs including programs such as the National Aerospace Plane (NASP), Pegasus and Pegasus/SWERVE, the Comet Rendezvous and Flyby Vehicle (CRAF), and the Mars mission entry vehicles. Research has been performed in two main areas including development and testing of thermal protection systems (TPS) and computational research. A variety of TPS materials and coatings have been developed during this funding period. Ceramic coatings were developed for flexible insulations as well as for low density ceramic insulators. Chemical vapor deposition processes were established for the fabrication of ceramic matrix composites. Experimental testing and characterization of these materials has been carried out in the NASA Ames Research Center Thermophysics Facilities and in the Ames time-of-flight mass spectrometer facility. By means of computation, we have been better able to understand the flow structure and properties of the TPS components and to estimate the aerothermal heating, stress, ablation rate, thermal response, and shape change on the surfaces of TPS. In addition, work for the computational surface thermochemistry project has included modification of existing computer codes and creating new codes to model material response and shape change on atmospheric entry vehicles in a variety of environments (e.g., earth and Mars atmospheres).

  3. Negative inotropism of terpenes on guinea pig left atrium: structure-activity relationships.

    Science.gov (United States)

    Vasconcelos, Carla M L; Oliveira, Ingrid S N; Santos, José N A; Souza, Américo A; Menezes-Filho, José E R; Silva Neto, Júlio A; Lima, Tamires C; de Sousa, Damião P

    2018-06-01

    The aim of this work was to evaluate the pharmacological effect of seven structurally related terpenes on the contractility of cardiac muscle. The effect of terpenes was studied on isolated electrically driven guinea pig left atrium. From concentration-response curves for inotropic effect were determined the EC 50 and relative potency of such terpenes. Our results revealed that all terpenes, except phytol, showed ability to reduce the contractile response of guinea pig left atrium. Further, relative potency was directly related to the number of isoprene units and to the lipophilicity of the compounds. For example, sesquiterpenes farnesol and nerolidol showed higher relative potency when compared with the monoterpenes citronellol, geraniol and nerol. We can conclude that most of the evaluated terpenes showed a promising negative inotropism on the atrial muscle. Future studies are necessary to investigate their action mechanism.

  4. Mutational, Phylogeny and Evolution Analyses of Salvia Copalyl Diphosphate Synthase

    International Nuclear Information System (INIS)

    Hao, D. C.; Thimmappa, R. B.; Xiao, P. G.

    2016-01-01

    The cyclization of geranylgeranyl diphosphate (GGPP) is catalyzed by copalyl diphosphate synthase (CPS), a class II diterpene synthase (diTPS), to form copalyl diphosphate (CPP), which is an essential substrate of a variety of diterpenes in secondary metabolism of angiosperm including Salvia medicinal plants. The protein environment of the N-terminal class II active site stabilizes the carbocation intermediates and maintains the catalytic activity of angiosperm class II diTPS. The virtual modeling and mutagenesis of the class II diTPS of Salvia miltiorrhiza (SmCPS) were accomplished to illuminate the catalytic activity of SmCPS. Terminal truncations and point mutations established the role of the Beta-Gamma domain and Alpha domain, i.e., they facilitate the flexible conformational change of the class II active site after substrate binding. E203 and K238 in the N-ter Gamma domain of SmCPS1 are functional in the substrate binding and conformational transition and might be essential in catalysis. Similar to other CPSs, the ensuing protonation of the GGPP substrate and coordination of the diphosphate group are governed by highly conserved residues in the DxDD motif of SmCPS, e.g., D372 of CPS1. Moreover, F256 and Y505 stabilize the carbocation and control the enzymatic activity during CPP formation. The amino acids of the predicted active sites, despite under purifying selection, vary greatly, corresponding to the functional flexibility of angiosperm CPSs. Molecular phylogeny and evolution analyses suggest early and ongoing evolution of labdane-related diterpenoid metabolism in angiosperm. (author)

  5. Functional Characterization of Sesquiterpene Synthase from Polygonum minus

    Directory of Open Access Journals (Sweden)

    Su-Fang Ee

    2014-01-01

    Full Text Available Polygonum minus is an aromatic plant, which contains high abundance of terpenoids, especially the sesquiterpenes C15H24. Sesquiterpenes were believed to contribute to the many useful biological properties in plants. This study aimed to functionally characterize a full length sesquiterpene synthase gene from P. minus. P. minus sesquiterpene synthase (PmSTS has a complete open reading frame (ORF of 1689 base pairs encoding a 562 amino acid protein. Similar to other sesquiterpene synthases, PmSTS has two large domains: the N-terminal domain and the C-terminal metal-binding domain. It also consists of three conserved motifs: the DDXXD, NSE/DTE, and RXR. A three-dimensional protein model for PmSTS built clearly distinguished the two main domains, where conserved motifs were highlighted. We also constructed a phylogenetic tree, which showed that PmSTS belongs to the angiosperm sesquiterpene synthase subfamily Tps-a. To examine the function of PmSTS, we expressed this gene in Arabidopsis thaliana. Two transgenic lines, designated as OE3 and OE7, were further characterized, both molecularly and functionally. The transgenic plants demonstrated smaller basal rosette leaves, shorter and fewer flowering stems, and fewer seeds compared to wild type plants. Gas chromatography-mass spectrometry analysis of the transgenic plants showed that PmSTS was responsible for the production of β-sesquiphellandrene.

  6. Induction of Terpene Biosynthesis in Berries of Microvine Transformed with VvDXS1 Alleles

    Directory of Open Access Journals (Sweden)

    Lorenza Dalla Costa

    2018-01-01

    Full Text Available Terpenoids, especially monoterpenes, are major aroma-impact compounds in grape and wine. Previous studies highlighted a key regulatory role for grapevine 1-deoxy-D-xylulose 5-phosphate synthase 1 (VvDXS1, the first enzyme of the methylerythritol phosphate pathway for isoprenoid precursor biosynthesis. Here, the parallel analysis of VvDXS1 genotype and terpene concentration in a germplasm collection demonstrated that VvDXS1 sequence has a very high predictive value for the accumulation of monoterpenes and also has an influence on sesquiterpene levels. A metabolic engineering approach was applied by expressing distinct VvDXS1 alleles in the grapevine model system “microvine” and assessing the effects on downstream pathways at transcriptional and metabolic level in different organs and fruit developmental stages. The underlying goal was to investigate two potential perturbation mechanisms, the former based on a significant over-expression of the wild-type (neutral VvDXS1 allele and the latter on the ex-novo expression of an enzyme with increased catalytic efficiency from the mutated (muscat VvDXS1 allele. The integration of the two VvDXS1 alleles in distinct microvine lines was found to alter the expression of several terpenoid biosynthetic genes, as assayed through an ad hoc developed TaqMan array based on cDNA libraries of four aromatic cultivars. In particular, enhanced transcription of monoterpene, sesquiterpene and carotenoid pathway genes was observed. The accumulation of monoterpenes in ripe berries was higher in the transformed microvines compared to control plants. This effect is predominantly attributed to the improved activity of the VvDXS1 enzyme coded by the muscat allele, whereas the up-regulation of VvDXS1 plays a secondary role in the increase of monoterpenes.

  7. Functional identification of a Lippia dulcis bornyl diphosphate synthase that contains a duplicated, inhibitory arginine-rich motif.

    Science.gov (United States)

    Hurd, Matthew C; Kwon, Moonhyuk; Ro, Dae-Kyun

    2017-08-26

    Lippia dulcis (Aztec sweet herb) contains the potent natural sweetener hernandulcin, a sesquiterpene ketone found in the leaves and flowers. Utilizing the leaves for agricultural application is challenging due to the presence of the bitter-tasting and toxic monoterpene, camphor. To unlock the commercial potential of L. dulcis leaves, the first step of camphor biosynthesis by a bornyl diphosphate synthase needs to be elucidated. Two putative monoterpene synthases (LdTPS3 and LdTPS9) were isolated from L. dulcis leaf cDNA. To elucidate their catalytic functions, E. coli-produced recombinant enzymes with truncations of their chloroplast transit peptides were assayed with geranyl diphosphate (GPP). In vitro enzyme assays showed that LdTPS3 encodes bornyl diphosphate synthase (thus named LdBPPS) while LdTPS9 encodes linalool synthase. Interestingly, the N-terminus of LdBPPS possesses two arginine-rich (RRX 8 W) motifs, and enzyme assays showed that the presence of both RRX 8 W motifs completely inhibits the catalytic activity of LdBPPS. Only after the removal of the putative chloroplast transit peptide and the first RRX 8 W, LdBPPS could react with GPP to produce bornyl diphosphate. LdBPPS is distantly related to the known bornyl diphosphate synthase from sage in a phylogenetic analysis, indicating a converged evolution of camphor biosynthesis in sage and L. dulcis. The discovery of LdBPPS opens up the possibility of engineering L. dulcis to remove the undesirable product, camphor. Copyright © 2017 Elsevier Inc. All rights reserved.

  8. Conformal Ablative Thermal Protection Systems (CA-TPS) for Venus and Saturn Backshells

    Science.gov (United States)

    Beck, R.; Gasch, M.; Stackpoole, M.; Wilder, M.; Boghozian, T.; Chavez-Garcia, J.; Prabhu, Dinesh; Kazemba, Cole D.; Venkatapathy, E.

    2016-01-01

    This poster provides an overview of the work performed to date on the Conformal Ablative TPS (CA-TPS) element of the TPSM project out of GCDP. Under this element, NASA is developing improved ablative TPS materials based on flexible felt for reinforcement rather than rigid reinforcements. By replacing the reinforcements with felt, the resulting materials have much higher strain-to-failure and are much lower in thermal conductivity than their rigid counterparts. These characteristics should allow for larger tile sizes, direct bonding to aeroshells and even lower weight TPS. The conformal phenolic impregnated carbon felt (C-PICA) is a candidate for backshell TPS for both Venus and Saturn entry vehicles.

  9. Structural and In Vivo Studies on Trehalose-6-Phosphate Synthase from Pathogenic Fungi Provide Insights into Its Catalytic Mechanism, Biological Necessity, and Potential for Novel Antifungal Drug Design

    Energy Technology Data Exchange (ETDEWEB)

    Miao, Yi; Tenor, Jennifer L.; Toffaletti, Dena L.; Maskarinec, Stacey A.; Liu, Jiuyu; Lee, Richard E.; Perfect, John R.; Brennan, Richard G.; Hendrickson, Wayne A.

    2017-07-25

    ABSTRACT

    The disaccharide trehalose is critical to the survival of pathogenic fungi in their human host. Trehalose-6-phosphate synthase (Tps1) catalyzes the first step of trehalose biosynthesis in fungi. Here, we report the first structures of eukaryotic Tps1s in complex with substrates or substrate analogues. The overall structures of Tps1 fromCandida albicansandAspergillus fumigatusare essentially identical and reveal N- and C-terminal Rossmann fold domains that form the glucose-6-phosphate and UDP-glucose substrate binding sites, respectively. These Tps1 structures with substrates or substrate analogues reveal key residues involved in recognition and catalysis. Disruption of these key residues severely impaired Tps1 enzymatic activity. Subsequent cellular analyses also highlight the enzymatic function of Tps1 in thermotolerance, yeast-hypha transition, and biofilm development. These results suggest that Tps1 enzymatic functionality is essential for the fungal stress response and virulence. Furthermore, structures of Tps1 in complex with the nonhydrolyzable inhibitor, validoxylamine A, visualize the transition state and support an internal return-like catalytic mechanism that is generalizable to other GT-B-fold retaining glycosyltransferases. Collectively, our results depict key Tps1-substrate interactions, unveil the enzymatic mechanism of these fungal proteins, and pave the way for high-throughput inhibitor screening buttressed and guided by the current structures and those of high-affinity ligand-Tps1 complexes.

    IMPORTANCEInvasive fungal diseases have emerged as major threats, resulting in more than 1.5 million deaths annually worldwide. This epidemic has been further complicated by increasing resistance to all major classes of antifungal drugs in the clinic. Trehalose biosynthesis is essential for the fungal stress response and virulence. Critically, this biosynthetic pathway is absent in

  10. HASIL BELAJAR DAN PEMBELAJARAN KOOPERATIF TIPE THINK PAIR SHARE (TPS

    Directory of Open Access Journals (Sweden)

    Loli Jasdilla

    2017-03-01

    Full Text Available The objectives of this research is to improve social science outcomes by using Think Pair Share (TPS for the fourth grade in elementary school. This research is classroom action research. The procedure of this research conducted planning, acting, observation, and reflection. Subject of this research was 22 students for the fourth grade in SD 20 Kalumbuk, Padang. Techniques of collecting data were using test and observation. Data analyzed by using thematic analysis. Data validated by using triangulation, member checking, and reflecivity. The students’ learning outcomes improved from 76,75% into 89,67% (very good. The result of this research shows that cooperative model of Think Pair Share (TPS type can improve social science outcomes for the fourth grade students of elementary school.

  11. Terpene chemodiversity of relict conifers Picea omorika, Pinus heldreichii, and Pinus peuce, endemic to Balkan.

    Science.gov (United States)

    Nikolić, Biljana; Ristić, Mihailo; Tešević, Vele; Marin, Petar D; Bojović, Srdjan

    2011-12-01

    Terpenes are often used as ecological and chemotaxonomic markers of plant species, as well as for estimation of geographic variability. Essential oils of relic and Balkan endemic/subendemic conifers, Picea omorika, Pinus heldreichii, and P. peuce, in central part of Balkan Peninsula (Serbia and Montenegro), on the level of terpene classes and common terpene compounds were investigated. In finding terpene combinations, which could show the best diversity between species and their natural populations, several statistical methods were applied. Apart from the content of different terpene classes (P. omorika has the most abundant O-containing monoterpenes and sesquiterpenes; P. heldreichii and P. peuce have the largest abundance of sesquiterpene and monoterpene hydrocarbons, resp.), the species are clearly separated according to terpene profile with 22 common compounds. But, divergences in their populations were established only in combination of several compounds (specific for each species), and they were found to be the results of geomorphologic, climatic, and genetic factors. We found similarities between investigated species and some taxa from literature with respect to terpene composition, possibly due to hybridization and phylogenetic relations. Obtained results are also important regarding to chemotaxonomy, biogeography, phylogeny, and evolution of these taxa. Copyright © 2011 Verlag Helvetica Chimica Acta AG, Zürich.

  12. Small RNA Transcriptome of Hibiscus Syriacus Provides Insights into the Potential Influence of microRNAs in Flower Development and Terpene Synthesis.

    Science.gov (United States)

    Kim, Taewook; Park, June Hyun; Lee, Sang-Gil; Kim, Soyoung; Kim, Jihyun; Lee, Jungho; Shin, Chanseok

    2017-08-01

    MicroRNAs (miRNAs) are essential small RNA molecules that regulate the expression of target mRNAs in plants and animals. Here, we aimed to identify miRNAs and their putative targets in Hibiscus syriacus , the national flower of South Korea. We employed high-throughput sequencing of small RNAs obtained from four different tissues ( i.e. , leaf, root, flower, and ovary) and identified 33 conserved and 30 novel miRNA families, many of which showed differential tissue-specific expressions. In addition, we computationally predicted novel targets of miRNAs and validated some of them using 5' rapid amplification of cDNA ends analysis. One of the validated novel targets of miR477 was a terpene synthase, the primary gene involved in the formation of disease-resistant terpene metabolites such as sterols and phytoalexins. In addition, a predicted target of conserved miRNAs, miR396, is SHORT VEGETATIVE PHASE , which is involved in flower initiation and is duplicated in H. syriacus . Collectively, this study provides the first reliable draft of the H. syriacus miRNA transcriptome that should constitute a basis for understanding the biological roles of miRNAs in H. syriacus.

  13. Determination of the terpene flux from orange species and Norway spruce by relaxed eddy accumulation

    DEFF Research Database (Denmark)

    Christensen, C.S.; Hummelshøj, P.; Jensen, N.O.

    2000-01-01

    Terpene fluxes from a Norway spruce (Picea abies) forest and an orange orchard (Citrus clementii and Citrus sinensis) were measured by relaxed eddy accumulation (REA) during summer 1997. alpha-pinene and beta-pinene were the most abundant terpenes emitted from Norway spruce and constituted approx...... rate by using two precision pumps operated at approximately 60 mi min(-1). The terpenes collected on the adsorbent tubes were significantly decomposed by ozone during sampling unless ozone scrubbers were applied. (C) 2000 Elsevier Science Ltd. All rights reserved....

  14. Expression of TPS1 gene from Saccharomycopsis fibuligera A11 in Saccharomyces sp. W0 enhances trehalose accumulation, ethanol tolerance, and ethanol production.

    Science.gov (United States)

    Cao, Tian-Shu; Chi, Zhe; Liu, Guang-Lei; Chi, Zhen-Ming

    2014-01-01

    It has been reported that trehalose plays an important role in stress tolerance in yeasts. Therefore, in order to construct a stably recombinant Saccharomyces sp. W0 with higher ethanol tolerance, the TPS1 gene encoding 6-phosphate-trehalose synthase cloned from Saccharomycopsis fibuligera A11 was ligated into the 18S rDNA integration vector pMIRSC11 and integrated into chromosomal DNA of Saccharomyces sp. W0. The transformant Z8 obtained had the content of 6.23 g of trehalose/100 g of cell dry weight, while Saccharomyces sp. W0 only contained 4.05 g of trehalose/100 g of cell dry weight. The transformant Z8 also had higher ethanol tolerance (cell survival was 25.1 % at 18 ml of ethanol/100 ml of solution) and trehalose-6-phosphate synthase (Tps1) activity (1.3 U/mg) and produced more ethanol (16.4 ml of ethanol/100 ml of medium) than Saccharomyces sp. W0 (cell survival was 12.1 % at 18 ml of ethanol/100 ml of solution, Tps1 activity was 0.8 U/mg and the produced ethanol concentration was 14.2 ml of ethanol/100 ml of medium) under the same conditions. The results show that trehalose indeed can play an important role in ethanol tolerance and ethanol production by Saccharomyces sp. W0.

  15. Indoor fine particles: the role of terpene emissions from consumer products.

    Science.gov (United States)

    Sarwar, Golam; Olson, David A; Corsi, Richard L; Weschler, Charles J

    2004-03-01

    Consumer products can emit significant quantities of terpenes, which can react with ozone (O3). Resulting byproducts include compounds with low vapor pressures that contribute to the growth of secondary organic aerosols (SOAs). The focus of this study was to evaluate the potential for SOA growth, in the presence of O3, following the use of a lime-scented liquid air freshener, a pine-scented solid air freshener, a lemon-scented general-purpose cleaner, a wood floor cleaner, and a perfume. Two chamber experiments were performed for each of these five terpene-containing agents, one at an elevated O3 concentration and-the other at a lower O3 concentration. Particle number and mass concentrations increased and O3 concentrations decreased during each experiment. Experiments with terpene-based air fresheners produced the highest increases in particle number and mass concentrations. The results of this study clearly demonstrate that homogeneous reactions between O3 and terpenes from various consumer products can lead to increases in fine particle mass concentrations when these products are used indoors. Particle increases can occur during periods of elevated outdoor O3 concentrations or indoor O3 generation, coupled with elevated terpene releases. Human exposure to fine particles can be reduced by minimizing indoor terpene concentrations or O3 concentrations.

  16. Chemodiversity in terpene emissions at a boreal Scots pine stand

    Science.gov (United States)

    Bäck, J.; Aalto, J.; Henriksson, M.; Hakola, H.; He, Q.; Boy, M.

    2011-10-01

    Atmospheric chemistry in background areas is strongly influenced by natural vegetation. Coniferous forests are known to produce large quantities of volatile vapors, especially terpenes to the surrounding air. These compounds are reactive in the atmosphere, and contribute to the formation and growth of atmospheric new particles. Our aim was to analyze the variability of mono- and sesquiterpene emissions between Scots pine trees, in order to clarify the potential errors caused by using emission data obtained from only a few trees in atmospheric chemistry models. We also aimed at testing if stand history and seed origin has an influence on the chemotypic diversity. The inherited, chemotypic variability in mono- and sesquiterpene emission was studied in a seemingly homogeneous 47-yr-old stand in Southern Finland, where two areas differing in their stand regeneration history could be distinguished. Sampling was conducted in August 2009. Terpene concentrations in the air had been measured at the same site for seven years prior to branch sampling for chemotypes. Two main compounds, α-pinene and Δ3-carene formed together 40-97% of the monoterpene proportions in both the branch emissions and in the air concentrations. The data showed a bimodal distribution in emission composition, in particular in Δ3-carene emission within the studied population. 10% of the trees emitted mainly α-pinene and no Δ3-carene at all, whereas 20% of the trees where characterized as high Δ3-carene emitters (Δ3-carene forming >80% of total emitted monoterpene spectrum). An intermediate group of trees emitted equal amounts of both α-pinene and Δ3-carene. The emission pattern of trees at the area established using seeding as the artificial regeneration method differed from the naturally regenerated or planted trees, being mainly high Δ3-carene emitters. Some differences were also seen in e.g. camphene and limonene emissions between chemotypes, but sesquiterpene emissions did not differ

  17. Compensator design for corrector magnet power supply of TPS facility

    International Nuclear Information System (INIS)

    Wong, Y.-S.; Chen, J.-F.; Liu, K.-B.; Liu, C.-Y.; Wang, B.-S.

    2017-01-01

    From 2012 to 2015, Taiwan government has a most important technology project is Taiwan Photon Source (TPS), the total budget of TPS fund to over US300 million. It set up a synchrotron storage ring (electron energy of 3.3 GeV, circumference of 518 m, and low emittance) that provides one of the world's brightest synchrotron sources of x-rays. This study presents a compensator design for corrector magnet power supply to avoid limitations in stabilizing the frequency when the machine output current load is valid. A lead-lag compensator had been built in a full-bridge converter to improve the system bandwidth. Lead-lag compensators influence various disciplines, such as robotics, satellite control, automobile diagnostics, and laser frequency stabilization. These components are important building blocks in analog control systems and can also be used in digital control. A 50V output voltage and 10A output current prototype converter is fabricated in the laboratory. From the experimental results, the effectiveness of the control loop design can be verified from the gain margin and phase margin.

  18. Compensator design for corrector magnet power supply of TPS facility

    Science.gov (United States)

    Wong, Y.-S.; Chen, J.-F.; Liu, K.-B.; Liu, C.-Y.; Wang, B.-S.

    2017-10-01

    From 2012 to 2015, Taiwan government has a most important technology project is Taiwan Photon Source (TPS), the total budget of TPS fund to over US300 million. It set up a synchrotron storage ring (electron energy of 3.3 GeV, circumference of 518 m, and low emittance) that provides one of the world's brightest synchrotron sources of x-rays. This study presents a compensator design for corrector magnet power supply to avoid limitations in stabilizing the frequency when the machine output current load is valid. A lead-lag compensator had been built in a full-bridge converter to improve the system bandwidth. Lead-lag compensators influence various disciplines, such as robotics, satellite control, automobile diagnostics, and laser frequency stabilization. These components are important building blocks in analog control systems and can also be used in digital control. A 50V output voltage and 10A output current prototype converter is fabricated in the laboratory. From the experimental results, the effectiveness of the control loop design can be verified from the gain margin and phase margin.

  19. Biosynthesis of the phenolic monoterpenes , thymol and carvacrol , by terpene synthases and cytochrome P450s in oregano and thyme

    DEFF Research Database (Denmark)

    Crocoll, Christoph

    2011-01-01

    Plants are sophisticated light-driven “green” factories able to synthesize an immense number of bio-active natural products (Jensen and Møller, 2010). These natural products are also referred to as secondary products or secondary metabolites since they are not directly essential for the basic pro...

  20. Hybrid polyketide synthases

    Energy Technology Data Exchange (ETDEWEB)

    Fortman, Jeffrey L.; Hagen, Andrew; Katz, Leonard; Keasling, Jay D.; Poust, Sean; Zhang, Jingwei; Zotchev, Sergey

    2016-05-10

    The present invention provides for a polyketide synthase (PKS) capable of synthesizing an even-chain or odd-chain diacid or lactam or diamine. The present invention also provides for a host cell comprising the PKS and when cultured produces the even-chain diacid, odd-chain diacid, or KAPA. The present invention also provides for a host cell comprising the PKS capable of synthesizing a pimelic acid or KAPA, and when cultured produces biotin.

  1. Identification and characterization of two bisabolene synthases from linear glandular trichomes of sunflower (Helianthus annuus L., Asteraceae).

    Science.gov (United States)

    Aschenbrenner, Anna-Katharina; Kwon, Moonhyuk; Conrad, Jürgen; Ro, Dae-Kyun; Spring, Otmar

    2016-04-01

    Sunflower is known to produce a variety of bisabolene-type sesquiterpenes and accumulates these substances in trichomes of leaves, stems and flowering parts. A bioinformatics approach was used to identify the enzyme responsible for the initial step in the biosynthesis of these compounds from its precursor farnesyl pyrophosphate. Based on sequence similarity with a known bisabolene synthases from Arabidopsis thaliana AtTPS12, candidate genes of Helianthus were searched in EST-database and used to design specific primers. PCR experiments identified two candidates in the RNA pool of linear glandular trichomes of sunflower. Their sequences contained the typical motifs of sesquiterpene synthases and their expression in yeast functionally characterized them as bisabolene synthases. Spectroscopic analysis identified the stereochemistry of the product of both enzymes as (Z)-γ-bisabolene. The origin of the two sunflower bisabolene synthase genes from the transcripts of linear trichomes indicates that they may be involved in the synthesis of sesquiterpenes produced in these trichomes. Comparison of the amino acid sequences of the sunflower bisabolene synthases showed high similarity with sesquiterpene synthases from other Asteracean species and indicated putative evolutionary origin from a β-farnesene synthase. Copyright © 2016 Elsevier Ltd. All rights reserved.

  2. Application of extrusion-cooking for processing of thermoplastic starch (TPS)

    NARCIS (Netherlands)

    Moscicki, Leszek; Mitrus, Marcin; Wojtowicz, Agnieszka; Oniszczuk, Tomasz; Rejak, Andrzej; Janssen, Leon; Mościcki, Leszek

    Thermoplastic starch (TPS) as fully biodegradable biopolymer appeared to be one of the most useful and promising materials for packaging purpose. To obtain TPS thermal and mechanical processing should disrupt semi-crystalline starch granules. As the melting temperature of pure starch is

  3. Characterizing TPS Microstructure: A Review of Some techniques

    Science.gov (United States)

    Gasch, Matthew; Stackpole, Mairead; Agrawal, Parul; Chavez-Garcie, Jose

    2011-01-01

    I. When seeking to understand ablator microstructure and morphology there are several useful techniques A. SEM 1) Visual characteriza3on at various length scales. 2) Chemical mapping by backscatter or x-ray highlights areas of interest. 3) Combined with other techniques (density, weight change, chemical analysis) SEM is a powerful tool to aid in explaining thermo/structural data. B. ASAP. 1) Chemical characteriza3on at various length scales. 2) Chemical mapping of pore structure by gas adsorption. 3) Provides a map of pore size vs. pore volume. 4) Provided surface area of exposed TPS. II. Both methods help characterize and understand how ablators react with other chemical species and provides insight into how they oxidize.

  4. Dose evaluation of TPS according to treatment sites in IMRT

    International Nuclear Information System (INIS)

    Kim, Jin Man; Kim, Jong Sik; Hong, Chae Seon; Park, Ju Young; Park, Su Yeon; Ju, Sang Gyu

    2013-01-01

    This study executed therapy plans on prostate cancer (homogeneous density area) and lung cancer (non-homogeneous density area) using radiation treatment planning systems such as Pinnacle 3 (version 9.2, Philips Medical Systems, USA) and Eclipse (version 10.0, Varian Medical Systems, USA) in order to quantify the difference between dose calculation according to density in IMRT. The subjects were prostate cancer patients (n=5) and lung cancer patients (n=5) who had therapies in our hospital. Identical constraints and optimization process according to the Protocol were administered on the subjects. For the therapy plan of prostate cancer patients, 10 MV and 7Beam were used and 2.5 Gy was prescribed in 28 fx to make 70 Gy in total. For lung cancer patients, 6 MV and 6Beam were used and 2 Gy was prescribed in 33 fx to make 66 Gy in total. Through two therapy planning systems, maximum dose, average dose, and minimum dose of OAR (Organ at Risk) of CTV, PTV and around tumor were investigated. In prostate cancer, both therapy planning systems showed within 2% change of dose of CTV and PTV and normal organs (Bladder, Both femur and Rectum out) near the tumor satisfied the dose constraints. In lung cancer, CTV and PTV showed less than 2% changes in dose and normal organs (Esophagus, Spinal cord and Both lungs) satisfied dose restrictions. However, the minimum dose of Eclipse therapy plan was 1.9% higher in CTV and 3.5% higher in PTV, and in case of both lungs there was 3.0% difference at V5 Gy. Each TPS according to the density satisfied dose limits of our hospital proving the clinical accuracy. It is considered more accurate and precise therapy plan can be made if studies on treatment planning for diverse parts and the application of such TPS are made

  5. Molecular Architecture and Biomedical Leads of Terpenes from Red Sea Marine Invertebrates

    Science.gov (United States)

    Hegazy, Mohamed Elamir F.; Mohamed, Tarik A.; Alhammady, Montaser A.; Shaheen, Alaa M.; Reda, Eman H.; Elshamy, Abdelsamed I.; Aziz, Mina; Paré, Paul W.

    2015-01-01

    Marine invertebrates including sponges, soft coral, tunicates, mollusks and bryozoan have proved to be a prolific source of bioactive natural products. Among marine-derived metabolites, terpenoids have provided a vast array of molecular architectures. These isoprenoid-derived metabolites also exhibit highly specialized biological activities ranging from nerve regeneration to blood-sugar regulation. As a result, intense research activity has been devoted to characterizing invertebrate terpenes from both a chemical and biological standpoint. This review focuses on the chemistry and biology of terpene metabolites isolated from the Red Sea ecosystem, a unique marine biome with one of the highest levels of biodiversity and specifically rich in invertebrate species. PMID:26006713

  6. Molecular Architecture and Biomedical Leads of Terpenes from Red Sea Marine Invertebrates

    Directory of Open Access Journals (Sweden)

    Mohamed Elamir F. Hegazy

    2015-05-01

    Full Text Available Marine invertebrates including sponges, soft coral, tunicates, mollusks and bryozoan have proved to be a prolific source of bioactive natural products. Among marine-derived metabolites, terpenoids have provided a vast array of molecular architectures. These isoprenoid-derived metabolites also exhibit highly specialized biological activities ranging from nerve regeneration to blood-sugar regulation. As a result, intense research activity has been devoted to characterizing invertebrate terpenes from both a chemical and biological standpoint. This review focuses on the chemistry and biology of terpene metabolites isolated from the Red Sea ecosystem, a unique marine biome with one of the highest levels of biodiversity and specifically rich in invertebrate species.

  7. Temporal effects of prescribed burning on terpene production in Mediterranean pines.

    Science.gov (United States)

    Valor, Teresa; Ormeño, Elena; Casals, Pere

    2017-12-01

    Prescribed burning is used to reduce fuel hazard but underburning can damage standing trees. The effect of burning on needle terpene storage, a proxy for secondary metabolism, in fire-damaged pines is poorly understood despite the protection terpenes confer against biotic and abiotic stressors. We investigated variation in needle terpene storage after burning in three Mediterranean pine species featuring different adaptations to fire regimes. In two pure-stands of Pinus halepensis Mill. and two mixed-stands of Pinus sylvestris L. and Pinus nigra ssp. salzmanni (Dunal) Franco, we compared 24 h and 1 year post-burning concentrations with pre-burning concentrations in 20 trees per species, and evaluated the relative contribution of tree fire severity and physiological condition (δ13C and N concentration) on temporal terpene dynamics (for mono- sesqui- and diterpenes). Twenty-four hours post-burning, monoterpene concentrations were slightly higher in P. halepensis than at pre-burning, while values were similar in P. sylvestris. Differently, in the more fire-resistant P. nigra monoterpene concentrations were lower at 24 h, compared with pre-burning. One year post-burning, concentrations were always lower compared with pre- or 24 h post-burning, regardless of the terpene group. Mono- and sesquiterpene variations were negatively related to pre-burning δ13C, while diterpene variations were associated with fire-induced changes in needle δ13C and N concentration. At both post-burning times, mono- and diterpene concentrations increased significantly with crown scorch volume in all species. Differences in post-burning terpene contents as a function of the pine species' sensitivity to fire suggest that terpenic metabolites could have adaptive importance in fire-prone ecosystems in terms of flammability or defence against biotic agents post-burning. One year post-burning, our results suggest that in a context of fire-induced resource availability, pines likely prioritize

  8. An Arabidopsis callose synthase

    DEFF Research Database (Denmark)

    Ostergaard, Lars; Petersen, Morten; Mattsson, Ole

    2002-01-01

    in the Arabidopsis mpk4 mutant which exhibits systemic acquired resistance (SAR), elevated beta-1,3-glucan synthase activity, and increased callose levels. In addition, AtGsl5 is a likely target of salicylic acid (SA)-dependent SAR, since AtGsl5 mRNA accumulation is induced by SA in wild-type plants, while...... expression of the nahG salicylate hydroxylase reduces AtGsl5 mRNA levels in the mpk4 mutant. These results indicate that AtGsl5 is likely involved in callose synthesis in flowering tissues and in the mpk4 mutant....

  9. Monoterpene synthases from common sage (Salvia officinalis)

    Energy Technology Data Exchange (ETDEWEB)

    Croteau, Rodney Bruce (Pullman, WA); Wise, Mitchell Lynn (Pullman, WA); Katahira, Eva Joy (Pullman, WA); Savage, Thomas Jonathan (Christchurch 5, NZ)

    1999-01-01

    cDNAs encoding (+)-bornyl diphosphate synthase, 1,8-cineole synthase and (+)-sabinene synthase from common sage (Salvia officinalis) have been isolated and sequenced, and the corresponding amino acid sequences has been determined. Accordingly, isolated DNA sequences (SEQ ID No:1; SEQ ID No:3 and SEQ ID No:5) are provided which code for the expression of (+)-bornyl diphosphate synthase (SEQ ID No:2), 1,8-cineole synthase (SEQ ID No:4) and (+)-sabinene synthase SEQ ID No:6), respectively, from sage (Salvia officinalis). In other aspects, replicable recombinant cloning vehicles are provided which code for (+)-bornyl diphosphate synthase, 1,8-cineole synthase or (+)-sabinene synthase, or for a base sequence sufficiently complementary to at least a portion of (+)-bornyl diphosphate synthase, 1,8-cineole synthase or (+)-sabinene synthase DNA or RNA to enable hybridization therewith. In yet other aspects, modified host cells are provided that have been transformed, transfected, infected and/or injected with a recombinant cloning vehicle and/or DNA sequence encoding (+)-bornyl diphosphate synthase, 1,8-cineole synthase or (+)-sabinene synthase. Thus, systems and methods are provided for the recombinant expression of the aforementioned recombinant monoterpene synthases that may be used to facilitate their production, isolation and purification in significant amounts. Recombinant (+)-bornyl diphosphate synthase, 1,8-cineole synthase and (+)-sabinene synthase may be used to obtain expression or enhanced expression of (+)-bornyl diphosphate synthase, 1,8-cineole synthase and (+)-sabinene synthase in plants in order to enhance the production of monoterpenoids, or may be otherwise employed for the regulation or expression of (+)-bornyl diphosphate synthase, 1,8-cineole synthase and (+)-sabinene synthase, or the production of their products.

  10. Verification of Treatment Planning System (TPS) on Beam Axis of Co-60 Teletherapy

    International Nuclear Information System (INIS)

    Nunung-Nuraeni; Budhy-Kurniawan; Purwanto; Sugiyantari; Heru-Prasetio; Nasukha

    2001-01-01

    Cancer diseases up to now can be able to be treated by using surgery, chemotherapy and radiotherapy. The need of high level precision and accuracy on radiation dose are very important task. One of task is verification of Treatment Planning System (Tps) to the treatment of patients. The research has been done to verify Tps on beam exis of teletherapy Co-60. Result found that the different between Tps and measurements are about -2.682 % to 1.918% for simple geometry and homogeneous material, 5.278 % to 4.990 % for complex geometry, and -3.202 % to -2.090 % for more complex geometry. (author)

  11. Bifunctional cis-Abienol Synthase from Abies balsamea Discovered by Transcriptome Sequencing and Its Implications for Diterpenoid Fragrance Production*

    Science.gov (United States)

    Zerbe, Philipp; Chiang, Angela; Yuen, Macaire; Hamberger, Björn; Hamberger, Britta; Draper, Jason A.; Britton, Robert; Bohlmann, Jörg

    2012-01-01

    The labdanoid diterpene alcohol cis-abienol is a major component of the aromatic oleoresin of balsam fir (Abies balsamea) and serves as a valuable bioproduct material for the fragrance industry. Using high-throughput 454 transcriptome sequencing and metabolite profiling of balsam fir bark tissue, we identified candidate diterpene synthase sequences for full-length cDNA cloning and functional characterization. We discovered a bifunctional class I/II cis-abienol synthase (AbCAS), along with the paralogous levopimaradiene/abietadiene synthase and isopimaradiene synthase, all of which are members of the gymnosperm-specific TPS-d subfamily. The AbCAS-catalyzed formation of cis-abienol proceeds via cyclization and hydroxylation at carbon C-8 of a postulated carbocation intermediate in the class II active site, followed by cleavage of the diphosphate group and termination of the reaction sequence without further cyclization in the class I active site. This reaction mechanism is distinct from that of synthases of the isopimaradiene- or levopimaradiene/abietadiene synthase type, which employ deprotonation reactions in the class II active site and secondary cyclizations in the class I active site, leading to tricyclic diterpenes. Comparative homology modeling suggested the active site residues Asp-348, Leu-617, Phe-696, and Gly-723 as potentially important for the specificity of AbCAS. As a class I/II bifunctional enzyme, AbCAS is a promising target for metabolic engineering of cis-abienol production. PMID:22337889

  12. WE-AB-201-03: TPS Commissioning and QA: Incorporating the Entire Planning Process

    International Nuclear Information System (INIS)

    Mutic, S.

    2015-01-01

    Treatment planning systems (TPS) are a cornerstone of modern radiation therapy. Errors in their commissioning or use can have a devastating impact on many patients. To support safe and high quality care, medical physicists must conduct efficient and proper commissioning, good clinical integration, and ongoing quality assurance (QA) of the TPS. AAPM Task Group 53 and related publications have served as seminal benchmarks for TPS commissioning and QA over the past two decades. Over the same time, continuing innovations have made the TPS even more complex and more central to the clinical process. Medical goals are now expressed in terms of the dose and margins around organs and tissues that are delineated from multiple imaging modalities (CT, MR and PET); and even temporally resolved (i.e., 4D) imaging. This information is passed on to optimization algorithms to establish accelerator movements that are programmed directly for IMRT, VMAT and stereotactic treatments. These advances have made commissioning and QA of the TPS much more challenging. This education session reviews up-to-date experience and guidance on this subject; including the recently published AAPM Medical Physics Practice Guideline (MPPG) #5 “Commissioning and QA of Treatment Planning Dose Calculations: Megavoltage Photon and Electron Beams”. Treatment Planning System Commissioning and QA: Challenges and Opportunities (Greg Salomons) This session will provide some key background and review publications describing prominent incidents relating to TPS commissioning and QA. Traditional approaches have been hardware and feature oriented. They aim to establish a functional configuration and establish specifications for regular testing of features (like dose calculation) to assure stable operation and detect failures. With the advent of more complex systems, more patient-specific testing has also been adopted. A number of actual TPS defects will be presented along with heuristics for identifying similar

  13. WE-AB-201-03: TPS Commissioning and QA: Incorporating the Entire Planning Process

    Energy Technology Data Exchange (ETDEWEB)

    Mutic, S. [Washington University School of Medicine (United States)

    2015-06-15

    Treatment planning systems (TPS) are a cornerstone of modern radiation therapy. Errors in their commissioning or use can have a devastating impact on many patients. To support safe and high quality care, medical physicists must conduct efficient and proper commissioning, good clinical integration, and ongoing quality assurance (QA) of the TPS. AAPM Task Group 53 and related publications have served as seminal benchmarks for TPS commissioning and QA over the past two decades. Over the same time, continuing innovations have made the TPS even more complex and more central to the clinical process. Medical goals are now expressed in terms of the dose and margins around organs and tissues that are delineated from multiple imaging modalities (CT, MR and PET); and even temporally resolved (i.e., 4D) imaging. This information is passed on to optimization algorithms to establish accelerator movements that are programmed directly for IMRT, VMAT and stereotactic treatments. These advances have made commissioning and QA of the TPS much more challenging. This education session reviews up-to-date experience and guidance on this subject; including the recently published AAPM Medical Physics Practice Guideline (MPPG) #5 “Commissioning and QA of Treatment Planning Dose Calculations: Megavoltage Photon and Electron Beams”. Treatment Planning System Commissioning and QA: Challenges and Opportunities (Greg Salomons) This session will provide some key background and review publications describing prominent incidents relating to TPS commissioning and QA. Traditional approaches have been hardware and feature oriented. They aim to establish a functional configuration and establish specifications for regular testing of features (like dose calculation) to assure stable operation and detect failures. With the advent of more complex systems, more patient-specific testing has also been adopted. A number of actual TPS defects will be presented along with heuristics for identifying similar

  14. Friction and wear of TPS fibers. Progress Report

    International Nuclear Information System (INIS)

    Bascom, W.D.; Wong, S.

    1987-11-01

    The sliding friction behavior of single filaments of SiO 2 , SiC, and an aluminoborosilicate has been determined. These fibers are used in thermal protection systems (TPS) and are subject to damage during weaving and aero-maneuvering. All fibers exhibited stick-slip friction indicating the successive formation and rupture of strong junctions between the contacting filaments. The static frictional resistance of the sized SiC filament was 4X greater than for the same filament after heat cleaning. This result suggests that the sizing is an organic polymer with a high shear yield strength. Heat cleaning exposes the SiC surface and/or leaves an inorganic residue so that the adhesional contact between filaments has a low fracture energy and frictional sliding occurs by brittle fracture. The frictional resistances of the sized and heat cleaned SiO 2 and glass filaments were all comparable to that of the heat cleaned SiC. It would appear that the sizings as well as the heat cleaned surfaces of the silica and glass have low fracture energies so that the sliding resistance is determined by brittle fracture

  15. Oxidized limonene and oxidized linalool - Concomitant contact allergy to common fragrance terpenes

    DEFF Research Database (Denmark)

    Bråred Christensson, Johanna; Karlberg, Ann Therese; Andersen, Klaus E.

    2016-01-01

    Summary Background Limonene and linalool are common fragrance terpenes. Both oxidized R-limonene and oxidized linalool have recently been patch tested in an international setting, showing contact allergy in 5.2% and 6.9% of dermatitis patients, respectively. Objective To investigate concomitant r...

  16. Needle Terpenes as Chemotaxonomic Markers in Pinus: Subsections Pinus and Pinaster.

    Science.gov (United States)

    Mitić, Zorica S; Jovanović, Snežana Č; Zlatković, Bojan K; Nikolić, Biljana M; Stojanović, Gordana S; Marin, Petar D

    2017-05-01

    Chemical compositions of needle essential oils of 27 taxa from the section Pinus, including 20 and 7 taxa of the subsections Pinus and Pinaster, respectively, were compared in order to determine chemotaxonomic significance of terpenes at infrageneric level. According to analysis of variance, six out of 31 studied terpene characters were characterized by a high level of significance, indicating statistically significant difference between the examined subsections. Agglomerative hierarchical cluster analysis has shown separation of eight groups, where representatives of subsect. Pinaster were distributed within the first seven groups on the dendrogram together with P. nigra subsp. laricio and P. merkusii from the subsect. Pinus. On the other hand, the eighth group included the majority of the members of subsect. Pinus. Our findings, based on terpene characters, complement those obtained from morphological, biochemical, and molecular parameters studied over the past two decades. In addition, results presented in this article confirmed that terpenes are good markers at infrageneric level. © 2017 Wiley-VHCA AG, Zurich, Switzerland.

  17. 75 FR 39450 - Terpene Constituents of the Extract of Chenopodium ambrosioides

    Science.gov (United States)

    2010-07-09

    ..., spices, and other foods and beverages. These three terpene constituents are found naturally in food... ingestion, dermal contact, and inhalation through consumption of foods and beverages, as well as through... toxicity. Acute toxicity studies, submitted to support the registration of the end-use product (EP...

  18. Germacrene A Synthase in Yarrow (Achillea millefolium Is an Enzyme with Mixed Substrate Specificity: Gene Cloning, Functional Characterization and Expression Analysis

    Directory of Open Access Journals (Sweden)

    Leila ePazouki

    2015-03-01

    Full Text Available Terpenoid synthases constitute a highly diverse gene family producing a wide range of cyclic and acyclic molecules consisting of isoprene (C5 residues. Often a single terpene synthase produces a spectrum of molecules of given chain length, but some terpene synthases can use multiple substrates, producing products of different chain length. Only a few such enzymes has been characterized, but the capacity for multiple-substrate use can be more widespread than previously thought. Here we focused on germacrene A synthase (GAS that is a key cytosolic enzyme in the sesquiterpene lactone biosynthesis pathway in the important medicinal plant Achillea millefolium (AmGAS. The full length encoding gene was heterologously expressed in Escherichia coli BL21 (DE3, functionally characterized, and its in vivo expression was analyzed. The recombinant protein catalyzed formation of germacrene A with the C15 substrate farnesyl diphosphate (FDP, while acyclic monoterpenes were formed with the C10 substrate geranyl diphosphate (GDP and cyclic monoterpenes with the C10 substrate neryl diphosphate (NDP. Although monoterpene synthesis has been assumed to be confined exclusively to plastids, AmGAS can potentially synthesize monoterpenes in cytosol when GDP or NDP become available. AmGAS enzyme had high homology with GAS sequences from other Asteraceae species, suggesting that multi-substrate use can be more widespread among germacrene A synthases than previously thought. Expression studies indicated that AmGAS was expressed in both autotrophic and heterotrophic plant compartments with the highest expression levels in leaves and flowers. To our knowledge, this is the first report on the cloning and characterization of germacrene A synthase coding gene in A. millefolium, and multi-substrate use of GAS enzymes.

  19. Virus-induced Gene Silencing-based Functional Analyses Revealed the Involvement of Several Putative Trehalose-6-Phosphate Synthase/Phosphatase Genes in Disease Resistance against Botrytis cinerea and Pseudomonas syringae pv. tomato DC3000 in Tomato

    Directory of Open Access Journals (Sweden)

    Huijuan Zhang

    2016-08-01

    Full Text Available Trehalose and its metabolism have been demonstrated to play important roles in control of plant growth, development and stress responses. However, direct genetic evidence supporting the functions of trehalose and its metabolism in defense response against pathogens is lacking. In the present study, genome-wide characterization of putative trehalose-related genes identified 11 SlTPSs for trehalose-6-phosphate synthase, 8 SlTPPs for trehalose-6-phosphate phosphatase and one SlTRE1 for trehalase in tomato genome. Nine SlTPSs, 4 SlTPPs and SlTRE1 were selected for functional analyses to explore their involvement in tomato disease resistance. Some selected SlTPSs, SlTPPs and SlTRE1 responded with distinct expression induction patterns to Botrytis cinerea and Pseudomonas syringae pv. tomato (Pst DC3000 as well as to defense signaling hormones (e.g. salicylic acid, jasmonic acid and a precursor of ethylene. Virus-induced gene silencing-mediated silencing of SlTPS3, SlTPS4 or SlTPS7 led to deregulation of ROS accumulation and attenuated the expression of defense-related genes upon pathogen infection and thus deteriorated the resistance against B. cinerea or Pst DC3000. By contrast, silencing of SlTPS5 or SlTPP2 led to an increased expression of the defense-related genes upon pathogen infection and conferred an increased resistance against Pst DC3000. Silencing of SlTPS3, SlTPS4, SlTPS5, SlTPS7 or SlTPP2 affected trehalose level in tomato plants with or without infection of B. cinerea or Pst DC3000. These results demonstrate that SlTPS3, SlTPS4, SlTPS5, SlTPS7 and SlTPP2 play roles in resistance against B. cinerea and Pst DC3000, implying the importance of trehalose and tis metabolism in regulation of defense response against pathogens in tomato.

  20. Automated Design and Analysis Tool for CEV Structural and TPS Components, Phase I

    Data.gov (United States)

    National Aeronautics and Space Administration — The innovation of the proposed effort is a unique automated process for the analysis, design, and sizing of CEV structures and TPS. This developed process will...

  1. [Development of a Software for Automatically Generated Contours in Eclipse TPS].

    Science.gov (United States)

    Xie, Zhao; Hu, Jinyou; Zou, Lian; Zhang, Weisha; Zou, Yuxin; Luo, Kelin; Liu, Xiangxiang; Yu, Luxin

    2015-03-01

    The automatic generation of planning targets and auxiliary contours have achieved in Eclipse TPS 11.0. The scripting language autohotkey was used to develop a software for automatically generated contours in Eclipse TPS. This software is named Contour Auto Margin (CAM), which is composed of operational functions of contours, script generated visualization and script file operations. RESULTS Ten cases in different cancers have separately selected, in Eclipse TPS 11.0 scripts generated by the software could not only automatically generate contours but also do contour post-processing. For different cancers, there was no difference between automatically generated contours and manually created contours. The CAM is a user-friendly and powerful software, and can automatically generated contours fast in Eclipse TPS 11.0. With the help of CAM, it greatly save plan preparation time and improve working efficiency of radiation therapy physicists.

  2. USAF TPS L-23 Shear Wind Observed Optimized Path Investigation for NASA (SENIOR ShWOOPIN)

    National Research Council Canada - National Science Library

    Gordon, Randy; Fails, Robert; Baase, Solomon; Eckberg, Jason; Ryan, Charles; Smith, Chris

    2006-01-01

    The SENIOR ShWOOPIN TMP was conducted at the request of the USAF TPS as part of a NASA investigation into the viability of aircraft endurance enhancement through the extraction of energy from horizontal wind gradients...

  3. Investigating the Thermochemical Response of Avcoat TPS from First Principles for Comparison with EFT-1 Data

    Data.gov (United States)

    National Aeronautics and Space Administration — The objective of our work is to develop improved thermal response models of the AVCOAT thermal protection system (TPS) from first principles, and to validate the...

  4. Compartmentation of mono- and sesqui-terpene biosynthesis of the essential oil in poncirus trifoliata

    International Nuclear Information System (INIS)

    Heinrich, G.; Wegener, R.; Schultze, W.

    1980-01-01

    The fruit of Poncirus trifoliata shows glandular cells complexes in the exocarp, which produce a volatile oil rich in monoterpenes but poor in sesquiterpenes and oxigenated compounds. The juice vesicles of the endocarp possess similar cell complexes mainly containing sesquiterpenes and oxigenated compounds, whereas monoterpenes only occur in small amounts. By the use of combined gas chromatography-mass spectrometry 19 components of the rind oil and 15 compounds of the endocarp oil could be identified. As demonstrated by electron microscopy the terpenes most probably are synthesized predominantly, if not exclusively in plastids. As shown by gasradiochromatography radioactive precursors ( 14 Co 2 and 14 C-leucine) are incorporated into mono- and sesqui-terpenes to a different extent. This is due to two gland types producing essential oils of different composition with regard to their mono- and sesqui-terpene percentage. In fruit development the exocarp glands differentiate earlier than the endocarp glands do. The activity of exogenously applied 14 Co 2 first reaches the peripheral glands and later on appears in the interior glands. Depending upon the growth season, labelled leucine transported by the conducting tissues from lower plant parts leads to a high specific activity of the sesqui-terpenes and oxigenated compounds. It could be argued that in this instance the glands of the pulp are better provided with precursors than the exocarp glands. The successive maxima of essential oil production in both glandular complexes, and the changes in the concentration of individual oil constituents during the ontogeny of the fruit also contribute to different incorporation ratios of radioactive precursors into mono- and sesqui-terpenes. (author)

  5. Thin-plate spline (TPS) graphical analysis of the mandible on cephalometric radiographs.

    Science.gov (United States)

    Chang, H P; Liu, P H; Chang, H F; Chang, C H

    2002-03-01

    We describe two cases of Class III malocclusion with and without orthodontic treatment. A thin-plate spline (TPS) analysis of lateral cephalometric radiographs was used to visualize transformations of the mandible. The actual sites of mandibular skeletal change are not detectable with conventional cephalometric analysis. These case analyses indicate that specific patterns of mandibular transformation are associated with Class III malocclusion with or without orthopaedic therapy, and visualization of these deformations is feasible using TPS graphical analysis.

  6. Ceramic Matrix Composite (CMC) Thermal Protection Systems (TPS) and Hot Structures for Hypersonic Vehicles

    Science.gov (United States)

    Glass, David E.

    2008-01-01

    Thermal protection systems (TPS) and hot structures are required for a range of hypersonic vehicles ranging from ballistic reentry to hypersonic cruise vehicles, both within Earth's atmosphere and non-Earth atmospheres. The focus of this paper is on air breathing hypersonic vehicles in the Earth's atmosphere. This includes single-stage to orbit (SSTO), two-stage to orbit (TSTO) accelerators, access to space vehicles, and hypersonic cruise vehicles. This paper will start out with a brief discussion of aerodynamic heating and thermal management techniques to address the high heating, followed by an overview of TPS for rocket-launched and air-breathing vehicles. The argument is presented that as we move from rocket-based vehicles to air-breathing vehicles, we need to move away from the insulated airplane approach used on the Space Shuttle Orbiter to a wide range of TPS and hot structure approaches. The primary portion of the paper will discuss issues and design options for CMC TPS and hot structure components, including leading edges, acreage TPS, and control surfaces. The current state-of-the-art will be briefly discussed for some of the components. The two primary technical challenges impacting the use of CMC TPS and hot structures for hypersonic vehicles are environmental durability and fabrication, and will be discussed briefly.

  7. PENINGKATAN MOTIVASI DAN HASIL BELAJAR MELALUI THINK PAIR SHARE (TPS DI SEKOLAH DASAR

    Directory of Open Access Journals (Sweden)

    Erika Puspitasari

    2016-07-01

    Full Text Available This study aims to improve motivation and learning outcomes through the Think Pair Share (TPS. Subjects were students of class IV SDN Margomulyo 02 Blitar. Type of research is classroom action research. Collecting data using questionnaires and test sheets. Data were analyzed using descriptive analysis. Results penilitian namely (1 there is an increased motivation to learn through TPS with an average score of the first cycle to the second cycle of 18%. (2 there is an increase learning outcomes through TPS with an average score of the first cycle to the second cycle by 14%. Advice given more time guiding students through group discussion so that students can actively work together in groups. Penelitian ini bertujuan untuk meningkatan motivasi dan hasil belajar melalui Think Pair Share (TPS. Subyek penelitian adalah siswa kelas IV SDN Margomulyo 02 Blitar. Jenis penelitiannya adalah penelitian tindakan kelas. Pengumpulan data menggunakan lembar angket dan tes. Teknik analisis data menggunakan analisis deskriptif. Hasil penilitian yaitu (1 terdapat peningkatan motivasi belajar melalui TPS dengan skor rata-rata siklus I ke siklus II sebesar 18%. (2 terdapat peningkatan hasil belajar melalui TPS dengan skor rata-rata siklus I ke siklus II sebesar 14%. Saran yang diberikan lebih membimbing siswa saat diskusi kelompok agar siswa secara aktif dapat bekerja sama dalam kelompok.

  8. Identification and Functional Characterization of Monofunctional ent-Copalyl Diphosphate and ent-Kaurene Synthases in White Spruce Reveal Different Patterns for Diterpene Synthase Evolution for Primary and Secondary Metabolism in Gymnosperms1[W][OA

    Science.gov (United States)

    Keeling, Christopher I.; Dullat, Harpreet K.; Yuen, Mack; Ralph, Steven G.; Jancsik, Sharon; Bohlmann, Jörg

    2010-01-01

    The biosynthesis of the tetracyclic diterpene ent-kaurene is a critical step in the general (primary) metabolism of gibberellin hormones. ent-Kaurene is formed by a two-step cyclization of geranylgeranyl diphosphate via the intermediate ent-copalyl diphosphate. In a lower land plant, the moss Physcomitrella patens, a single bifunctional diterpene synthase (diTPS) catalyzes both steps. In contrast, in angiosperms, the two consecutive cyclizations are catalyzed by two distinct monofunctional enzymes, ent-copalyl diphosphate synthase (CPS) and ent-kaurene synthase (KS). The enzyme, or enzymes, responsible for ent-kaurene biosynthesis in gymnosperms has been elusive. However, several bifunctional diTPS of specialized (secondary) metabolism have previously been characterized in gymnosperms, and all known diTPSs for resin acid biosynthesis in conifers are bifunctional. To further understand the evolution of ent-kaurene biosynthesis as well as the evolution of general and specialized diterpenoid metabolisms in gymnosperms, we set out to determine whether conifers use a single bifunctional diTPS or two monofunctional diTPSs in the ent-kaurene pathway. Using a combination of expressed sequence tag, full-length cDNA, genomic DNA, and targeted bacterial artificial chromosome sequencing, we identified two candidate CPS and KS genes from white spruce (Picea glauca) and their orthologs in Sitka spruce (Picea sitchensis). Functional characterization of the recombinant enzymes established that ent-kaurene biosynthesis in white spruce is catalyzed by two monofunctional diTPSs, PgCPS and PgKS. Comparative analysis of gene structures and enzyme functions highlights the molecular evolution of these diTPSs as conserved between gymnosperms and angiosperms. In contrast, diTPSs for specialized metabolism have evolved differently in angiosperms and gymnosperms. PMID:20044448

  9. Characterization of CYP264B1 and a terpene cyclase of a terpene biosynthesis gene cluster from the myxobacterium Sorangium cellulosum So ce56

    OpenAIRE

    Ly, Thuy Thi Bich

    2011-01-01

    In the work presented here, CYP264B1 and the terpene cyclase GeoA of Sorangium cellulosum So ce56 have been characterized. CYP264B1 is able to convert norisoprenoids (a-ionone and b-ionone) and diverse sesquiterpene compounds, including nootkatone. Three products, 3-hydroxy-a-ionone, 3-hydroxy-b-ionone and 13-hydroxy-nootkatone were characterized using HPLC and 1H and 13C NMR. CYP264B1 is the first enzyme reported to be capable to hydroxylate regioselectively both norisoprenoids at the positi...

  10. Identification and characterization of the iridoid synthase involved in oleuropein biosynthesis in olive (Olea europaea) fruits

    DEFF Research Database (Denmark)

    Alagna, Fiammetta; Geu-Flores, Fernando; Kries, Hajo

    2016-01-01

    The secoiridoids are the main class of specialized metabolites present in olive (Olea europaea L.) fruit. In particular, the secoiridoid oleuropein strongly influences olive oil quality because of its bitterness, which is a desirable trait. In addition, oleuropein possesses a wide range...... of pharmacological properties, including antioxidant, anti-inflammatory, and anti-cancer activities. In accordance, obtaining high oleuropein varieties is a main goal of molecular breeding programs. Here we use a transcriptomic approach to identify candidate genes belonging to the secoiridoid pathway in olive. From...... these candidates, we have functionally characterized the olive homologue of iridoid synthase (OeISY), an unusual terpene cyclase that couples an NAD (P)H-dependent 1,4-reduction step with a subsequent cyclization, and we provide evidence that OeISY likely generates the monoterpene scaffold of oleuropein in olive...

  11. A phycocyanin·phellandrene synthase fusion enhances recombinant protein expression and β-phellandrene (monoterpene) hydrocarbons production in Synechocystis (cyanobacteria).

    Science.gov (United States)

    Formighieri, Cinzia; Melis, Anastasios

    2015-11-01

    Cyanobacteria can be exploited as photosynthetic platforms for heterologous generation of terpene hydrocarbons with industrial applications. Transformation of Synechocystis and heterologous expression of the β-phellandrene synthase (PHLS) gene alone is necessary and sufficient to confer to Synechocystis the ability to divert intermediate terpenoid metabolites and to generate the monoterpene β-phellandrene during photosynthesis. However, terpene synthases, including the PHLS, have a slow Kcat (low Vmax) necessitating high levels of enzyme concentration to enable meaningful rates and yield of product formation. Here, a novel approach was applied to increase the PHLS protein expression alleviating limitations in the rate and yield of β-phellandrene product generation. Different PHLS fusion constructs were generated with the Synechocystis endogenous cpcB sequence, encoding for the abundant in cyanobacteria phycocyanin β-subunit, expressed under the native cpc operon promoter. In one of these constructs, the CpcB·PHLS fusion protein accumulated to levels approaching 20% of the total cellular protein, i.e., substantially higher than expressing the PHLS protein alone under the same endogenous cpc promoter. The CpcB·PHLS fusion protein retained the activity of the PHLS enzyme and catalyzed β-phellandrene synthesis, yielding an average of 3.2 mg product g(-1) dry cell weight (dcw) versus the 0.03 mg g(-1)dcw measured with low-expressing constructs, i.e., a 100-fold yield improvement. In conclusion, the terpene synthase fusion-protein approach is promising, as, in this case, it substantially increased the amount of the PHLS in cyanobacteria, and commensurately improved rates and yield of β-phellandrene hydrocarbons production in these photosynthetic microorganisms. Copyright © 2015 International Metabolic Engineering Society. Published by Elsevier Inc. All rights reserved.

  12. Allylic chlorination of terpenic olefins using a combination of MoCl{sub 5} and NaOCl

    Energy Technology Data Exchange (ETDEWEB)

    Boualy, Brahim; Firdoussi, Larbi El; Ali, Mustapha Ait; Karim, Abdellah, E-mail: elfirdoussi@ucam.ac.m [Universite Cadi Ayyad, Marrakech (Morocco). Faculte des Sciences Semlalia. Lab. de Chimie de Coordination

    2011-07-01

    MoCl{sub 5} is applied as efficient agent in allylic chlorination of terpenic olefins in the presence of NaOCl as chlorine donor. Various terpenes are converted to the corresponding allylic chlorides in moderate to good yield under mild and optimized reaction conditions. Different molybdenum precursors are also studied. Among them, MoO{sub 3} gives good yield, but after a longer reaction time. (author)

  13. Effects of phosphorus availability and genetic variation of leaf terpene content and emission rate in Pinus pinaster seedlings susceptible and resistant to the pine weevil, Hylobius abietis.

    Science.gov (United States)

    Blanch, J-S; Sampedro, L; Llusià, J; Moreira, X; Zas, R; Peñuelas, J

    2012-03-01

    We studied the effects of phosphorus fertilisation on foliar terpene concentrations and foliar volatile terpene emission rates in six half-sib families of Pinus pinaster Ait. seedlings. Half of the seedlings were resistant to attack of the pine weevil Hylobius abietis L., a generalist phloem feeder, and the remaining seedlings were susceptible to this insect. We hypothesised that P stress could modify the terpene concentration in the needles and thus lead to altered terpene emission patterns relevant to plant-insect signalling. The total concentration and emission rate ranged between 5732 and 13,995 μg·g(-1) DW and between 2 and 22 μg·g(-1) DW·h(-1), respectively. Storage and emission were dominated by the isomers α- and β-pinene (77.2% and 84.2% of the total terpene amount amassed and released, respectively). In both resistant and susceptible families, P stress caused an increase of 31% in foliar terpene concentration with an associated 5-fold decrease in terpene emission rates. A higher terpene content in the leaves implies that the 'excess carbon', available under limiting growth conditions (P scarcity), is allocated to terpene production. Sensitive families showed a greater increase in terpene emission rates with increasing P concentrations, which could explain their susceptibility to H. abietis. © 2011 German Botanical Society and The Royal Botanical Society of the Netherlands.

  14. Fungal endophytes – the hidden inducers of volatile terpene biosynthesis in tomato plants

    DEFF Research Database (Denmark)

    Ntana, Fani; Jensen, Birgit; Jørgensen, Hans Jørgen Lyngs

    mycorrhizal spores in the Indian Thar desert, colonizes the root cortex of a wide range of plants, enhancing plant growth and modulating plant specialized metabolism. The effect of S. indica colonization on the metabolism of the host can be potentially used in improving plant defence against pathogens...... and herbivores. Tomato (Solanum lycopersicum) is an important crop, often challenged by fungal pathogens and insect pests. The wide variety of secondary metabolites produced by the plant, and especially terpenes, play a crucial role in plant defence, helping in repelling possible enemies. This project is focused....... indica-inoculated and S. indica-free tomato plants. Preliminary data suggest that fungal colonization results in increased production of specific volatile terpenes. A transcriptome analysis on fungus-associated and fungus-free plant tissues is currently ongoing to elucidate in depth the mechanisms...

  15. [Regulation of terpene metabolism]. Annual progress report, March 15, 1989--March 14, 1990

    Energy Technology Data Exchange (ETDEWEB)

    Croteau, R.

    1989-11-09

    Terpenoid oils, resins, and waxes from plants are important renewable resources. The objective of this project is to understand the regulation of terpenoid metabolism using the monoterpenes (C{sub 10}) as a model. The pathways of monoterpene biosynthesis and catabolism have been established, and the relevant enzymes characterized. Developmental studies relating enzyme levels to terpene accumulation within the oil gland sites of synthesis, and work with bioregulators, indicate that monoterpene production is controlled by terpene cyclases, the enzymes catalyzing the first step of the monoterpene pathway. As the leaf oil glands mature, cyclase levels decline and monoterpene biosynthesis ceases. Yield then decreases as the monoterpenes undergo catabolism by a process involving conversion to a glycoside and transport from the leaf glands to the root. At this site, the terpenoid is oxidatively degraded to acetate that is recycled into other lipid metabolites. During the transition from terpene biosynthesis to catabolism, the oil glands undergo dramatic ultrastructural modification. Degradation of the producing cells results in mixing of previously compartmentized monoterpenes with the catabolic enzymes, ultimately leading to yield decline. This regulatory model is being applied to the formation of other terpenoid classes (C{sub 15} C{sub 20}, C{sub 30}, C{sub 40}) within the oil glands. Preliminary investigations on the formation of sesquiterpenes (C{sub 15}) suggest that the corresponding cyclases may play a lesser role in determining yield of these products, but that compartmentation effects are important. From these studies, a comprehensive scheme for the regulation of terpene metabolism is being constructed. Results from this project wail have important consequences for the yield and composition of terpenoid natural products that can be made available for industrial exploitation.

  16. CYP109E1 is a novel versatile statin and terpene oxidase from Bacillus megaterium.

    Science.gov (United States)

    Putkaradze, Natalia; Litzenburger, Martin; Abdulmughni, Ammar; Milhim, Mohammed; Brill, Elisa; Hannemann, Frank; Bernhardt, Rita

    2017-12-01

    CYP109E1 is a cytochrome P450 monooxygenase from Bacillus megaterium with a hydroxylation activity for testosterone and vitamin D3. This study reports the screening of a focused library of statins, terpene-derived and steroidal compounds to explore the substrate spectrum of this enzyme. Catalytic activity of CYP109E1 towards the statin drug-precursor compactin and the prodrugs lovastatin and simvastatin as well as biotechnologically relevant terpene compounds including ionones, nootkatone, isolongifolen-9-one, damascones, and β-damascenone was found in vitro. The novel substrates induced a type I spin-shift upon binding to P450 and thus permitted to determine dissociation constants. For the identification of conversion products by NMR spectroscopy, a B. megaterium whole-cell system was applied. NMR analysis revealed for the first time the ability of CYP109E1 to catalyze an industrially highly important reaction, the production of pravastatin from compactin, as well as regioselective oxidations generating drug metabolites (6'β-hydroxy-lovastatin, 3'α-hydroxy-simvastatin, and 4″-hydroxy-simvastatin) and valuable terpene derivatives (3-hydroxy-α-ionone, 4-hydroxy-β-ionone, 11,12-epoxy-nootkatone, 4(R)-hydroxy-isolongifolen-9-one, 3-hydroxy-α-damascone, 4-hydroxy-β-damascone, and 3,4-epoxy-β-damascone). Besides that, a novel compound, 2-hydroxy-β-damascenone, produced by CYP109E1 was identified. Docking calculations using the crystal structure of CYP109E1 rationalized the experimentally observed regioselective hydroxylation and identified important amino acid residues for statin and terpene binding.

  17. STRATEGI PENENTUAN LOKASI DAN KEBUTUHAN LAHAN TPS (TEMPAT PENAMPUNGAN SEMENTARA SAMPAH BERDASARKAN FUNGSI KAWASAN DI KOTA DENPASAR

    Directory of Open Access Journals (Sweden)

    Ismid Achmad

    2015-08-01

    Full Text Available The percentage of solid waste services Denpasar City in 2013 reached 51,8 percent, the limited number and capacity TPS in Denpasar not meet the needs of waste management services in denpasar. The study was to examine the relevant strategies appropriate to the criteria and requirements related to the determination of the location of TPS and the land need. The position of the study is Part Of The City (BWK Central in Denpasar. Analysis of technical aspects will evaluation garbage production in order to know the ideal needs of TPS, the evaluation of TPS placement based on the function of the area and the land needed. Environmental aspect will analyze the environmental risk due to the presence of a TPS. Analysis of public opinion and the behavior of people will use a questionnaire in the collection of data to be analyzed with qualitative methods as a consideration in determining the location and design of a TPS. Institutional aspect will be evaluated about the understanding DKP (Office of Cleaning and Gardening Services employees of the City of Denpasar in the management of garbage, especially in the TPS. As for determining the direction and strategy of the management of garbage SWOT analysis was used.Based on the analysis, the need for the addition of TPS to improve the capacity of TPS, the shape of TPS had comply standard of SNI and environmental safety factor. Additions and changes in the shape of TPS also effect land requirement, the total area of land that must be reserved for TPS is 0,40 hectares. Determining the location of TPS is based on the need of the type of TPS over the spatial pattern. Analysis of public opinion on the determination of the location of TPS, it is obtained that the distance of a TPS with the source must be relatively close and the land use is a vacant land. It is also suggested optimizing the role of the DKP of Denpasar by increasing socialization and community development programs in waste management.

  18. SUMO-fusion, purification, and characterization of a (+)-zizaene synthase from Chrysopogon zizanioides

    International Nuclear Information System (INIS)

    Hartwig, S.; Frister, T.; Alemdar, S.; Li, Z.; Scheper, T.; Beutel, S.

    2015-01-01

    An uncharacterized plant cDNA coding for a polypeptide presumably having sesquiterpene synthase activity, was expressed in soluble and active form. Two expression strategies were evaluated in Escherichia coli. The enzyme was fused to a highly soluble SUMO domain, in addition to being produced in an unfused form by a cold-shock expression system. Yields up to ∼325 mg/L −1 were achieved in batch cultivations. The 6x-His-tagged enzyme was purified employing an Ni 2+ -IMAC-based procedure. Identity of the protein was established by Western Blot analysis as well as peptide mass fingerprinting. A molecular mass of 64 kDa and an isoelectric point of pI 4.95 were determined by 2D gel electrophoresis. Cleavage of the fusion domain was possible by digestion with specific SUMO protease. The synthase was active in Mg 2+ containing buffer and catalyzed the production of (+)-zizaene (syn. khusimene), a precursor of khusimol, from farnesyl diphosphate. Product identity was confirmed by GC–MS and comparison of retention indices. Enzyme kinetics were determined by measuring initial reaction rates for the product, using varying substrate concentrations. By assuming a Michaelis–Menten model, kinetic parameters of K M  = 1.111 μM (±0.113), v max  = 0.3245 μM min −1 (±0.0035), k cat  = 2.95 min −1 , as well as a catalytic efficiency k cat /K M  = 4.43 × 10 4  M −1 s −1 were calculated. Fusion to a SUMO moiety can substantially increase soluble expression levels of certain hard to express terpene synthases in E. coli. The kinetic data determined for the recombinant synthase are comparable to other described plant sesquiterpene synthases and in the typical range of enzymes belonging to the secondary metabolism. This leaves potential for optimizing catalytic parameters through methods like directed evolution. - Highlights: • Uncharacterized (+)-zizaene synthase from C. zizanoides was cloned and expressed. • Fusion to SUMO and cold-shock induction

  19. SUMO-fusion, purification, and characterization of a (+)-zizaene synthase from Chrysopogon zizanioides

    Energy Technology Data Exchange (ETDEWEB)

    Hartwig, S.; Frister, T.; Alemdar, S.; Li, Z.; Scheper, T.; Beutel, S., E-mail: beutel@iftc.uni-hannover.de

    2015-03-20

    An uncharacterized plant cDNA coding for a polypeptide presumably having sesquiterpene synthase activity, was expressed in soluble and active form. Two expression strategies were evaluated in Escherichia coli. The enzyme was fused to a highly soluble SUMO domain, in addition to being produced in an unfused form by a cold-shock expression system. Yields up to ∼325 mg/L{sup −1} were achieved in batch cultivations. The 6x-His-tagged enzyme was purified employing an Ni{sup 2+}-IMAC-based procedure. Identity of the protein was established by Western Blot analysis as well as peptide mass fingerprinting. A molecular mass of 64 kDa and an isoelectric point of pI 4.95 were determined by 2D gel electrophoresis. Cleavage of the fusion domain was possible by digestion with specific SUMO protease. The synthase was active in Mg{sup 2+} containing buffer and catalyzed the production of (+)-zizaene (syn. khusimene), a precursor of khusimol, from farnesyl diphosphate. Product identity was confirmed by GC–MS and comparison of retention indices. Enzyme kinetics were determined by measuring initial reaction rates for the product, using varying substrate concentrations. By assuming a Michaelis–Menten model, kinetic parameters of K{sub M} = 1.111 μM (±0.113), v{sub max} = 0.3245 μM min{sup −1} (±0.0035), k{sub cat} = 2.95 min{sup −1}, as well as a catalytic efficiency k{sub cat}/K{sub M} = 4.43 × 10{sup 4} M{sup −1} s{sup −1} were calculated. Fusion to a SUMO moiety can substantially increase soluble expression levels of certain hard to express terpene synthases in E. coli. The kinetic data determined for the recombinant synthase are comparable to other described plant sesquiterpene synthases and in the typical range of enzymes belonging to the secondary metabolism. This leaves potential for optimizing catalytic parameters through methods like directed evolution. - Highlights: • Uncharacterized (+)-zizaene synthase from C. zizanoides was cloned

  20. Glycogen Metabolic Genes Are Involved in Trehalose-6-Phosphate Synthase-Mediated Regulation of Pathogenicity by the Rice Blast Fungus Magnaporthe oryzae

    Science.gov (United States)

    Wilson, Richard A.; Wang, Zheng-Yi; Kershaw, Michael J.; Talbot, Nicholas J.

    2013-01-01

    The filamentous fungus Magnaporthe oryzae is the causal agent of rice blast disease. Here we show that glycogen metabolic genes play an important role in plant infection by M. oryzae. Targeted deletion of AGL1 and GPH1, which encode amyloglucosidase and glycogen phosphorylase, respectively, prevented mobilisation of glycogen stores during appressorium development and caused a significant reduction in the ability of M. oryzae to cause rice blast disease. By contrast, targeted mutation of GSN1, which encodes glycogen synthase, significantly reduced the synthesis of intracellular glycogen, but had no effect on fungal pathogenicity. We found that loss of AGL1 and GPH1 led to a reduction in expression of TPS1 and TPS3, which encode components of the trehalose-6-phosphate synthase complex, that acts as a genetic switch in M. oryzae. Tps1 responds to glucose-6-phosphate levels and the balance of NADP/NADPH to regulate virulence-associated gene expression, in association with Nmr transcriptional inhibitors. We show that deletion of the NMR3 transcriptional inhibitor gene partially restores virulence to a Δagl1Δgph1 mutant, suggesting that glycogen metabolic genes are necessary for operation of the NADPH-dependent genetic switch in M. oryzae. PMID:24098112

  1. Accuracy Evaluation of Oncentra™ TPS in HDR Brachytherapy of Nasopharynx Cancer Using EGSnrc Monte Carlo Code

    Science.gov (United States)

    Hadad, K.; Zohrevand, M.; Faghihi, R.; Sedighi Pashaki, A.

    2015-01-01

    Background HDR brachytherapy is one of the commonest methods of nasopharyngeal cancer treatment. In this method, depending on how advanced one tumor is, 2 to 6 Gy dose as intracavitary brachytherapy is prescribed. Due to high dose rate and tumor location, accuracy evaluation of treatment planning system (TPS) is particularly important. Common methods used in TPS dosimetry are based on computations in a homogeneous phantom. Heterogeneous phantoms, especially patient-specific voxel phantoms can increase dosimetric accuracy. Materials and Methods In this study, using CT images taken from a patient and ctcreate-which is a part of the DOSXYZnrc computational code, patient-specific phantom was made. Dose distribution was plotted by DOSXYZnrc and compared with TPS one. Also, by extracting the voxels absorbed dose in treatment volume, dose-volume histograms (DVH) was plotted and compared with Oncentra™ TPS DVHs. Results The results from calculations were compared with data from Oncentra™ treatment planning system and it was observed that TPS calculation predicts lower dose in areas near the source, and higher dose in areas far from the source relative to MC code. Absorbed dose values in the voxels also showed that TPS reports D90 value is 40% higher than the Monte Carlo method. Conclusion Today, most treatment planning systems use TG-43 protocol. This protocol may results in errors such as neglecting tissue heterogeneity, scattered radiation as well as applicator attenuation. Due to these errors, AAPM emphasized departing from TG-43 protocol and approaching new brachytherapy protocol TG-186 in which patient-specific phantom is used and heterogeneities are affected in dosimetry. PMID:25973408

  2. Conditioning of the vacuum system of the TPS storage ring without baking in situ

    Energy Technology Data Exchange (ETDEWEB)

    Chan, C.K., E-mail: ckchan@nsrrc.org.tw; Chang, C.C.; Shueh, C.; Yang, I.C.; Wu, L.H.; Chen, B.Y.; Cheng, C.M.; Huang, Y.T.; Chuang, J.Y.; Cheng, Y.T.; Hsiao, Y.M.; Sheng, Albert

    2017-04-11

    To shorten the machine downtime, a maintenance procedure without baking in situ has been developed and applied to maintain and to upgrade the vacuum system of the TPS storage ring. The data of photon-stimulated desorption (PSD) reveal no obvious discrepancy between baking and not baking the vacuum system in situ. A beam-conditioning dose of extent only 11.8 A h is required to recover quickly the dynamic pressure of an unbaked vacuum system to its pre-intervention value according to the TPS maintenance experience.

  3. Conditioning of the vacuum system of the TPS storage ring without baking in situ

    Science.gov (United States)

    Chan, C. K.; Chang, C. C.; Shueh, C.; Yang, I. C.; Wu, L. H.; Chen, B. Y.; Cheng, C. M.; Huang, Y. T.; Chuang, J. Y.; Cheng, Y. T.; Hsiao, Y. M.; Sheng, Albert

    2017-04-01

    To shorten the machine downtime, a maintenance procedure without baking in situ has been developed and applied to maintain and to upgrade the vacuum system of the TPS storage ring. The data of photon-stimulated desorption (PSD) reveal no obvious discrepancy between baking and not baking the vacuum system in situ. A beam-conditioning dose of extent only 11.8 A h is required to recover quickly the dynamic pressure of an unbaked vacuum system to its pre-intervention value according to the TPS maintenance experience.

  4. Conditioning of the vacuum system of the TPS storage ring without baking in situ

    International Nuclear Information System (INIS)

    Chan, C.K.; Chang, C.C.; Shueh, C.; Yang, I.C.; Wu, L.H.; Chen, B.Y.; Cheng, C.M.; Huang, Y.T.; Chuang, J.Y.; Cheng, Y.T.; Hsiao, Y.M.; Sheng, Albert

    2017-01-01

    To shorten the machine downtime, a maintenance procedure without baking in situ has been developed and applied to maintain and to upgrade the vacuum system of the TPS storage ring. The data of photon-stimulated desorption (PSD) reveal no obvious discrepancy between baking and not baking the vacuum system in situ. A beam-conditioning dose of extent only 11.8 A h is required to recover quickly the dynamic pressure of an unbaked vacuum system to its pre-intervention value according to the TPS maintenance experience.

  5. Fiber Optic Temperature Sensors in TPS: Arc Jet Model Design & Testing

    Science.gov (United States)

    Black, Richard; Feldman, Jay; Ellerby, Donald; Monk, Joshua; Moslehi, Behzad; Oblea, Levy; Switzer, Matthew

    2017-01-01

    Techniques for using fiber optics with Fiber Bragg Gratings (FBGs) have been developed by IFOS Corp. for use in thermal protection systems (TPS) on spacecraft heat shield materials through NASA Phase 1 and 2 SBIR efforts and have been further improved in a recent collaboration between IFOS and NASA that will be described here. Fiber optic temperature sensors offer several potential advantages over traditional thermocouple sensors including a) multiplexing many sensors in a single fiber to increase sensor density in a given array or to provide spatial resolution, b) improved thermal property match between sensor and TPS to reduce heat flow disruption, c) lack of electrical conductivity.

  6. TPS-HAMMER: improving HAMMER registration algorithm by soft correspondence matching and thin-plate splines based deformation interpolation.

    Science.gov (United States)

    Wu, Guorong; Yap, Pew-Thian; Kim, Minjeong; Shen, Dinggang

    2010-02-01

    We present an improved MR brain image registration algorithm, called TPS-HAMMER, which is based on the concepts of attribute vectors and hierarchical landmark selection scheme proposed in the highly successful HAMMER registration algorithm. We demonstrate that TPS-HAMMER algorithm yields better registration accuracy, robustness, and speed over HAMMER owing to (1) the employment of soft correspondence matching and (2) the utilization of thin-plate splines (TPS) for sparse-to-dense deformation field generation. These two aspects can be integrated into a unified framework to refine the registration iteratively by alternating between soft correspondence matching and dense deformation field estimation. Compared with HAMMER, TPS-HAMMER affords several advantages: (1) unlike the Gaussian propagation mechanism employed in HAMMER, which can be slow and often leaves unreached blotches in the deformation field, the deformation interpolation in the non-landmark points can be obtained immediately with TPS in our algorithm; (2) the smoothness of deformation field is preserved due to the nice properties of TPS; (3) possible misalignments can be alleviated by allowing the matching of the landmarks with a number of possible candidate points and enforcing more exact matches in the final stages of the registration. Extensive experiments have been conducted, using the original HAMMER as a comparison baseline, to validate the merits of TPS-HAMMER. The results show that TPS-HAMMER yields significant improvement in both accuracy and speed, indicating high applicability for the clinical scenario. Copyright (c) 2009 Elsevier Inc. All rights reserved.

  7. Prognostic significance of tissue polypeptidespecific antigen (TPS) in patients with advanced non-small cell lung cancer

    NARCIS (Netherlands)

    A. van der Gaast (Ate); C.H.H. Schoenmakers (Christian); T.C. Kok (Tjebbe); B.G. Blijenberg (Bert); W.C.J. Hop (Wim); T.A.W. Splinter (Ted)

    1994-01-01

    textabstractIn this study, we evaluated the prognostic value of the tumour marker, tissue polypeptide-specific antigen (TPS), in 203 patients with non-small cell lung cancer (NSCLC), and related this to several other known prognostic factors. TPS was significantly correlated with lactate

  8. Structure of the ent-Copalyl Diphosphate Synthase PtmT2 from Streptomyces platensis CB00739, a Bacterial Type II Diterpene Synthase.

    Science.gov (United States)

    Rudolf, Jeffrey D; Dong, Liao-Bin; Cao, Hongnan; Hatzos-Skintges, Catherine; Osipiuk, Jerzy; Endres, Michael; Chang, Chin-Yuan; Ma, Ming; Babnigg, Gyorgy; Joachimiak, Andrzej; Phillips, George N; Shen, Ben

    2016-08-31

    Terpenoids are the largest and most structurally diverse family of natural products found in nature, yet their presence in bacteria is underappreciated. The carbon skeletons of terpenoids are generated through carbocation-dependent cyclization cascades catalyzed by terpene synthases (TSs). Type I and type II TSs initiate cyclization via diphosphate ionization and protonation, respectively, and protein structures of both types are known. Most plant diterpene synthases (DTSs) possess three α-helical domains (αβγ), which are thought to have arisen from the fusion of discrete, ancestral bacterial type I TSs (α) and type II TSs (βγ). Type II DTSs of bacterial origin, of which there are no structurally characterized members, are a missing piece in the structural evolution of TSs. Here, we report the first crystal structure of a type II DTS from bacteria. PtmT2 from Streptomyces platensis CB00739 was verified as an ent-copalyl diphosphate synthase involved in the biosynthesis of platensimycin and platencin. The crystal structure of PtmT2 was solved at a resolution of 1.80 Å, and docking studies suggest the catalytically active conformation of geranylgeranyl diphosphate (GGPP). Site-directed mutagenesis confirmed residues involved in binding the diphosphate moiety of GGPP and identified DxxxxE as a potential Mg(2+)-binding motif for type II DTSs of bacterial origin. Finally, both the shape and physicochemical properties of the active sites are responsible for determining specific catalytic outcomes of TSs. The structure of PtmT2 fundamentally advances the knowledge of bacterial TSs, their mechanisms, and their role in the evolution of TSs.

  9. Evolution of the Cannabinoid and Terpene Content during the Growth of Cannabis sativa Plants from Different Chemotypes.

    Science.gov (United States)

    Aizpurua-Olaizola, Oier; Soydaner, Umut; Öztürk, Ekin; Schibano, Daniele; Simsir, Yilmaz; Navarro, Patricia; Etxebarria, Nestor; Usobiaga, Aresatz

    2016-02-26

    The evolution of major cannabinoids and terpenes during the growth of Cannabis sativa plants was studied. In this work, seven different plants were selected: three each from chemotypes I and III and one from chemotype II. Fifty clones of each mother plant were grown indoors under controlled conditions. Every week, three plants from each variety were cut and dried, and the leaves and flowers were analyzed separately. Eight major cannabinoids were analyzed via HPLC-DAD, and 28 terpenes were quantified using GC-FID and verified via GC-MS. The chemotypes of the plants, as defined by the tetrahydrocannabinolic acid/cannabidiolic acid (THCA/CBDA) ratio, were clear from the beginning and stable during growth. The concentrations of the major cannabinoids and terpenes were determined, and different patterns were found among the chemotypes. In particular, the plants from chemotypes II and III needed more time to reach peak production of THCA, CBDA, and monoterpenes. Differences in the cannabigerolic acid development among the different chemotypes and between monoterpene and sesquiterpene evolution patterns were also observed. Plants of different chemotypes were clearly differentiated by their terpene content, and characteristic terpenes of each chemotype were identified.

  10. The Contents of Terpene Trilactone and Flavonoid in Leaves of Seedlings from Ancient Female Ginkgo Trees in China

    Directory of Open Access Journals (Sweden)

    Yanhui ZHANG

    2017-07-01

    Full Text Available Flavonoids and terpene trilactones, especially, ginkgo flavonglycosides, ginkgolides and bilobalides in leaves of ginkgo trees, need to be studied for effective application of these active components with high medical and health-care values. This study was aimed to provide scientific bases for genealogies selection and harvest season confirmation for Ginkgo biloba. A high-performance liquid chromatographic method (HPLC-ELSD was developed to determine the contents of terpene trilactone and flavonoid of 36 ancient G. biloba genealogies from 19 provinces in China. The study indicated that the content gradually increased from April to August, and thereafter declined. Analysis of variance indicated that the contents of terpene trilactone, flavonoids, and their respective components had significant difference among 36 genealogies. The cluster analysis showed that No. 72 (Xing'an, Guangxi, No. 58 (Youyang, Chongqing, No. 82 (Rugao, Jiangsu, No. 123 (Huixian, Gansu, No. 99 (Dujun, Guizhou, No. 10 (Tai'an, Shandong and No. 133 (Mentougou, Beijing genealogies have higher content of terpene trilactone and flavonoid. These results can help us to select superior variety containing high content of terpene trilactone and flavonoid.

  11. CloudTPS: Scalable Transactions for Web Applications in the Cloud

    NARCIS (Netherlands)

    Zhou, W.; Pierre, G.E.O.; Chi, C.-H.

    2010-01-01

    NoSQL Cloud data services provide scalability and high availability properties for web applications but at the same time they sacrifice data consistency. However, many applications cannot afford any data inconsistency. CloudTPS is a scalable transaction manager to allow cloud database services to

  12. SU-E-J-89: Deformable Registration Method Using B-TPS in Radiotherapy.

    Science.gov (United States)

    Xie, Y

    2012-06-01

    A novel deformable registration method for four-dimensional computed tomography (4DCT) images is developed in radiation therapy. The proposed method combines the thin plate spline (TPS) and B-spline together to achieve high accuracy and high efficiency. The method consists of two steps. First, TPS is used as a global registration method to deform large unfit regions in the moving image to match counterpart in the reference image. Then B-spline is used for local registration, the previous deformed moving image is further deformed to match the reference image more accurately. Two clinical CT image sets, including one pair of lung and one pair of liver, are simulated using the proposed algorithm, which results in a tremendous improvement in both run-time and registration quality, compared with the conventional methods solely using either TPS or B-spline. The proposed method can combine the efficiency of TPS and the accuracy of B-spline, performing good adaptively and robust in registration of clinical 4DCT image. © 2012 American Association of Physicists in Medicine.

  13. Prognostic significance of CA 125 and TPS levels after 3 chemotherapy courses in ovarian cancer patients

    NARCIS (Netherlands)

    van Dalen, A; Favier, J; Burges, A; Hasholzner, U; de Bruijn, HWA; Dobler-Girdziunaite, D; Dombi, VH; Fink, D; Giai, M; McGing, P; Harlozinska, A; Kainz, C; Markowska, J; Molina, R; Sturgeon, C; Bowman, A; Einarsson, R

    2000-01-01

    Objective. To evaluate the prognostic significance of and predictive value for survival of CA 125 and TPS levels after three chemotherapy courses in ovarian cancer patients. Methods. We analyzed in a prospective multicenter study the 1- and 2-year overall survival (OS) in ovarian carcinoma patients.

  14. TPS as an Effective Technique to Enhance the Students' Achievement on Writing Descriptive Text

    Science.gov (United States)

    Sumarsih, M. Pd.; Sanjaya, Dedi

    2013-01-01

    Students' achievement in writing descriptive text is very low, in this study Think Pair Share (TPS) is applied to solve the problem. Action research is conducted for the result. Additionally, qualitative and quantitative techniques are applied in this research. The subject of this research is grade VIII in Junior High School in Indonesia. From…

  15. Decreasing the emittance using a multi-period Robinson wigglers in TPS

    Energy Technology Data Exchange (ETDEWEB)

    Huamg, C. W., E-mail: huang.zw@nsrrc.org.tw [Department of Physics, National Tsing Hua University Hsinchu 30043, Taiwan (China); Hwang, C. S., E-mail: cshwang@nsrrc.org.tw [NSRRC, 101 Hsin-Ann Road, Hsinchu Science Park, Hsinchu 30076, Taiwan (China); Department of Electrophysics, National Chiao Tung University, Hsinchu 300, Taiwan (China); Lee, S. Y., E-mail: shylee@indiana.edu [Department of Physics, Indiana University (United States)

    2016-07-27

    The Taiwan Photon Source (TPS) has been successfully commissioned. However, the minimum emittance in the TPS lattice is 1.6 nm rad. In the existing TPS storage ring lattice, it is imperative to reduce the emittance to below 1 nm rad. Therefore, a feasibility study for reducing the effective emittance of the TPS storage ring by using a Robinson wiggler was launched; the reduction is necessary to enhance the photon brilliance. In this study, a permanent-magnet multiperiod Robinson wiggler (MRW) was developed for use instead of the single-period Robinson wiggler. In general, the quadruple field of a combined function magnet in the storage ring is approximately few tesla per meter. According to beam dynamic analysis, we found that it is necessary to adopt a high gradient (40 T/m) combined-function MRW magnet to reduce the emittance effectively. Therefore, a high gradient field strength is required in the combined function MRW magnet. In this study, the quadrupole field strength of the MRW magnet was allowed to be approximately 40 T/m at a magnet gap of 20 mm. The period length of the MRW magnet was 300 mm and the period number was 16. The of MRWs is discussed in regard to the possibility of increasing the photon brilliance from IU22.

  16. PEMBELAJARAN READING-CONCEPT-MAP THINK PAIR SHARE (REMAP TPS DAPAT MENINGKATKAN HASIL BELAJAR KOGNITIF

    Directory of Open Access Journals (Sweden)

    Miswandi Tendrita

    2017-06-01

    Full Text Available The aims of this study are to explore the effect of Reading-Concept Map- Think Pair Share (Remap-TPS towards cognitive learning outcomes on highschool students. The participant for the study consisted of 54 students from the Senior High School 1, Batu City. This research used Nonequivalent Pretest-Posttest Control Group Design.  Cognitive learning outcome obtained from the essay test. The test results were then analyzed with Anacova. The results of this experiment showed that Remap-TPS can enhance cognitive learning outcomes. Penelitian ini bertujuan untuk mengetahui pengaruh pembelajaran Reading-Concept Map-Think Pair Share (Remap-TPS terhadap hasil belajar kognitif siswa. Penelitian ini dilakukan di SMA Negeri 1 Batu dengan jumlah partistipan sebanyak 54 siswa. Rancangan penelitian yang digunakan adalah nonequivalent pretest-posttest control group design. Data hasil belajar kognitif diperoleh dari hasil tes essay, dan hasilnya dianalisis dengan teknik Anacova. Hasil penelitian menunjukkan bahwa Remap-TPS berpengaruh terhadap hasil belajar kognitif siswa.

  17. Functional and Structural Characterization of a (+)-Limonene Synthase from Citrus sinensis.

    Science.gov (United States)

    Morehouse, Benjamin R; Kumar, Ramasamy P; Matos, Jason O; Olsen, Sarah Naomi; Entova, Sonya; Oprian, Daniel D

    2017-03-28

    Terpenes make up the largest and most diverse class of natural compounds and have important commercial and medical applications. Limonene is a cyclic monoterpene (C 10 ) present in nature as two enantiomers, (+) and (-), which are produced by different enzymes. The mechanism of production of the (-)-enantiomer has been studied in great detail, but to understand how enantiomeric selectivity is achieved in this class of enzymes, it is important to develop a thorough biochemical description of enzymes that generate (+)-limonene, as well. Here we report the first cloning and biochemical characterization of a (+)-limonene synthase from navel orange (Citrus sinensis). The enzyme obeys classical Michaelis-Menten kinetics and produces exclusively the (+)-enantiomer. We have determined the crystal structure of the apoprotein in an "open" conformation at 2.3 Å resolution. Comparison with the structure of (-)-limonene synthase (Mentha spicata), which is representative of a fully closed conformation (Protein Data Bank entry 2ONG ), reveals that the short H-α1 helix moves nearly 5 Å inward upon substrate binding, and a conserved Tyr flips to point its hydroxyl group into the active site.

  18. MO-F-16A-01: Implementation of MPPG TPS Verification Tests On Various Accelerators

    Energy Technology Data Exchange (ETDEWEB)

    Smilowitz, J; Bredfeldt, J; Geurts, M; Miller, J [University of Wisconsin, Madison, WI (United States)

    2014-06-15

    Purpose: To demonstrate the implementation of the Medical Physics Practice Guideline (MPPG) for dose calculation and beam parameters verification of treatment planning systems (TPS). Methods: We implemented the draft TPS MPPG for three linacs: Varian Trilogy, TomoHDA and Elekta Infinity. Static and modulated test plans were created. The static fields are different than used in commissioning. Data was collected using ion chambers and diodes in a scanning water tank, Delta4 phantom and a custom phantom. MatLab and Microsoft Excel were used to create analysis tools to compare reference DICOM dose with scan data. This custom code allowed for the interpolation, registration and gamma analysis of arbitrary dose profiles. It will be provided as open source code. IMRT fields were validated with Delta4 registration and comparison tools. The time for each task was recorded. Results: The tests confirmed the strengths, and revealed some limitations, of our TPS. The agreement between calculated and measured dose was reported for all beams. For static fields, percent depth dose and profiles were analyzed with criteria in the draft MPPG. The results reveal areas of slight mismatch with the model (MLC leaf penumbra, buildup region.) For TomoTherapy, the IMRT plan 2%/2 mm gamma analysis revealed poorest agreement in the low dose regions. For one static test plan for all 10MV Trilogy photon beams, the plan generation, scan queue creation, data collection, data analysis and report took 2 hours, excluding tank setup. Conclusions: We have demonstrated the implementation feasibility of the TPS MPPG. This exercise generated an open source tool for dose comparisons between scan data and DICOM dose data. An easily reproducible and efficient infrastructure with streamlined data collection was created for repeatable robust testing of the TPS. The tests revealed minor discrepancies in our models and areas for improvement that are being investigated.

  19. MO-F-16A-01: Implementation of MPPG TPS Verification Tests On Various Accelerators

    International Nuclear Information System (INIS)

    Smilowitz, J; Bredfeldt, J; Geurts, M; Miller, J

    2014-01-01

    Purpose: To demonstrate the implementation of the Medical Physics Practice Guideline (MPPG) for dose calculation and beam parameters verification of treatment planning systems (TPS). Methods: We implemented the draft TPS MPPG for three linacs: Varian Trilogy, TomoHDA and Elekta Infinity. Static and modulated test plans were created. The static fields are different than used in commissioning. Data was collected using ion chambers and diodes in a scanning water tank, Delta4 phantom and a custom phantom. MatLab and Microsoft Excel were used to create analysis tools to compare reference DICOM dose with scan data. This custom code allowed for the interpolation, registration and gamma analysis of arbitrary dose profiles. It will be provided as open source code. IMRT fields were validated with Delta4 registration and comparison tools. The time for each task was recorded. Results: The tests confirmed the strengths, and revealed some limitations, of our TPS. The agreement between calculated and measured dose was reported for all beams. For static fields, percent depth dose and profiles were analyzed with criteria in the draft MPPG. The results reveal areas of slight mismatch with the model (MLC leaf penumbra, buildup region.) For TomoTherapy, the IMRT plan 2%/2 mm gamma analysis revealed poorest agreement in the low dose regions. For one static test plan for all 10MV Trilogy photon beams, the plan generation, scan queue creation, data collection, data analysis and report took 2 hours, excluding tank setup. Conclusions: We have demonstrated the implementation feasibility of the TPS MPPG. This exercise generated an open source tool for dose comparisons between scan data and DICOM dose data. An easily reproducible and efficient infrastructure with streamlined data collection was created for repeatable robust testing of the TPS. The tests revealed minor discrepancies in our models and areas for improvement that are being investigated

  20. [Regulation of terpene metabolism]. Annual progress report, March 15, 1991--March 14, 1992

    Energy Technology Data Exchange (ETDEWEB)

    Croteau, R.

    1992-12-31

    This report describes accomplishments over the past year on understanding of terpene synthesis in mint plants and sage. Specifically reported are the fractionation of 4-S-limonene synthetase, the enzyme responsible for the first committed step to monoterpene synthesis, along with isolation of the corresponding RNA and DNA cloning of its gene; the localization of the enzyme within the oil glands, regulation of transcription and translation of the synthetase, the pathway to camphor biosynthesis,a nd studies on the early stages and branch points of the isoprenoid pathway.

  1. The clinical role of the new tumor marker TPS in the follow up of patients with breast cancer. Final report

    International Nuclear Information System (INIS)

    Bakir, M.A.

    1996-08-01

    TPS IRMA is a new assay for quantitative measurement of M3 specific epitope of tissue polypeptide antigen in serum, which occurs especially in conjunction with proliferating tumour cell. The serum TPS concentrations were determined in 34 apparently healthy donors and in 145 patients. Of these patients, 118 had breast cancer and 27 had benign breast diseases. The carcinomas were of different types and included both early (stage I, II) and advanced (stage III, IV) tumours. Clinical stage classification was performed according to TNM system. Blood of neoplastic patients was collected at diagnosis along with serial serum samples during following up in a period of 6-12 months, Serum samples were stored at 30 deg until used. Also, serum levels of two conventional tumour markers CEA (Carcinoembryonic antigen) and CA15-3 were determined for comparison. The distribution of TPS values found in sera from healthy donors ranged between 3 and 60 U/l with a mean of 34±14,57 u/l. We selected a cut-off value for TPS of 63 u/l(X± 2 SD) for healthy controls. Slightly elevated levels of TPS were found in less than 15% of patients with benign breast diseases. High levels of TPS were detected in 85% of patients with breast cancer. The concentration of TPS and the diagnostic sensitivity varied according to stage group. 69% of patients with localised tumours (stage I, II), and 100% of patients with disseminated tumours have shown TPS concentration above the cut-off value. All patients with complete remission have shown TPS values below the cut-off value, in contrast to the progressing group where the values were high. Serial measurements of TPS serum concentration confirmed the fast decrease of TPS where the applied chemotherapy was effective and elevated where the chemotherapy was ineffective. In our study the TPS assay shows a diagnostic sensitivity higher than the CEA and CA15-3 ones. Also, The assay is superior to CEA and CA15-3 ones as TPS reflected earlier the changes of the

  2. Terpene content of wine from the aromatic grape variety ‘Irsai Oliver’ (Vitis vinifera L. depends on maceration time

    Directory of Open Access Journals (Sweden)

    Baron Mojmir

    2017-03-01

    Full Text Available This study deals with the determination of the content of both free and bound terpenes in berries and wine of the aromatic grapevine variety ‘Irsai Oliver’. Grapes were macerated in juice for different time intervals (viz. 0; 5; 12; 24 hours and thereafter processed to wine. The objective was to map the dependence of some selected terpenes on the period of maceration. Using gas chromatography, some nine organic compounds were detected. Attention was paid to contents of linalool (3,7-dimethylokta-1,6-dien-3-ol, 2,6-dimetyl-3,7-octadiene-2,6-diol, hotrienol ([(5E-3,7-dimethylocta-1,5,7-trien-3-yl] acetate, αterpineol (2-(4-Methyl-1-cyclohex-3-enylpropan-2-ol, β-citronellol (3,7-Dimethyloct-6-en-1-ol, nerol ((Z-3,7-dimethyl-2,6-octadien-1-ol, geraniol ((trans-3,7-dimethyl-2,6-oktadien-1-ol and epoxylinalool (2-(5-ethenyl-5-methyloxolan-2-ylpropan-2- ol: epoxylinalool 1 (trans-linalool oxide (furanoid cis-linalool oxide (furanoid and epoxylinalool 2 (trans-linalool oxide (pyranoid cis-linalool oxide (pyranoid. Some basic wine parameters (alcohol, pH, sugars and total acids were estimated as well. The terpene content in wine increased gradually with the period of maceration. The highest and the lowest amounts of terpenes were recorded after 24 hours of maceration and no maceration, respectively. The terpene glycosides content was higher than that of the aglycones. Linalool and 2,6-dimetyl-3,7-octadiene-2,6-diol were the most abundant terpenes.

  3. Potential contribution of fish feed and phytoplankton to the content of volatile terpenes in cultured Pangasius (Pangasianodon hypophthalmus) and Tilapia (Oreochromis niloticus)

    DEFF Research Database (Denmark)

    Podduturi, Raju; Petersen, Mikael Agerlin; Mahmud, Sultan

    2017-01-01

    Geosmin and 2-methylisoborneol are the most recognized off-flavors in freshwater fish, but terpenes may also contribute off-flavor in fish. We identified six monoterpenes, 11 sesquiterpenes, and three terpene-related compounds in pangasius and tilapia from aquaculture farms in Bangladesh. The con...

  4. EKSPERIMEN MODEL PEMBELAJARAN THINK-PAIR-SHARE DENGAN MODUL(TPS-M TERHADAP PRESTASI BELAJAR MATEMATIKA DITINJAU DARI MINAT BELAJAR

    Directory of Open Access Journals (Sweden)

    Jatmiko Jatmiko

    2015-02-01

    Full Text Available Penelitian ini bertujuan untuk mengetahui : (1 manakah diantara Model PembelajaranTPS-M atau Think-Pair-Share” (TPS yang menghasilkan prestasi yang lebih baik, (2 Manakah yang lebih baik, prestasi belajar matematika siswa yang mempunyai minat belajar tinggi, sedang atau rendah, (3 manakah yang menghasilkan prestasi belajar matematika yang lebih baik, di antara Model Pembelajaran TPS-M dan Think-Pair-Share (TPS pada siswa yang mempunyai minat belajar tinggi, sedang dan rendah.Penelitian ini menggunakan metode eksperimental semu (Quasi experimental, dengan desain faktorial 2x3. Populasi penelitian ini adalah semua siswa kelas X SMK kelompok Teknik di Kabupaten Nganjuk Tahun Akademik 2012/2013. Sampel terbagi dalam 2 kelompok, yaitu kelompok eksperimen dan kelompok kontrol. Teknik analisis data yang digunakan dalam penelitian ini, yaitu analisis variansi dua jalan dengan sel tak sama. Berdasarkan hasil analisis diperoleh kesimpulan bahwa: (1 Model pembelajaran TPS-M menghasilkan prestasi belajar matematika yang lebih baik dibandingkan model pembelajaran. (2 Siswa dengan minat belajartinggi, sedang dan rendah memiliki hasil belajar matematika yang sama.(3 Pada masing-masing minat belajar siswa baik tinggi, sedang ataupun rendah prestasi belajar matematika pada model pembelajaran TPS-Mlebih baik dari pada model pembelajaran TPS.

  5. Independent dose calculation of the Tps Iplan in radiotherapy conformed with MLC; Calculo independiente de dosis del TPS Iplan en radioterapia conformada con MLC

    Energy Technology Data Exchange (ETDEWEB)

    Adrada, A.; Tello, Z.; Medina, L.; Garrigo, E.; Venencia, D., E-mail: jorge.alberto.adrada@gmail.com [Instituto Privado de Radioterapia, Obispo Oro 423, X5000BFI Cordoba (Argentina)

    2014-08-15

    The systems utilization of independent dose calculation in three dimensional-Conformal Radiation Therapy (3D-Crt) treatments allows a direct verification of the treatments times. The utilization of these systems allows diminishing the probability of errors occurrence generated by the treatment planning system (Tps), allowing a detailed analysis of the dose to delivering and review of the normalization point (Np) or prescription. The independent dose calculation is realized across the knowledge of dosimetric parameters of the treatment machine and particular characteristics of every individual field. The aim of this work is develops a calculation system of punctual doses for isocentric fields conformed with multi-leaf collimation systems (MLC), where the dose calculation is in conformity with the suggested ones by ICRU Report No. 42, 1987. Calculation software was realized in C ++ under a free platform of programming (Code::Blocks). The system uses files in format Rtp, exported from the Tps to systems of record and verification (Lantis). This file contains detailed information of the dose, Um, position of the MLC sheets and collimators for every field of treatment. The size of equivalent field is obtained from the positions of every sheet; the effective depth of calculation can be introduced from the dosimetric report of the Tps or automatically from the DFS of the field. The 3D coordinates of the isocenter and the Np for the treatment plan must be introduced manually. From this information the system looks the dosimetric parameters and calculates the Um. The calculations were realized in two accelerators a NOVALIS Tx (Varian) with 120 sheets of high definition (hd-MLC) and a PRIMUS Optifocus (Siemens) with 82 sheets. 705 patients were analyzed for a total of 1082, in plans made for both equipment s, the average uncertainty with regard to the calculation of the Tps is-0.43% ± 2.42% in a range between [-7.90 %, 7.50 %]. The major uncertainty was in Np near of the

  6. WE-AB-201-02: TPS Commissioning and QA: A Process Orientation and Application of Control Charts

    Energy Technology Data Exchange (ETDEWEB)

    Sharpe, M. [The Princess Margaret Cancer Centre - UHN (Canada)

    2015-06-15

    Treatment planning systems (TPS) are a cornerstone of modern radiation therapy. Errors in their commissioning or use can have a devastating impact on many patients. To support safe and high quality care, medical physicists must conduct efficient and proper commissioning, good clinical integration, and ongoing quality assurance (QA) of the TPS. AAPM Task Group 53 and related publications have served as seminal benchmarks for TPS commissioning and QA over the past two decades. Over the same time, continuing innovations have made the TPS even more complex and more central to the clinical process. Medical goals are now expressed in terms of the dose and margins around organs and tissues that are delineated from multiple imaging modalities (CT, MR and PET); and even temporally resolved (i.e., 4D) imaging. This information is passed on to optimization algorithms to establish accelerator movements that are programmed directly for IMRT, VMAT and stereotactic treatments. These advances have made commissioning and QA of the TPS much more challenging. This education session reviews up-to-date experience and guidance on this subject; including the recently published AAPM Medical Physics Practice Guideline (MPPG) #5 “Commissioning and QA of Treatment Planning Dose Calculations: Megavoltage Photon and Electron Beams”. Treatment Planning System Commissioning and QA: Challenges and Opportunities (Greg Salomons) This session will provide some key background and review publications describing prominent incidents relating to TPS commissioning and QA. Traditional approaches have been hardware and feature oriented. They aim to establish a functional configuration and establish specifications for regular testing of features (like dose calculation) to assure stable operation and detect failures. With the advent of more complex systems, more patient-specific testing has also been adopted. A number of actual TPS defects will be presented along with heuristics for identifying similar

  7. WE-AB-201-02: TPS Commissioning and QA: A Process Orientation and Application of Control Charts

    International Nuclear Information System (INIS)

    Sharpe, M.

    2015-01-01

    Treatment planning systems (TPS) are a cornerstone of modern radiation therapy. Errors in their commissioning or use can have a devastating impact on many patients. To support safe and high quality care, medical physicists must conduct efficient and proper commissioning, good clinical integration, and ongoing quality assurance (QA) of the TPS. AAPM Task Group 53 and related publications have served as seminal benchmarks for TPS commissioning and QA over the past two decades. Over the same time, continuing innovations have made the TPS even more complex and more central to the clinical process. Medical goals are now expressed in terms of the dose and margins around organs and tissues that are delineated from multiple imaging modalities (CT, MR and PET); and even temporally resolved (i.e., 4D) imaging. This information is passed on to optimization algorithms to establish accelerator movements that are programmed directly for IMRT, VMAT and stereotactic treatments. These advances have made commissioning and QA of the TPS much more challenging. This education session reviews up-to-date experience and guidance on this subject; including the recently published AAPM Medical Physics Practice Guideline (MPPG) #5 “Commissioning and QA of Treatment Planning Dose Calculations: Megavoltage Photon and Electron Beams”. Treatment Planning System Commissioning and QA: Challenges and Opportunities (Greg Salomons) This session will provide some key background and review publications describing prominent incidents relating to TPS commissioning and QA. Traditional approaches have been hardware and feature oriented. They aim to establish a functional configuration and establish specifications for regular testing of features (like dose calculation) to assure stable operation and detect failures. With the advent of more complex systems, more patient-specific testing has also been adopted. A number of actual TPS defects will be presented along with heuristics for identifying similar

  8. PERBEDAAN PRESTASI BELAJAR PEMROGRAMAN DASAR PENERAPAN PJBL DIBANDINGKAN TPS PADA SISWA KELAS X TKJ DI SMK

    Directory of Open Access Journals (Sweden)

    Lia Nur Enis Ratna Wijayanti

    2016-02-01

    Full Text Available Tujuan penelitian untuk mengungkap perbedaan prestasi belajar Pemro-graman Dasar materi Algoritma Percabangan siswa yang diajar menggunakan model pembelajaran Project Based Learning (PjBL dan Think pair Share (TPS. Rancang-an penelitian menggunakan pretest-posttest control group design. Subjek penelitian adalah siswa Kelas X Paket Keahlian TKJ di SMK Negeri 2 Singosari Malang seba-nyak 67 siswa. Teknik sampling yang digunakan adalah sampling jenuh, karena selu-ruh anggota populasi digunakan sebagai sampel. Instrumen penelitian menggunakan instrumen perlakuan dan pengukuran. Teknik analisis data: (1 uji prasyarat (normali-tas, homogenitas, dan uji kesamaan dua rerata kemampuan awal; dan (2 uji hipotesis (uji-t. Hasil penelitian menunjukkan ada perbedaan prestasi belajar yang signifikan antara kelas PjBL dan TPS.

  9. Independent dose calculation in IMRT for the Tps Iplan using the Clarkson modified integral

    International Nuclear Information System (INIS)

    Adrada, A.; Tello, Z.; Garrigo, E.; Venencia, D.

    2014-08-01

    Intensity-Modulated Radiation Therapy (IMRT) treatments require a quality assurance (Q A) specific patient before delivery. These controls include the experimental verification in dose phantom of the total plan as well as dose distributions. The use of independent dose calculation (IDC) is used in 3D-Crt treatments; however its application in IMRT requires the implementation of an algorithm that allows considering a non-uniform intensity beam. The purpose of this work was to develop IDC software in IMRT with MLC using the algorithm proposed by Kung (Kung et al. 2000). The software was done using Matlab programming. The Clarkson modified integral was implemented on each flowing, applying concentric rings for the dose determination. From the integral of each field was calculated the dose anywhere. One time finished a planning; all data are exported to a phantom where a Q A plan is generated. On this is calculated the half dose in a representative volume of the ionization chamber and the dose at the center of it. Until now 230 IMRT planning were analyzed carried out ??in the treatment planning system (Tps) Iplan. For each one of them Q A plan was generated, were calculated and compared calculated dose with the Tps, IDC system and measurement with ionization chamber. The average difference between measured and calculated dose with the IDC system was 0.4% ± 2.2% [-6.8%, 6.4%]. The difference between the measured and the calculated doses by the pencil-beam algorithm (Pb) of Tps was 2.6% ± 1.41% [-2.0%, 5.6%] and with the Monte Carlo algorithm was 0.4% ± 1.5% [-4.9%, 3.7%]. The differences of the carried out software are comparable to the obtained with the ionization chamber and Tps in Monte Carlo mode. (author)

  10. Evaluation of six TPS algorithms in computing entrance and exit doses

    Science.gov (United States)

    Metwaly, Mohamed; Glegg, Martin; Baggarley, Shaun P.; Elliott, Alex

    2014-01-01

    Entrance and exit doses are commonly measured in in vivo dosimetry for comparison with expected values, usually generated by the treatment planning system (TPS), to verify accuracy of treatment delivery. This report aims to evaluate the accuracy of six TPS algorithms in computing entrance and exit doses for a 6 MV beam. The algorithms tested were: pencil beam convolution (Eclipse PBC), analytical anisotropic algorithm (Eclipse AAA), AcurosXB (Eclipse AXB), FFT convolution (XiO Convolution), multigrid superposition (XiO Superposition), and Monte Carlo photon (Monaco MC). Measurements with ionization chamber (IC) and diode detector in water phantoms were used as a reference. Comparisons were done in terms of central axis point dose, 1D relative profiles, and 2D absolute gamma analysis. Entrance doses computed by all TPS algorithms agreed to within 2% of the measured values. Exit doses computed by XiO Convolution, XiO Superposition, Eclipse AXB, and Monaco MC agreed with the IC measured doses to within 2%‐3%. Meanwhile, Eclipse PBC and Eclipse AAA computed exit doses were higher than the IC measured doses by up to 5.3% and 4.8%, respectively. Both algorithms assume that full backscatter exists even at the exit level, leading to an overestimation of exit doses. Despite good agreements at the central axis for Eclipse AXB and Monaco MC, 1D relative comparisons showed profiles mismatched at depths beyond 11.5 cm. Overall, the 2D absolute gamma (3%/3 mm) pass rates were better for Monaco MC, while Eclipse AXB failed mostly at the outer 20% of the field area. The findings of this study serve as a useful baseline for the implementation of entrance and exit in vivo dosimetry in clinical departments utilizing any of these six common TPS algorithms for reference comparison. PACS numbers: 87.55.‐x, 87.55.D‐, 87.55.N‐, 87.53.Bn PMID:24892349

  11. Cabling design of booster and storage ring construction progress of TPS

    International Nuclear Information System (INIS)

    Wong, Y.-S.; Liu, K.-B.; Liu, C.-Y.; Wang, B.-S.

    2017-01-01

    The 2012 Taiwan Photon Source (TPS) cable construction project started after 10 months to complete the cable laying and installation of power supply. The circumference of the booster ring (BR) is 496.8 m, whereas that of the storage ring (SR) is 518.4 m. Beam current is set to 500 mA at 3.3 GeV. The paper on grounding systems discusses the design of the ground wire (< 0.2 Ω) with low impedance, power supply of the accelerator and cabling tray. The flow and size of the ground current are carefully evaluated to avoid grounded current from flowing everywhere, which causes interference problems. In the design of the TPS, special shielding will be established to isolate the effects of electromagnetic interference on the magnet and ground current. Booster ring dipoles are connected by a series of 54-magnet bending dipole; the cable size of its stranded wire measures 250 mm"2, with a total length of 5000 m. Booster ring and storage ring quadrupoles have 150 magnets; the cable size of their stranded wire is 250 mm"2, with a total length of 17000 m. Storage ring dipole consists of 48 magnets; the cable size of its stranded wire is 325 mm"2, with a total length of 6000 m. This study discusses the power supply cabling design of the storage ring and booster ring construction progress of TPS. The sections of this paper are divided into discussions of the construction of the control and instrument area, cabling layout of booster ring and storage ring, as well as the installation and commission machine. This study also discusses the use of a high-impedance meter to determine the effect of cabling insulation and TPS power supply machine on energy transfer to ensure the use of safe and correct magnet.

  12. In Depth Analysis of AVCOAT TPS Response to a Reentry Flow

    International Nuclear Information System (INIS)

    Titov, E. V.; Kumar, Rakesh; Levin, D. A.

    2011-01-01

    Modeling of the high altitude portion of reentry vehicle trajectories with DSMC or statistical BGK solvers requires accurate evaluation of the boundary conditions at the ablating TPS surface. Presented in this article is a model which takes into account the complex ablation physics including the production of pyrolysis gases, and chemistry at the TPS surface. Since the ablation process is time dependent the modeling of the material response to the high energy reentry flow starts with the solution of the rarefied flow over the vehicle and then loosely couples with the material response. The objective of the present work is to carry out conjugate thermal analysis by weakly coupling a flow solver to a material thermal response model. The latter model solves the one dimensional heat conduction equation accounting for the pyrolysis process that takes place in the reaction zone of an ablative thermal protection system (TPS) material. An estimate of the temperature range within which the pyrolysis reaction (decomposition and volatilization) takes place is obtained from Ref. [1]. The pyrolysis reaction results in the formation of char and the release of gases through the porous charred material. These gases remove additional amount of heat as they pass through the material, thus cooling the material (the process known as transpiration cooling). In the present work, we incorporate the transpiration cooling model in the material thermal response code in addition to the pyrolysis model. The flow in the boundary layer and in the vicinity of the TPS material is in the transitional flow regime. Therefore, we use a previously validated statistical BGK method to model the flow physics in the vicinity of the micro-cracks, since the BGK method allows simulations of flow at pressures higher than can be computed using DSMC.

  13. A Statistics-Based Material Property Analysis to Support TPS Characterization

    Science.gov (United States)

    Copeland, Sean R.; Cozmuta, Ioana; Alonso, Juan J.

    2012-01-01

    Accurate characterization of entry capsule heat shield material properties is a critical component in modeling and simulating Thermal Protection System (TPS) response in a prescribed aerothermal environment. The thermal decomposition of the TPS material during the pyrolysis and charring processes is poorly characterized and typically results in large uncertainties in material properties as inputs for ablation models. These material property uncertainties contribute to large design margins on flight systems and cloud re- construction efforts for data collected during flight and ground testing, making revision to existing models for entry systems more challenging. The analysis presented in this work quantifies how material property uncertainties propagate through an ablation model and guides an experimental test regimen aimed at reducing these uncertainties and characterizing the dependencies between properties in the virgin and charred states for a Phenolic Impregnated Carbon Ablator (PICA) based TPS. A sensitivity analysis identifies how the high-fidelity model behaves in the expected flight environment, while a Monte Carlo based uncertainty propagation strategy is used to quantify the expected spread in the in-depth temperature response of the TPS. An examination of how perturbations to the input probability density functions affect output temperature statistics is accomplished using a Kriging response surface of the high-fidelity model. Simulations are based on capsule configuration and aerothermal environments expected during the Mars Science Laboratory (MSL) entry sequence. We identify and rank primary sources of uncertainty from material properties in a flight-relevant environment, show the dependence on spatial orientation and in-depth location on those uncertainty contributors, and quantify how sensitive the expected results are.

  14. Screening Study of Leaf Terpene Concentration of 75 Borneo Rainforest Plant Species: Relationships with Leaf Elemental Concentrations and Morphology

    Directory of Open Access Journals (Sweden)

    Jordi Sardans

    2015-01-01

    Full Text Available Terpenes confer advantage in plant protection against abiotic stresses such as heat and drought and biotic stresses such as herbivore and pathogen attack. We conducted a screening of leaf mono- and sesquiterpene concentrations in 75 common woody plant species in the rainforest of Danum Valley (Borneo. Terpene compounds were found in 73 out of the 75 analysed species. Similar or lower proportions have been reported in other parts of the world. To our knowledge, this study reports for the first time the foliar concentration of mono- and/or sesquiterpene for 71 species and 39 genera not previously analyzed. Altogether 80 terpene compounds were determined across the species, and out of these only linalool oxide and (E- g -bisabolene had phylogenetic signal. A significant negative relationship between leaf monoterpene concentration and leaf length was observed, but leaf mono- and sesquitepene concentration were not related to any other leaf morphological trait nor to leaf elemental composition. Functions such as temperature protection, radiation protection or signaling and communication could underlie the high frequency of terpene-containing species of this tropical ecosystem which has multiple and very diverse interactions among multiple species.

  15. Surface chemistry of a pine-oil cleaner and other terpene mixtures with ozone on vinyl flooring tiles.

    Science.gov (United States)

    Ham, Jason E; Wells, J Raymond

    2011-04-01

    Indoor environments are dynamic reactors where consumer products (such as cleaning agents, deodorants, and air fresheners) emit volatile organic compounds (VOCs) that can subsequently interact with indoor oxidants such as ozone (O(3)), hydroxyl radicals, and nitrate radicals. Typically, consumer products consist of mixtures of VOCs and semi-VOCs which can react in the gas-phase or on surfaces with these oxidants to generate a variety of oxygenated products. In this study, the reaction of a pine-oil cleaner (POC) with O(3) (100ppb) on a urethane-coated vinyl flooring tile was investigated at 5% and 50% relative humidity. These results were compared to previous α-terpineol+O(3) reactions on glass and vinyl surfaces. Additionally, other terpene and terpene alcohol mixtures were formulated to understand the emission profiles as seen in the POC data. Results showed that the α-terpineol+O(3) reaction products were the prominent species that were also observed in the POC/O(3) surface experiments. Furthermore, α-terpineol+O(3) reactions generate the largest fraction of oxygenated products even in equal mixtures of other terpene alcohols. This finding suggests that the judicial choice of terpene alcohols for inclusion in product formulations may be useful in reducing oxidation product emissions. Published by Elsevier Ltd.

  16. Nonlinear Aeroelastic Analysis of the HIAD TPS Coupon in the NASA 8' High Temperature Tunnel: Theory and Experiment

    Science.gov (United States)

    Goldman, Benjamin D.; Scott, Robert C,; Dowell, Earl H.

    2014-01-01

    The purpose of this work is to develop a set of theoretical and experimental techniques to characterize the aeroelasticity of the thermal protection system (TPS) on the NASA Hypersonic Inflatable Aerodynamic Decelerator (HIAD). A square TPS coupon experiences trailing edge oscillatory behavior during experimental testing in the 8' High Temperature Tunnel (HTT), which may indicate the presence of aeroelastic flutter. Several theoretical aeroelastic models have been developed, each corresponding to a different experimental test configuration. Von Karman large deflection theory is used for the plate-like components of the TPS, along with piston theory for the aerodynamics. The constraints between the individual TPS layers and the presence of a unidirectional foundation at the back of the coupon are included by developing the necessary energy expressions and using the Rayleigh Ritz method to derive the nonlinear equations of motion. Free vibrations and limit cycle oscillations are computed and the frequencies and amplitudes are compared with accelerometer and photogrammetry data from the experiments.

  17. Possibility for implementation of e-beam technology in TPS Sviloza

    International Nuclear Information System (INIS)

    Dutskinov, N.; Pelovski, Y.; Nikolov, K.; Stamatov, L.

    2011-01-01

    The electron beam flue gas treatment process is one of the most promising technologies in the modern environmental protection. The technology allows the simultaneous removal of acidic pollutants such as SO 2 and NO x with high efficiency and decomposition of VOC (volatile organic compounds) without generating any wastes. These pollutants are named “acid gases” that cause acid rain and damage forests, agriculture fields and flora as well as public health. After successful operation of Electron Beam Flue Gas Treatment Pilot Plant at Maritsa East 2 TPS, the Bulgarian Ministry of Economy and Energy has taken decision for implementation EB technology in the industrial scale at “Sviloza” TPS in Svishtov, Bulgaria. The Industrial Electron Beam Flue Gas Treatment Plant (IEBFGTP) covers 100% of the flue gases generated from all units of TPS “Sviloza” JSC, Svishtov. This Thermal Power Station generates flue gases from all boilers – 600 000 Nm3/h, with emission of SO 2 → 2800 – 4800 mg/Nm 3 , NOx → 1200 – 1600 mg/Nm 3 and dust → 200 – 1400 mg/Nm3. The major objective of the electron beam project is to reduce harmful emissions of SO x , NO x and VOC by 85%. The byproduct generated by the electron beam plant is ammonium sulfate and ammonium nitrate and it can be used as fertilizer in the Bulgarian agriculture sector and abroad. (author)

  18. Laser Shearography Inspection of TPS (Thermal Protection System) Cork on RSRM (Reusable Solid Rocket Motors)

    Science.gov (United States)

    Lingbloom, Mike; Plaia, Jim; Newman, John

    2006-01-01

    Laser Shearography is a viable inspection method for detection of de-bonds and voids within the external TPS (thermal protection system) on to the Space Shuttle RSRM (reusable solid rocket motors). Cork samples with thicknesses up to 1 inch were tested at the LTI (Laser Technology Incorporated) laboratory using vacuum-applied stress in a vacuum chamber. The testing proved that the technology could detect cork to steel un-bonds using vacuum stress techniques in the laboratory environment. The next logical step was to inspect the TPS on a RSRM. Although detailed post flight inspection has confirmed that ATK Thiokol's cork bonding technique provides a reliable cork to case bond, due to the Space Shuttle Columbia incident there is a great interest in verifying bond-lines on the external TPS. This interest provided and opportunity to inspect a RSRM motor with Laser Shearography. This paper will describe the laboratory testing and RSRM testing that has been performed to date. Descriptions of the test equipment setup and techniques for data collection and detailed results will be given. The data from the test show that Laser Shearography is an effective technology and readily adaptable to inspect a RSRM.

  19. Possibility for implementation of e-beam technology in TPS Sviloza

    Energy Technology Data Exchange (ETDEWEB)

    Dutskinov, N. [Ministry of Energy, NEK, Sofia (Bulgaria); Pelovski, Y. [University of Chemical Technology and Metallurgy, Sofia (Bulgaria); Nikolov, K.; Stamatov, L. [Sviloza Power Station, Svishtov (Bulgaria)

    2011-07-01

    The electron beam flue gas treatment process is one of the most promising technologies in the modern environmental protection. The technology allows the simultaneous removal of acidic pollutants such as SO{sub 2} and NO{sub x} with high efficiency and decomposition of VOC (volatile organic compounds) without generating any wastes. These pollutants are named “acid gases” that cause acid rain and damage forests, agriculture fields and flora as well as public health. After successful operation of Electron Beam Flue Gas Treatment Pilot Plant at Maritsa East 2 TPS, the Bulgarian Ministry of Economy and Energy has taken decision for implementation EB technology in the industrial scale at “Sviloza” TPS in Svishtov, Bulgaria. The Industrial Electron Beam Flue Gas Treatment Plant (IEBFGTP) covers 100% of the flue gases generated from all units of TPS “Sviloza” JSC, Svishtov. This Thermal Power Station generates flue gases from all boilers – 600 000 Nm3/h, with emission of SO{sub 2} → 2800 – 4800 mg/Nm{sup 3}, NOx → 1200 – 1600 mg/Nm{sup 3} and dust → 200 – 1400 mg/Nm3. The major objective of the electron beam project is to reduce harmful emissions of SO{sub x}, NO{sub x} and VOC by 85%. The byproduct generated by the electron beam plant is ammonium sulfate and ammonium nitrate and it can be used as fertilizer in the Bulgarian agriculture sector and abroad. (author)

  20. Producing biofuels using polyketide synthases

    Science.gov (United States)

    Katz, Leonard; Fortman, Jeffrey L; Keasling, Jay D

    2013-04-16

    The present invention provides for a non-naturally occurring polyketide synthase (PKS) capable of synthesizing a carboxylic acid or a lactone, and a composition such that a carboxylic acid or lactone is included. The carboxylic acid or lactone, or derivative thereof, is useful as a biofuel. The present invention also provides for a recombinant nucleic acid or vector that encodes such a PKS, and host cells which also have such a recombinant nucleic acid or vector. The present invention also provides for a method of producing such carboxylic acids or lactones using such a PKS.

  1. Assessing terpene content variability of whitebark pine in order to estimate representative sample size

    Directory of Open Access Journals (Sweden)

    Stefanović Milena

    2013-01-01

    Full Text Available In studies of population variability, particular attention has to be paid to the selection of a representative sample. The aim of this study was to assess the size of the new representative sample on the basis of the variability of chemical content of the initial sample on the example of a whitebark pine population. Statistical analysis included the content of 19 characteristics (terpene hydrocarbons and their derivates of the initial sample of 10 elements (trees. It was determined that the new sample should contain 20 trees so that the mean value calculated from it represents a basic set with a probability higher than 95 %. Determination of the lower limit of the representative sample size that guarantees a satisfactory reliability of generalization proved to be very important in order to achieve cost efficiency of the research. [Projekat Ministarstva nauke Republike Srbije, br. OI-173011, br. TR-37002 i br. III-43007

  2. Impact of biogenic terpene emissions from Brassica napus on tropospheric ozone over Saxony (Germany): numerical investigation.

    Science.gov (United States)

    Renner, Eberhard; Münzenberg, Annette

    2003-01-01

    The role of biogenic emissions in tropospheric ozone production is currently under discussion and major aspects are not well understood yet. This study aims towards the estimation of the influence of biogenic emissions on tropospheric ozone concentrations over Saxony in general and of biogenic emissions from brassica napus in special. MODELLING TOOLS: The studies are performed by utilizing a coupled numerical modelling system consisting of the meteorological model METRAS and the chemistry transport model MUSCAT. For the chemical part, the Euro-RADM algorithm is used. EMISSIONS: Anthropogenic and biogenic emissions are taken into account. The anthropogenic emissions are introduced by an emission inventory. Biogenic emissions, VOC and NO, are calculated within the chemical transport model MUSCAT at each time step and in each grid cell depending on land use type and on the temperature. The emissions of hydrocarbons from forest areas as well as biogenic NO especially from agricultural grounds are considered. Also terpene emissions from brassica napus fields are estimated. SIMULATION SETUP AND METEOROLOGICAL CONDITIONS: The simulations were performed over an area with an extension of 160 x 140 km2 which covers the main parts of Saxony and neighboring areas of Brandenburg, Sachsen-Anhalt and Thuringia. Summer smog with high ozone concentrations can be expected during high pressure conditions on hot summer days. Typical meteorological conditions for such cases were introduced in an conceptual way. It is estimated that biogenic emissions change tropospheric ozone concentrations in a noticeable way (up to 15% to 20%) and, therefore, should not be neglected in studies about tropospheric ozone. Emissions from brassica napus do have a moderate potential to enhance tropospheric ozone concentrations, but emissions are still under consideration and, therefore, results vary to a high degree. Summing up, the effect of brassica napus terpene emissions on ozone concentrations is

  3. Application of terpene-induced cell for enhancing biodegradation of TCE contaminated soil

    Directory of Open Access Journals (Sweden)

    Ekawan Luepromchai

    2004-02-01

    Full Text Available Trichloroethylene (TCE, a chlorinated solvent, is a major water pollutant originating from spillage and inappropriate disposal of dry cleaning agents, degreasing solvents, and paint strippers. Due to its widespread contamination and potential health threat, remediation technology to clean-up TCE is necessary. Aerobic biodegradation of TCE is reported to occur via cometabolism, by which TCE degrading bacteria utilize other compounds such as toluene, phenol, and methane as growth substrate and enzyme inducer. Although toluene is reported to be the most effective inducer, it is regulated as a hazardous material and should not be applied to the environment. The objectives of this study were to identify an alternative enzyme inducer as well as to apply the induced bacteria for degradation of TCE in contaminated soil. We investigated the effect of terpenes, the main components in volatile essential oils of plants, on induction of TCE degradation in Rhodococcus gordoniae P3, a local Gram (+ bacterium. Selected terpenes including cumene, limonene, carvone and pinene at various concentrations were used in the study. Results from liquid culture showed that 25 mg l-1 cumeneinduced R. gordoniae P3 cells resulted in 75% degradation of 10 ppm TCE within 24 hrs. Soil microcosms were later employed to investigate the ability of cumene to enhance TCE biodegradation in the environment. There were two bioremediation treatments studied, including bioaugmentation, the inoculation of cumeneinduced R. gordoniae P3, and biostimulation, the addition of cumene to induce soil indigenous microorganisms to degrade TCE. Bioaugmentation and biostimulation were shown to accelerate TCE reduction significantly more than control treatment at the beginning of study. The results suggest that cumene-induced R. gordoniae P3 and cumene can achieve rapid TCE biodegradation.

  4. Heterooligomeric phosphoribosyl diphosphate synthase of Saccharomyces cerevisiae

    DEFF Research Database (Denmark)

    Hove-Jensen, Bjarne

    2004-01-01

    The yeast Saccharomyces cerevisiae contains five phosphoribosyl diphosphate (PRPP) synthase-homologous genes (PRS1-5), which specify PRPP synthase subunits 1-5. Expression of the five S. cerevisiae PRS genes individually in an Escherichia coli PRPP-less strain (Deltaprs) showed that a single PRS...

  5. Threonine phosphorylation of rat liver glycogen synthase

    International Nuclear Information System (INIS)

    Arino, J.; Arro, M.; Guinovart, J.J.

    1985-01-01

    32 P-labeled glycogen synthase specifically immunoprecipitated from 32 P-phosphate incubated rat hepatocytes contains, in addition to [ 32 P] phosphoserine, significant levels of [ 32 P] phosphothreonine. When the 32 P-immunoprecipitate was cleaved with CNBr, the [ 32 P] phosphothreonine was recovered in the large CNBr fragment (CB-2, Mapp 28 Kd). Homogeneous rat liver glycogen synthase was phosphorylated by all the protein kinases able to phosphorylate CB-2 in vitro. After analysis of the immunoprecipitated enzyme for phosphoaminoacids, it was observed that only casein kinase II was able to phosphorylate on threonine and 32 P-phosphate was only found in CB-2. These results demonstrate that rat liver glycogen synthase is phosphorylated at threonine site(s) contained in CB-2 and strongly indicate that casein kinase II may play a role in the ''in vivo'' phosphorylation of liver glycogen synthase. This is the first protein kinase reported to phosphorylate threonine residues in liver glycogen synthase

  6. PERBANDINGAN METODE PEMBELAJARAN NUMBERED HEAD TOGETHER (NHT DENGAN THINK PAIR SHARE (TPS PADA HASIL BELAJAR KEWIRAUSAHAAN

    Directory of Open Access Journals (Sweden)

    M. Maulani Subhi

    2016-11-01

    Full Text Available Pendidikan berintikan interaksi pendidik dengan peserta didik dalam upaya membantu peserta didik menguasai tujuan-tujuan pendidikan. Pendidikan akan berhasil, jika proses pembelajaran berlangsung dengan baik. Proses pembelajaran akan berlangsung baik apabila metode pembelajaran yang diterapkan sesuai dengan keadaan siswa. Selama ini proses pembelajaran masih dilakukan secara konvensional. Sebagai akibatnya, terlihat pada hasil observasi awal hasil belajar siswa di SMK Islam Roudlotus Saidiyyah masih banyak siswa yang belum tuntas nilainya. Untuk menghindari hal tersebut, salah satu upayanya dengan menerapkan Metode Pembelajaran Numbered Head Together (NHT dan Think Pair Share (TPS. Rumusan masalah dalam penelitian adalah Adakah perbedaan hasil belajar antara metode pembelajaranNumbered Head Together (NHT dengan Think Pair Share (TPS terhadap hasil belajar mata pelajaran kewirausahaan kelas X SMK Islam Roudlotus Saidiyyah Semarang.Metode penelitian yang digunakan adalah quasi eksperimen. Populasi dalam penelitian ini adalah seluruh kelas X SMK Islam Roudlotus Saidiyyah Semarang. Teknik pengambilan sampel menggunakan teknik sampling kuota, sehingga mengambil sampel 32 siswa, kelas X TKJ sebagai kelas eksperimen pertema dan kelas X PS sebagai kelas eksperimen kedua. Teknik pengumpulan data menggunakan dokumentasi, tes dan angket dan analisis data menggunakan uji t.Berdasarkan hasil perhitungan didapatkan nilai Sig. (2-tailed untuk motivasi siswa 0,009, nilai Sig. (2-tailed untuk respon siswa 0,035, nilai Sig.(2-tailed untuk hasil belajar siswa 0,03. Berdasarkan nilai Sig. (2-tailed tersebut dapat disimpulkan bahwa terdapat perbedaan hasil belajar siswa kelas NHT dengan kelas TPS. Rata-rata tingkat motivasi siswa 73,8%, tingkat respon siswa72,0%, dan hasil belajar siswa 71,0% pada kelas NHT lebih tinggi dari pada kelas TPS dengan rata-rata tingkat motivasi siswa 57,5%, tingkat respon siswa 61,5%, dan hasil belajar siswa 67,8%.Dari hasil penelitian

  7. Identification of the trehalose-6-phosphate synthase gene family in ...

    Indian Academy of Sciences (India)

    2015-03-04

    Mar 4, 2015 ... stress, however, our study mainly analysed the TPS gene family under freezing conditions in winter wheat .... size the first-strand cDNA using the Fermentas RevertAid ..... In the stem of Dongnongdongmai 1, TaTPS1, 2, 3, 4, 8,.

  8. ESTUDIO FISICOQUÍMICO DE MEZCLAS DE ALMIDÓN TERMOPLÁSTICO (TPS Y POLICAPROLACTONA (PCL

    Directory of Open Access Journals (Sweden)

    JOSE MINA H.

    Full Text Available Se estudiaron las propiedades físico-químicas de un almidón termoplástico (TPS de yuca, plastificado con glicerol, y el efecto de la incorporación de policaprolactona (PCL en el desarrollo de tres mezclas binarias TPS-PCL a proporciones 60-40, 50-50 y 40-60. El estudio inició con la preparación por extrusión del TPS y su posterior mezcla en caliente con la policaprolactona. El análisis de los resultados se enfocó principalmente a la identificación y seguimiento de la retrogradación, que se dio en el almidón termoplástico para diferentes tiempos de acondicionamiento (a 54% H.R. y 25ºC. Con el proceso de plastificación del almidón se generó una masa predominantemente amorfa, evidenciada por los resultados obtenidos con SEM, DRX y FTIR. Con la incorporación de la PCL se obtuvieron mezclas inmiscibles, en las cuales el TPS se constituyó en la fase dispersa y que conservaron una tendencia en la variación de las propiedades con el tiempo de acondicionamiento, similar a la observada en el TPS solo; indicando este hecho que las interacciones físico-químicas generadas entre el TPS y la policaprolactona no fueron lo suficientemente fuertes, como para incidir en la variación estructural del TPS, la cual se dio independiente del contenido de PCL.

  9. Glycogen Synthase Kinase-3β

    DEFF Research Database (Denmark)

    Munkholm, Klaus; Lenskjold, Toke; Jacoby, Anne Sophie

    2016-01-01

    cells were quantitated using enzyme immunometric assays. The activity of GSK-3β (serine-9-phosphorylated GSK-3β/total GSK-3β) was lower at baseline compared with follow-up. No significant mean change over time was observed in levels of total GSK-3β and serine-9-phosphorylated GSK-3β. Exploratory......Evidence indicates a role for glycogen synthase kinase-3β (GSK-3β) in the pathophysiology of mood disorders and in cognitive disturbances; however, the natural variation in GSK-3β activity over time is unknown. We aimed to investigate GSK-3β activity over time and its possible correlation...... with emotional lability, subjective mood fluctuations and cognitive function in healthy individuals. Thirty-seven healthy subjects were evaluated with neuropsychological tests and blood samples at baseline and 12-week follow-up. Total GSK-3β and serine-9-phosphorylated GSK-3β in peripheral blood mononuclear...

  10. BrachyTPS -Interactive point kernel code package for brachytherapy treatment planning of gynaecological cancers

    International Nuclear Information System (INIS)

    Thilagam, L.; Subbaiah, K.V.

    2008-01-01

    Brachytherapy treatment planning systems (TPS) are always recommended to account for the effect of tissue, applicator and shielding material heterogeneities exist in Intracavitary brachytherapy (ICBT) applicators. Most of the commercially available brachytherapy TPS softwares estimate the absorbed dose at a point, only taking care of the contributions of individual sources and the source distribution, neglecting the dose perturbations arising from the applicator design and construction. So the doses estimated by them are not much accurate under realistic clinical conditions. In this regard, interactive point kernel rode (BrachyTPS) has been developed to perform independent dose calculations by taking into account the effect of these heterogeneities, using two regions build up factors, proposed by Kalos. As primary input data, the code takes patients' planning data including the source specifications, dwell positions, dwell times and it computes the doses at reference points by dose point kernel formalisms, with multi-layer shield build-up factors accounting for the contributions from scattered radiation. In addition to performing dose distribution calculations, this code package is capable of displaying an isodose distribution curve into the patient anatomy images. The primary aim of this study is to validate the developed point kernel code integrated with treatment planning systems against the other tools which are available in the market. In the present work, three brachytherapy applicators commonly used in the treatment of uterine cervical carcinoma, Board of Radiation Isotope and Technology (BRIT) made low dose rate (LDR) applicator, Fletcher Green type LDR applicator and Fletcher Williamson high dose rate (HDR) applicator were studied to test the accuracy of the software

  11. A method to incorporate interstitial components into the TPS gynecologic rigid applicator library

    Directory of Open Access Journals (Sweden)

    Antonio Otal

    2017-01-01

    Full Text Available Purpose: T2 magnetic resonance imaging (MRI is recommended as the imaging modality for image-guided brachytherapy. In locally advanced cervical carcinoma, combined endocavitary and interstitial applicators are appropriate (Vienna or Utrecht. To cover extensive disease, Template Benidorm (TB was developed. Treatment planning system applicator libraries are currently unavailable for the Utrecht applicator or the TB. The purpose of this work is to develop an applicator library for both applicators. Material and methods: The library developed in this work has been used in the Oncentra Brachytherapy TPS, version 4.3.0, which has a brachytherapy module that includes a library of rigid applicators. To add the needles of the Utrecht applicator and to model the TB, we used FreeCAD and MeshLab. The reconstruction process was based on the points that the rigid section and the interstitial part have in common. This, together with the free length, allowed us to ascertain the position of the tip. Results: In case of the Utrecht applicator, one of the sources of uncertainty in the reconstruction was determining the distance of the tip of needle from the ovoid. In case of the TB, the large number of needles involved made their identification time consuming. The developed library resolved both issues. Conclusions : The developed library for the Utrecht and TB is feasible and efficient improving accuracy. It allows all the required treatment planning to proceed using just a T2 MRI sequence. The additional use of specific free available software applications makes it possible to add this information to the already existing library of the Oncentra Brachytherapy TPS. Specific details not included on this manuscript will be available under request. This library is also currently being implemented also into the Sagiplan v 2.0 TPS.

  12. Crystal structure of riboflavin synthase

    Energy Technology Data Exchange (ETDEWEB)

    Liao, D.-I.; Wawrzak, Z.; Calabrese, J.C.; Viitanen, P.V.; Jordan, D.B. (DuPont); (NWU)

    2010-03-05

    Riboflavin synthase catalyzes the dismutation of two molecules of 6,7-dimethyl-8-(1'-D-ribityl)-lumazine to yield riboflavin and 4-ribitylamino-5-amino-2,6-dihydroxypyrimidine. The homotrimer of 23 kDa subunits has no cofactor requirements for catalysis. The enzyme is nonexistent in humans and is an attractive target for antimicrobial agents of organisms whose pathogenicity depends on their ability to biosynthesize riboflavin. The first three-dimensional structure of the enzyme was determined at 2.0 {angstrom} resolution using the multiwavelength anomalous diffraction (MAD) method on the Escherichia coli protein containing selenomethionine residues. The homotrimer consists of an asymmetric assembly of monomers, each of which comprises two similar {beta} barrels and a C-terminal {alpha} helix. The similar {beta} barrels within the monomer confirm a prediction of pseudo two-fold symmetry that is inferred from the sequence similarity between the two halves of the protein. The {beta} barrels closely resemble folds found in phthalate dioxygenase reductase and other flavoproteins. The three active sites of the trimer are proposed to lie between pairs of monomers in which residues conserved among species reside, including two Asp-His-Ser triads and dyads of Cys-Ser and His-Thr. The proposed active sites are located where FMN (an analog of riboflavin) is modeled from an overlay of the {beta} barrels of phthalate dioxygenase reductase and riboflavin synthase. In the trimer, one active site is formed, and the other two active sites are wide open and exposed to solvent. The nature of the trimer configuration suggests that only one active site can be formed and be catalytically competent at a time.

  13. Independent dose calculation of the Tps Iplan in radiotherapy conformed with MLC

    International Nuclear Information System (INIS)

    Adrada, A.; Tello, Z.; Medina, L.; Garrigo, E.; Venencia, D.

    2014-08-01

    The systems utilization of independent dose calculation in three dimensional-Conformal Radiation Therapy (3D-Crt) treatments allows a direct verification of the treatments times. The utilization of these systems allows diminishing the probability of errors occurrence generated by the treatment planning system (Tps), allowing a detailed analysis of the dose to delivering and review of the normalization point (Np) or prescription. The independent dose calculation is realized across the knowledge of dosimetric parameters of the treatment machine and particular characteristics of every individual field. The aim of this work is develops a calculation system of punctual doses for isocentric fields conformed with multi-leaf collimation systems (MLC), where the dose calculation is in conformity with the suggested ones by ICRU Report No. 42, 1987. Calculation software was realized in C ++ under a free platform of programming (Code::Blocks). The system uses files in format Rtp, exported from the Tps to systems of record and verification (Lantis). This file contains detailed information of the dose, Um, position of the MLC sheets and collimators for every field of treatment. The size of equivalent field is obtained from the positions of every sheet; the effective depth of calculation can be introduced from the dosimetric report of the Tps or automatically from the DFS of the field. The 3D coordinates of the isocenter and the Np for the treatment plan must be introduced manually. From this information the system looks the dosimetric parameters and calculates the Um. The calculations were realized in two accelerators a NOVALIS Tx (Varian) with 120 sheets of high definition (hd-MLC) and a PRIMUS Optifocus (Siemens) with 82 sheets. 705 patients were analyzed for a total of 1082, in plans made for both equipment s, the average uncertainty with regard to the calculation of the Tps is-0.43% ± 2.42% in a range between [-7.90 %, 7.50 %]. The major uncertainty was in Np near of the

  14. De novo production of six key grape aroma monoterpenes by a geraniol synthase-engineered S. cerevisiae wine strain.

    Science.gov (United States)

    Pardo, Ester; Rico, Juan; Gil, José Vicente; Orejas, Margarita

    2015-09-16

    Monoterpenes are important contributors to grape and wine aroma. Moreover, certain monoterpenes have been shown to display health benefits with antimicrobial, anti-inflammatory, anticancer or hypotensive properties amongst others. The aim of this study was to construct self-aromatizing wine yeasts to overproduce de novo these plant metabolites in wines. Expression of the Ocimum basilicum (sweet basil) geraniol synthase (GES) gene in a Saccharomyces cerevisiae wine strain substantially changed the terpene profile of wine produced from a non-aromatic grape variety. Under microvinification conditions, and without compromising other fermentative traits, the recombinant yeast excreted geraniol de novo at an amount (~750 μg/L) well exceeding (>10-fold) its threshold for olfactory perception and also exceeding the quantities present in wines obtained from highly aromatic Muscat grapes. Interestingly, geraniol was further metabolized by yeast enzymes to additional monoterpenes and esters: citronellol, linalool, nerol, citronellyl acetate and geranyl acetate, resulting in a total monoterpene concentration (~1,558 μg/L) 230-fold greater than that of the control. We also found that monoterpene profiles of wines derived from mixed fermentations were found to be determined by the composition of the initial yeast inocula suggesting the feasibility of producing 'à la carte' wines having predetermined monoterpene contents. Geraniol synthase-engineered yeasts demonstrate potential in the development of monoterpene enhanced wines.

  15. An integrated continuous improvement model of TPM, TPS and TQM for boosting profitability of manufacturing industries: An innovative model & guideline

    Directory of Open Access Journals (Sweden)

    Haftu Hailu

    2018-01-01

    Full Text Available The purpose of this research is to develop an integrated literature based TPM, TPS and TQM mod-el through PDCA cycle, and implementation guideline for the implementation of the model. At this time very few studies are available on this research area, even this research on integrated model of TPM, TPS and TQM practices, and implementation guideline are not corresponding. The method-ology to develop the model and the implementation guideline is based on identifying the uniqueness and common practices of TPM, TPS and TQM systems, existing practice of the integration and implementation guideline, identifying the existing gaps on the model and implementation guideline, developing new integrated TPM, TPS and TQM practice model, and implementation guideline. Previous very few studies of uniqueness, common practices and implementation guideline of the three systems are preserved. The findings of this research, an integrated cutting-edge model of TPM, TPS and TQM practices and implementation guidelines are developed. The originality / value of the developed model and implementation guideline enable manufacturing industries continuously to be competitive and profitable.

  16. Can contact allergy to p-phenylenediamine explain the high rates of terpene hydroperoxide allergy?

    DEFF Research Database (Denmark)

    Bennike, Niels Højsager; Lepoittevin, Jean Pierre; Johansen, Jeanne D.

    2017-01-01

    Background: Contact allergy to linalool hydroperoxides (Lin-OOHs) and limonene hydroperoxides (Lim-OOHs) is common. Similarly to what occurs with the terpene hydroperoxides, reactive intermediates formed from p-phenylenediamine (PPD) can cause oxidative modifications of tryptophan residues...... on proteins in mechanistic studies. Objectives: To test the hypothesis that patients sensitized to PPD are at increased risk of concomitant reactivity to either of the terpene hydroperoxides, owing to a ‘common pathway’ of skin protein oxidation. Methods: A database study of consecutively patch tested eczema...... patients (n = 3843) from 2012 to 2015, tested concomitantly with PPD, Lim-OOHs and Lin-OOHs, was performed. Associations were examined by level of concordance and odds ratios (ORs) adjusted for age, sex, and contact allergy to fragrance mix I and fragrance mix II. Results: Concomitant reactions to PPD were...

  17. Antioxidant, antibacterial and cytotoxic potential of silver nanoparticles synthesized using terpenes rich extract of Lantana camara L. leaves

    Directory of Open Access Journals (Sweden)

    Patil Shriniwas P.

    2017-07-01

    Full Text Available Several attempts have been made for green synthesis of silver nanoparticles (AgNPs using different plant extracts. Present study revealed that, antioxidant, antibacterial and cytotoxic AgNPs were synthesized using terpenes-rich extract (TRE of environmentally notorious Lantana camara L. leaves. AgNPs were characterized by advanced techniques like UV–Visible and Infra red spectroscopy; XRD, SEM techniques as terpenes coated sphere shaped NPs with average diameter 425 nm. Further, on evaluation, AgNPs were found to exhibit dose – dependent antioxidant potential, good to moderate antibacterial activity against Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa; and toxicity on Brine shrimp (A. salinanauplii with LD50 value 514.50 µg/ml.

  18. Impact of glutathione-enriched inactive dry yeast preparations on the stability of terpenes during model wine aging

    OpenAIRE

    Rodríguez-Bencomo, Juan José; Andújar-Ortiz, Inmaculada; Moreno-Arribas, M. Victoria; Simó, Carolina; González, Javier; Chana, Antonio; Dávalos, J.Z.; Pozo-Bayón, Mª Ángeles

    2014-01-01

    The impact of the addition of glutathione-enriched Inactive dry yeast preparations (g-IDYs) on the stability of some typical wine terpenes (linalool, α-terpineol, β-citronellol, and nerol) stored under accelerated oxidative conditions was evaluated in model wines. Additionally, the effects of a second type of IDY preparation with a different claim (fermentative nutrient) and the sole addition of commercial glutathione into the model wines were also assessed. Model wines were spiked with the l...

  19. QCM-Arrays for Sensing Terpenes in Fresh and Dried Herbs via Bio-Mimetic MIP Layers

    Directory of Open Access Journals (Sweden)

    Naseer Iqbal

    2010-06-01

    Full Text Available A piezoelectric 10 MHz multichannel quartz crystal microbalance (MQCM, coated with six molecularly imprinted polystyrene artificial recognition membranes have been developed for selective quantification of terpenes emanated from fresh and dried Lamiaceae family species, i.e., rosemary (Rosmarinus Officinalis L., basil (Ocimum Basilicum and sage (Salvia Officinalis. Optimal e-nose parameters, such as layer heights (1–6 KHz, sensitivity

  20. Chemodiversity of a Scots pine stand and implications for terpene air concentrations

    Science.gov (United States)

    Bäck, J.; Aalto, J.; Henriksson, M.; Hakola, H.; He, Q.; Boy, M.

    2012-02-01

    Atmospheric chemistry in background areas is strongly influenced by natural vegetation. Coniferous forests are known to produce large quantities of volatile vapors, especially terpenes. These compounds are reactive in the atmosphere, and contribute to the formation and growth of atmospheric new particles. Our aim was to analyze the variability of mono- and sesquiterpene emissions between Scots pine trees, in order to clarify the potential errors caused by using emission data obtained from only a few trees in atmospheric chemistry models. We also aimed at testing if stand history and seed origin has an influence on the chemotypic diversity. The inherited, chemotypic variability in mono- and sesquiterpene emission was studied in a seemingly homogeneous 48 yr-old stand in Southern Finland, where two areas differing in their stand regeneration history could be distinguished. Sampling was conducted in August 2009. Terpene concentrations in the air had been measured at the same site for seven years prior to branch sampling for chemotypes. Two main compounds, α-pinene and Δ3-carene formed together 40-97% of the monoterpene proportions in both the branch emissions and in the air concentrations. The data showed a bimodal distribution in emission composition, in particular in Δ3-carene emission within the studied population. 10% of the trees emitted mainly α-pinene and no Δ3-carene at all, whereas 20% of the trees where characterized as high Δ3-carene emitters (Δ3-carene forming >80% of total emitted monoterpene spectrum). An intermediate group of trees emitted equal amounts of both α-pinene and Δ3-carene. The emission pattern of trees at the area established using seeding as the artificial regeneration method differed from the naturally regenerated or planted trees, being mainly high Δ3-carene emitters. Some differences were also seen in e.g. camphene and limonene emissions between chemotypes, but sesquiterpene emissions did not differ significantly between trees

  1. The cellulose synthase companion proteins act non-redundantly with CELLULOSE SYNTHASE INTERACTING1/POM2 and CELLULOSE SYNTHASE 6

    OpenAIRE

    Endler, Anne; Schneider, Rene; Kesten, Christopher; Lampugnani, Edwin R.; Persson, Staffan

    2016-01-01

    Cellulose is a cell wall constituent that is essential for plant growth and development, and an important raw material for a range of industrial applications. Cellulose is synthesized at the plasma membrane by massive cellulose synthase (CesA) complexes that track along cortical microtubules in elongating cells of Arabidopsis through the activity of the protein CELLULOSE SYNTHASE INTERACTING1 (CSI1). In a recent study we identified another family of proteins that also are associated with the ...

  2. Impact of glutathione-enriched inactive dry yeast preparations on the stability of terpenes during model wine aging.

    Science.gov (United States)

    Rodríguez-Bencomo, Juan José; Andújar-Ortiz, Inmaculada; Moreno-Arribas, M Victoria; Simó, Carolina; González, Javier; Chana, Antonio; Dávalos, Juan; Pozo-Bayón, M Ángeles

    2014-02-12

    The impact of the addition of glutathione-enriched Inactive dry yeast preparations (g-IDYs) on the stability of some typical wine terpenes (linalool, α-terpineol, β-citronellol, and nerol) stored under accelerated oxidative conditions was evaluated in model wines. Additionally, the effects of a second type of IDY preparation with a different claim (fermentative nutrient) and the sole addition of commercial glutathione into the model wines were also assessed. Model wines were spiked with the low molecular weight fraction (loss of typical wine terpenes in model wines submitted to accelerated aging conditions. The g-IDY preparation did indeed release reduced GSH into the model wines, although this compound did not seem exclusively related to the protective effect on some aroma compounds determined in both model wines. The presence of other sulfur-containing compounds from yeast origin in g-IDY, and also the presence of small yeast peptides, such as methionine/tryptophan/tyrosine-containing tripeptide in both types of IDYs, seemed to be related to the antioxidant activity determined in the two permeates and to the minor loss of some terpenes in the model wines spiked with them.

  3. The effect of bleaching on the terpene chemistry of Plexaurella fusifera: evidence that zooxanthellae are not responsible for sesquiterpene production.

    Science.gov (United States)

    Frenz-Ross, Jamie L; Enticknap, Julie J; Kerr, Russell G

    2008-01-01

    The close association between marine invertebrates, zooxanthellae, and numerous bacteria gives rise to the question of the identity of the actual producer of secondary metabolites. In fall of 2005, a widespread bleaching event occurred throughout the Caribbean Sea in which some colonies of the gorgonian coral Plexaurella fusifera bleached. This study investigated whether zooxanthellae play a key role in the biosynthesis of secondary metabolite terpenes from P. fusifera. The extent of bleaching was examined by chlorophyll A analysis and also by zooxanthellae isolation and cell counting. The bleached and unbleached colonies were found to contain similar concentrations of eremophilene as the major terpene, and both exhibited similar biosynthetic capability as evaluated by the transformation of [C(1)-(3)H]-farnesyl diphosphate to the sesquiterpenes. Differences in bacterial communities between the bleached and unbleached colonies were analyzed using molecular techniques, and preliminary indications are that unbleached and bleached corals are dominated by low G + C firmicutes and gammaproteobacteria, respectively. It therefore appears that terpene biosynthesis can proceed independently of the zooxanthellae in P. fusifera, suggesting that the coral or a bacterium is the biosynthetic source.

  4. Effects of seasonal changes in feeding management under part-time grazing on terpene concentrations of ewes' milk.

    Science.gov (United States)

    Abilleira, Eunate; Virto, Mailo; Nájera, Ana Isabel; Albisu, Marta; Pérez-Elortondo, Francisco José; Ruiz de Gordoa, Juan Carlos; de Renobales, Mertxe; Barron, Luis Javier R

    2011-05-01

    Terpene composition of ewes' raw milk from nine commercial flocks was analysed from February to July. Ewes' diet consisted of concentrate and conserved forage in winter (indoor feeding) and part-time grazing from spring (transition and outdoor feeding). Regardless of the feeding, limonene and β-phellandrene were the most abundant monoterpenes and β-caryophyllene showed the highest concentrations among sesquiterpenes. Terpene content increased in the milks of commercial flocks when animals were reared under grazing management. Monoterpenes were detected in the milks of all the commercial flocks throughout the season, whereas sesquiterpenes were only detected in the milks from flocks grazing on non-cultivated community-owned grasslands in which a higher biodiversity of plant species grew. These preliminary results indicated that β-caryophyllene could be a potential pasture-diet marker in the case of milks from animals grazing a higher biodiversity of plant species but in-depth studies including information on terpene composition of plants ingested by the animals are necessary to evaluate the suitability of β-caryophyllene or another terpenoid compound as pasture biomarker.

  5. Think Pair Share (TPS as Method to Improve Student’s Learning Motivation and Learning Achievement

    Directory of Open Access Journals (Sweden)

    Hetika Hetika

    2018-03-01

    Full Text Available This research aims to find out the application of Think Pair Share (TPS learning method in improving learning motivation and learning achievement in the subject of Introduction to Accounting I of the Accounting Study Program students of Politeknik Harapan Bersama. The Method of data collection in this study used observation method, test method, and documentation method. The research instruments used observation sheet, questionnaire and test question. This research used Class Action Research Design which is an action implementation oriented research, with the aim of improving quality or problem solving in a group by carefully and observing the success rate due to the action. The method of analysis used descriptive qualitative and quantitative analysis method. The results showed that the application of Think Pair Share Learning (TPS Method can improve the Learning Motivation and Achievement. Before the implementation of the action, the obtained score is 67% then in the first cycle increases to 72%, and in the second cycle increasws to 80%. In addition, based on questionnaires distributed to students, it also increases the score of Accounting Learning Motivation where the score in the first cycle of 76% increases to 79%. In addition, in the first cycle, the score of pre test and post test of the students has increased from 68.86 to 76.71 while in the second cycle the score of pre test and post test of students has increased from 79.86 to 84.86.

  6. Application of GI-TPS Model to Skills Critical Thinking and Self Efficacy

    Directory of Open Access Journals (Sweden)

    Rusmansyah

    2017-12-01

    Full Text Available This research aims to know the: (1 the difference in critical thinking ability of students (2 difference in the results of learning (cognitive, affective, psychomotor (3 self efficacy of students (4 student response towards the learning model GI-TPS (Guided Inquiry-Think Pair Share on the hydrolysis of salt. This research use the pretest-posttest design nonequivalent control group design. A sample of 50 research students, namely class XI IPA 2 and XI IPA 3. Samples taken with purposive sampling technique. Engineering data collection using the test the critical thinking ability, cognitive learning results, observations, and the now. Analytical techniques descriptive analyses using data and analysis inferensial (test, test the normality of its homogeneity, and t-test. The results of this study suggest that (1 there is a difference significant critical thinking abilities (2 there is a difference in learning outcomes (cognitive, affective, psychomotor a significant (3 students who have self efficacy. (4 the students gave positive response towards the learning model GI-TPS material on the hydrolysis of salt.

  7. Identification and Characterization of the Iridoid Synthase Involved in Oleuropein Biosynthesis in Olive (Olea europaea) Fruits*

    Science.gov (United States)

    Kries, Hajo; Panara, Francesco; Baldoni, Luciana; O'Connor, Sarah E.; Osbourn, Anne

    2016-01-01

    The secoiridoids are the main class of specialized metabolites present in olive (Olea europaea L.) fruit. In particular, the secoiridoid oleuropein strongly influences olive oil quality because of its bitterness, which is a desirable trait. In addition, oleuropein possesses a wide range of pharmacological properties, including antioxidant, anti-inflammatory, and anti-cancer activities. In accordance, obtaining high oleuropein varieties is a main goal of molecular breeding programs. Here we use a transcriptomic approach to identify candidate genes belonging to the secoiridoid pathway in olive. From these candidates, we have functionally characterized the olive homologue of iridoid synthase (OeISY), an unusual terpene cyclase that couples an NAD (P)H-dependent 1,4-reduction step with a subsequent cyclization, and we provide evidence that OeISY likely generates the monoterpene scaffold of oleuropein in olive fruits. OeISY, the first pathway gene characterized for this type of secoiridoid, is a potential target for breeding programs in a high value secoiridoid-accumulating species. PMID:26709230

  8. Identification and Characterization of the Iridoid Synthase Involved in Oleuropein Biosynthesis in Olive (Olea europaea) Fruits.

    Science.gov (United States)

    Alagna, Fiammetta; Geu-Flores, Fernando; Kries, Hajo; Panara, Francesco; Baldoni, Luciana; O'Connor, Sarah E; Osbourn, Anne

    2016-03-11

    The secoiridoids are the main class of specialized metabolites present in olive (Olea europaea L.) fruit. In particular, the secoiridoid oleuropein strongly influences olive oil quality because of its bitterness, which is a desirable trait. In addition, oleuropein possesses a wide range of pharmacological properties, including antioxidant, anti-inflammatory, and anti-cancer activities. In accordance, obtaining high oleuropein varieties is a main goal of molecular breeding programs. Here we use a transcriptomic approach to identify candidate genes belonging to the secoiridoid pathway in olive. From these candidates, we have functionally characterized the olive homologue of iridoid synthase (OeISY), an unusual terpene cyclase that couples an NAD (P)H-dependent 1,4-reduction step with a subsequent cyclization, and we provide evidence that OeISY likely generates the monoterpene scaffold of oleuropein in olive fruits. OeISY, the first pathway gene characterized for this type of secoiridoid, is a potential target for breeding programs in a high value secoiridoid-accumulating species. © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

  9. Oil production at different stages of leaf development in Lippia alba

    Directory of Open Access Journals (Sweden)

    Diego Pandeló

    2012-06-01

    Full Text Available The aim of this work was to analyze terpene oil production and terpene synthases (TPS gene expression from leaves at different developmental stages of different chemotypes of Lippia alba (Mill. N.E. Br. ex Britton & P. Wilson, Verbenaceae. Hydro-distilled essential oil were used for chemical analysis and gene expression of three monoterpene synthase genes called LaTPS12, LaTPS23 and LaTPS25 were used for analyses of gene expression associated to oil production. The putative genes were associated to TPS-b gene class. Semi-quantitative PCR and quantitative PCR (qPCR analysis were used to investigate the expression profile of those three putative genes in different leaf stages and different chemotypes. Additionally, total oil production and gene expression of putative TPS genes cloned from L. alba chemotype linalool were evaluated at different stages of leaf development. The expression level of those three genes was higher when the highest oil production was observed, mainly in young leaves at the fourth nodal segment for all evaluated chemotypes. Total oil production was higher at leaves that had unopened trichomes. We also observed that the 1mM of MeJA treatment increased the gene expression in all chemotypes after 24 h elicitation.

  10. Oil production at different stages of leaf development in Lippia alba

    Directory of Open Access Journals (Sweden)

    Diego Pandeló

    2012-01-01

    Full Text Available The aim of this work was to analyze terpene oil production and terpene synthases (TPS gene expression from leaves at different developmental stages of different chemotypes of Lippia alba (Mill. N.E. Br. ex Britton & P. Wilson, Verbenaceae. Hydro-distilled essential oil were used for chemical analysis and gene expression of three monoterpene synthase genes called LaTPS12, LaTPS23 and LaTPS25 were used for analyses of gene expression associated to oil production. The putative genes were associated to TPS-b gene class. Semi-quantitative PCR and quantitative PCR (qPCR analysis were used to investigate the expression profile of those three putative genes in different leaf stages and different chemotypes. Additionally, total oil production and gene expression of putative TPS genes cloned from L. alba chemotype linalool were evaluated at different stages of leaf development. The expression level of those three genes was higher when the highest oil production was observed, mainly in young leaves at the fourth nodal segment for all evaluated chemotypes. Total oil production was higher at leaves that had unopened trichomes. We also observed that the 1mM of MeJA treatment increased the gene expression in all chemotypes after 24 h elicitation.

  11. Pragmatic evaluation of the Toyota Production System (TPS analysis procedure for problem solving with entry-level nurses

    Directory of Open Access Journals (Sweden)

    Lukasz Maciej Mazur

    2008-12-01

    Full Text Available Medication errors occurring in hospitals are a growing national concern. These medication errors and their related costs (or wastes are seen as major factors leading to increased patient safety risks and increased waste in the hospital setting.  This article presents a study in which sixteen entry-level nurses utilized a Toyota Production System (TPS analysis procedure to solve medication delivery problems at one community hospital. The objective of this research was to study and evaluate the TPS analysis procedure for problem solving with entry-level nurses. Personal journals, focus group discussions, and a survey study were used to collect data about entry-level nurses’ perceptions of using the TPS problem solving approach to study medication delivery. A regression analysis was used to identify characteristics that enhance problem solving efforts. In addition, propositions for effective problem solving by entry-level nurses to aid in the reduction of medication errors in healthcare delivery settings are offered.

  12. Endothelial nitric oxide synthase gene polymorphisms associated ...

    African Journals Online (AJOL)

    STORAGESEVER

    2010-05-24

    May 24, 2010 ... chronic periodontitis (CP), 31 with gingivitis (G) and 50 healthy controls. Probing depth ..... Periodontal disease in pregnancy I. Prevalence and severity. ... endothelial nitric oxide synthase gene in premenopausal women with.

  13. Secondary organic aerosol from ozone-initiated reactions with terpene-rich household products

    Energy Technology Data Exchange (ETDEWEB)

    Coleman, Beverly; Coleman, Beverly K.; Lunden, Melissa M.; Destaillats, Hugo; Nazaroff, William W.

    2008-01-01

    We analyzed secondary organic aerosol (SOA) data from a series of small-chamber experiments in which terpene-rich vapors from household products were combined with ozone under conditions analogous to product use indoors. Reagents were introduced into a continuously ventilated 198 L chamber at steady rates. Consistently, at the time of ozone introduction, nucleation occurred exhibiting behavior similar to atmospheric events. The initial nucleation burst and growth was followed by a period in which approximately stable particle levels were established reflecting a balance between new particle formation, condensational growth, and removal by ventilation. Airborne particles were measured with a scanning mobility particle sizer (SMPS, 10 to 400 nm) in every experiment and with an optical particle counter (OPC, 0.1 to 2.0 ?m) in a subset. Parameters for a three-mode lognormal fit to the size distribution at steady state were determined for each experiment. Increasing the supply ozone level increased the steady-state mass concentration and yield of SOA from each product tested. Decreasing the air-exchange rate increased the yield. The steady-state fine-particle mass concentration (PM1.1) ranged from 10 to> 300 mu g m-3 and yields ranged from 5percent to 37percent. Steady-state nucleation rates and SOA mass formation rates were on the order of 10 cm-3 s-1 and 10 mu g m-3 min-1, respectively.

  14. Terpenes, Phenylpropanoids, Sulfur and Other Essential Oil Constituents as Inhibitors of Cholinesterases.

    Science.gov (United States)

    Burcul, Franko; Blazevic, Ivica; Radan, Mila; Politeo, Olivera

    2018-03-29

    Essential oils constituents are a diverse family of low molecular weight organic compounds with comprehensive biological activity. According to their chemical structure these active compounds can be divided into four major groups: terpenes, terpenoids, phenylpropenes, and "others". In addition, they may contain diverse functional groups according to which they can be classified as hydrocarbons (monoterpenes, sesquiterpenes, and aliphatic hydrocarbons); oxygenated compounds (monoterpene and sesquiterpene alcohols, aldehydes, ketones, esters, and other oxygenated compounds); and sulfur and/or nitrogen sulfur-containing compounds (thioesters, sulfides, isothiocyantes, nitriles, and others). Compounds that act as cholinesterase inhibitors still represent the only pharmacological treatment of Alzheimer´s disease. Numerous in vitro studies showed that some compounds, found in essential oils, have a promising cholinesterase inhibitory activity, such as α-pinene, δ-3-carene, 1,8-cineole, carvacrol, thymohydroquinone, α- and β-asarone, anethole, etc. This review summarizes the most relevant research published to date on essential oil constituents and their acetylcholinesterase/butyrylcholinesterase inhibitory potential as well as their structure related activity, synergistic and antagonistic effects. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

  15. [Regulation of terpene metabolism]. Annual progress report, March 15, 1990--March 14, 1991

    Energy Technology Data Exchange (ETDEWEB)

    Croteau, R.

    1991-12-31

    During the last grant period, we have completed studies on the key pathways of monoterpene biosynthesis and catabolism in sage and peppermint, and have, by several lines of evidence, deciphered the rate-limiting step of each pathway. We have at least partially purified and characterized the relevant enzymes of each pathway. We have made a strong case, based on analytical, in vivo, and in vitro studies, that terpene accumulation depends upon the balance between biosynthesis and catabolism, and provided supporting evidence that these processes are developmentally-regulated and very closely associated with senescence of the oil glands. Oil gland ontogeny has been characterized at the ultrastructural level. We have exploited foliar-applied bioregulators to delay gland senescence, and have developed tissue explant and cell culture systems to study several elusive aspects of catabolism. We have isolated pure gland cell clusters and localized monoterpene biosynthesis and catabolism within these structures, and have used these preparations as starting materials for the purification to homogeneity of target ``regulatory`` enzymes. We have thus developed the necessary background knowledge, based on a firm understanding of enzymology, as well as the necessary experimental tools for studying the regulation of monoterpene metabolism at the molecular level. Furthermore, we are now in a position to extend our systematic approach to other terpenoid classes (C{sub 15}-C{sub 30}) produced by oil glands.

  16. Oxidized limonene and oxidized linalool - concomitant contact allergy to common fragrance terpenes.

    Science.gov (United States)

    Bråred Christensson, Johanna; Karlberg, Ann-Therese; Andersen, Klaus E; Bruze, Magnus; Johansen, Jeanne D; Garcia-Bravo, Begoña; Giménez Arnau, Ana; Goh, Chee-Leok; Nixon, Rosemary; White, Ian R

    2016-05-01

    Limonene and linalool are common fragrance terpenes. Both oxidized R-limonene and oxidized linalool have recently been patch tested in an international setting, showing contact allergy in 5.2% and 6.9% of dermatitis patients, respectively. To investigate concomitant reactions between oxidized R-limonene and oxidized linalool in consecutive dermatitis patients. Oxidized R-limonene 3.0% (containing limonene hydroperoxides 0.33%) and oxidized linalool 6% (linalool hydroperoxides 1%) in petrolatum were tested in 2900 consecutive dermatitis patients in Australia, Denmark, Singapore, Spain, Sweden, and the United Kingdom. A total of 281 patients reacted to either oxidized R-limonene or oxidized linalool. Of these, 25% had concomitant reactions to both compounds, whereas 29% reacted only to oxidized R-limonene and 46% only to oxidized linalool. Of the 152 patients reacting to oxidized R-limonene, 46% reacted to oxidized linalool, whereas 35% of the 200 patients reacting to oxidized linalool also reacted to oxidized R-limonene. The majority of the patients (75%) reacted to only one of the oxidation mixtures, thus supporting the specificity of the reactions. The concomitant reactions to the two fragrance allergens suggest multiple sensitizations, which most likely reflect the exposure to the different fragrance materials in various types of consumer products. This is in accordance with what is generally seen for patch test reactions to fragrance materials. © 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  17. Keefektifan setting TPS dalam pendekatan discovery learning dan problem-based learning pada pembelajaran materi lingkaran SMP

    Directory of Open Access Journals (Sweden)

    Rahmi Hidayati

    2017-05-01

    The purpose of this study was to describe the effectiveness of setting Think Pair Share (TPS in the approach to discovery learning and problem-based learning in terms of student achievement, mathematical communication skills, and interpersonal skills of the student.  This study was a quasi-experimental study using the pretest-posttest nonequivalent group design. The research population comprised all Year VIII students of SMP Negeri 1 Yogyakarta. The research sample was randomly selected from eight classes, two classes were elected. The instrument used in this study is the learning achievement test, a test of mathematical communication skills, and interpersonal skills student questionnaires. To test the effectiveness of setting Think Pair Share (TPS in the approach to discovery learning and problem-based learning, the one sample t-test was carried out. Then, to investigate the difference in effectiveness between the setting Think Pair Share (TPS in the approach to discovery learning and problem-based learning, the Multivariate Analysis of Variance (MANOVA was carried out. The research findings indicate that the setting TPS discovery approach to learning and problem-based approach to learning (PBL is effective in terms of learning achievement, mathematical communication skills, and interpersonal skills of the students. No difference in effectiveness between setting TPS discovery approach to learning and problem-based learning (PBL in terms of learning achievement, mathematical communication skills, and interpersonal skills of the students. Keywords: TPS setting in discovery learning approach, in problem-based learning, academic achievement, mathematical communication skills, and interpersonal skills of the student

  18. Analysis on TOYOTA's Production Mode TPS%解读丰田生产模式(TPS)--走近丰田叉车系列报道之三

    Institute of Scientific and Technical Information of China (English)

    陈方建

    2004-01-01

    @@ 丰田生产模式(Toyota Production System,TPS)对国内读者来说并不陌生,它产生于日本丰田公司,是"为实现企业对员工、社会和产品负责的目的,以彻底杜绝浪费的思想为目标,在连续改善的基础上,采用准时化与自动化方式与方法,追求制造产品合理性的一种生产方式".通过此次赴日对丰田叉车日本总部的参观考察,笔者对这一在全世界广受瞩目的生产模式有了更进一步的了解与认识,在此愿提出来与大家共享.

  19. Structure of the ent -Copalyl Diphosphate Synthase PtmT2 from Streptomyces platensis CB00739, a Bacterial Type II Diterpene Synthase

    Energy Technology Data Exchange (ETDEWEB)

    Rudolf, Jeffrey D.; Dong, Liao-Bin; Cao, Hongnan; Hatzos-Skintges, Catherine; Osipiuk, Jerzy; Endres, Michael; Chang, Chin-Yuan; Ma, Ming; Babnigg, Gyorgy; Joachimiak, Andrzej; Phillips, George N.; Shen, Ben

    2016-08-31

    Terpenoids are the largest and most structurally diverse family of natural products found in nature, yet their presence in bacteria is underappreciated. The carbon skeletons of terpenoids are generated through carbocation-dependent cyclization cascades catalyzed by terpene synthases (TSs). Type I and type II TSs initiate cyclization via diphosphate ionization and protonation, respectively, and protein structures of both types are known. Most plant diterpene synthases (DTSs) possess three alpha-helical domains (alpha beta gamma), which are thought to have arisen from the fusion of discrete, ancestral bacterial type I TSs (alpha) and type II TSs (beta gamma). Type II DTSs of bacterial origin, of which there are no structurally characterized members, are a missing piece in the structural evolution of TSs. Here, we report the first crystal structure of a type II DTS from bacteria. PtnaT2 from Streptomyces platensis CB00739 was verified as an ent-copalyl diphosphate synthase involved in the biosynthesis of platensimycin and platencin. The crystal structure of PtmT2 was solved at a resolution of 1.80 angstrom, and docking studies suggest the catalytically active conformation of geranylgeranyl diphosphate (GGPP). Site-directed mutagenesis confirmed residues involved in binding the diphosphate moiety of GGPP and identified DxxxxE as a potential Mg2+-binding motif for type II DTSs of bacterial origin. Finally, both the shape and physicochemical properties of the active sites are responsible for determining specific catalytic outcomes of TSs. The structure of PtmT2 fundamentally advances the knowledge of bacterial TSs, their mechanisms, and their role in the evolution of TSs.

  20. Heteromeric and homomeric geranyl diphosphate synthases from Catharanthus roseus and their role in monoterpene indole alkaloid biosynthesis.

    Science.gov (United States)

    Rai, Avanish; Smita, Shachi S; Singh, Anup Kumar; Shanker, Karuna; Nagegowda, Dinesh A

    2013-09-01

    Catharanthus roseus is the sole source of two most important monoterpene indole alkaloid (MIA) anti-cancer agents: vinblastine and vincristine. MIAs possess a terpene and an indole moiety derived from terpenoid and shikimate pathways, respectively. Geranyl diphosphate (GPP), the entry point to the formation of terpene moiety, is a product of the condensation of isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP) by GPP synthase (GPPS). Here, we report three genes encoding proteins with sequence similarity to large subunit (CrGPPS.LSU) and small subunit (CrGPPS.SSU) of heteromeric GPPSs, and a homomeric GPPSs. CrGPPS.LSU is a bifunctional enzyme producing both GPP and geranyl geranyl diphosphate (GGPP), CrGPPS.SSU is inactive, whereas CrGPPS is a homomeric enzyme forming GPP. Co-expression of both subunits in Escherichia coli resulted in heteromeric enzyme with enhanced activity producing only GPP. While CrGPPS.LSU and CrGPPS showed higher expression in older and younger leaves, respectively, CrGPPS.SSU showed an increasing trend and decreased gradually. Methyl jasmonate (MeJA) treatment of leaves significantly induced the expression of only CrGPPS.SSU. GFP localization indicated that CrGPPS.SSU is plastidial whereas CrGPPS is mitochondrial. Transient overexpression of AmGPPS.SSU in C. roseus leaves resulted in increased vindoline, immediate monomeric precursor of vinblastine and vincristine. Although C. roseus has both heteromeric and homomeric GPPS enzymes, our results implicate the involvement of only heteromeric GPPS with CrGPPS.SSU regulating the GPP flux for MIA biosynthesis.

  1. Overview of the 6 Meter HIAD Inflatable Structure and Flexible TPS Static Load Test Series

    Science.gov (United States)

    Swanson, Greg; Kazemba, Cole; Johnson, Keith; Calomino, Anthony; Hughes, Steve; Cassell, Alan; Cheatwood, Neil

    2014-01-01

    To support NASAs long term goal of landing humans on Mars, technologies which enable the landing of heavy payloads are being developed. Current entry, decent, and landing technologies are not practical for this class of payloads due to geometric constraints dictated by current launch vehicle fairing limitations. Therefore, past and present technologies are now being explored to provide a mass and volume efficient solution to atmospheric entry, including Hypersonic Inflatable Aerodynamic Decelerators (HIADs). At the beginning of 2014, a 6m HIAD inflatable structure with an integrated flexible thermal protection system (TPS) was subjected to a static load test series to verify the designs structural performance. The 6m HIAD structure was constructed in a stacked toroid configuration using nine inflatable torus segments composed of fiber reinforced thin films, which were joined together using adhesives and high strength textile woven structural straps to help distribute the loads throughout the inflatable structure. The 6m flexible TPS was constructed using multiple layers of high performance materials to protect the inflatable structure from heat loads that would be seen during atmospheric entry. To perform the static load test series, a custom test fixture was constructed. The fixture consisted of a structural tub rim with enough height to allow for displacement of the inflatable structure as loads were applied. The bottom of the tub rim had an airtight seal with the floor. The centerbody of the inflatable structure was attached to a pedestal mount as seen in Figure 1. Using an impermeable membrane seal draped over the test article, partial vacuum was pulled beneath the HIAD, resulting in a uniform static pressure load applied to the outer surface. During the test series an extensive amount of instrumentation was used to provide many data sets including: deformed shape, shoulder deflection, strap loads, cord loads, inflation pressures, and applied static load

  2. Design and fabrication of the vacuum systems for TPS pulsed septum magnets

    Science.gov (United States)

    Chan, C. K.; Chang, C. C.; Chen, C. L.; Yang, C. S.; Chen, C. S.; Lin, F. Y.; Chen, J. R.

    2014-11-01

    Three in-air pulsed septum magnets were developed to inject and extract electron beams for the 3 GeV synchrotron facility, the Taiwan Photon Source (TPS). The vacuum chamber is a novel combined aluminium-stainless steel design, using a bimetallic flange to connect the two material types. To evaluate the vacuum performances of these vacuum chambers, we set up a test bench at which we simultaneously measure the rates of thermal outgassing of the aluminium chamber and the septum tube with a throughput method. The test result indicates that the rate q72 of thermal outgassing measured after 1 day from baking at 150 °C was 1×10-13 mbar L s-1 cm-2. The magnetic leakage measurements show the combination of conductor slitting, magnetic shielding and the aluminium vacuum chamber reduce the peak value of the leakage field integral to ~10 G cm along the trajectory of the stored beam.

  3. Conformal Ablative Thermal Protection Systems (CA-TPS) for Venus and Saturn Backshells

    Science.gov (United States)

    Beck, R.; Gasch, M.; Stackpoole, M.; Wilder, M.; Boghozian, T.; Chavez-Garcia, J.; Prabhu, D.; Kazemba, C.; Venkatapathy, E.

    2015-01-01

    The new conformal ablator C-PICA, which was developed under STMD GCD, is an optimal candidate for use on the backshells for high velocity entry vehicles at both Venus and Saturn. The material has been tested at heat fluxes up to 400 Wcm2 in shear and over 1800 Wcm2 and 1.5 atm in stagnation with good results. C-PICA has similar density to PICA, but shows half the thermal penetration and similar recession at the same conditions, allowing for a lighter weight TPS to be flown. This poster for VEXAG will show the progress made in the development of the material and why it should be considered for use.

  4. Installation and commissioning of a cryogen distribution system for the TPS project

    Science.gov (United States)

    Tsai, H. H.; Hsiao, F. Z.; Li, H. C.; Lin, M. C.; Wang, C.; Liao, W. R.; Lin, T. F.; Chiou, W. S.; Chang, S. H.; Chuang, P. S. D.

    2016-07-01

    A cryogen distribution system was installed and commissioned to transfer liquid nitrogen (LN2) and liquid helium (LHe) from storage dewars to superconducting radio-frequency (SRF) cavities for the 3-GeV Taiwan Photon Source (TPS) project. The cryogen distribution system comprises one distribution valve box (DVB), four control valve boxes (CVB) and seven sections of multichannel transfer line (MCL). The DVB distributes the LHe and LN2 to the CVB, and then to the SRF cavities through independent vacuum-jacketed transfer lines. The vaporized GHe and GN2 from the cryomodules are collected via the MCL. The cryogen distribution system was installed and commissioned from October 2014 to the end of March 2015. This paper presents the installation, pre-commissioning and commissioning of the cryogen distribution system, and describes the heat load test. Thermal acoustic oscillation (TAO) was found in the GHe process line; this phenomenon and its solution are also presented and discussed.

  5. Numerical Simulation and Mechanical Design for TPS Electron Beam Position Monitors

    Science.gov (United States)

    Hsueh, H. P.; Kuan, C. K.; Ueng, T. S.; Hsiung, G. Y.; Chen, J. R.

    2007-01-01

    Comprehensive study on the mechanical design and numerical simulation for the high resolution electron beam position monitors are key steps to build the newly proposed 3rd generation synchrotron radiation research facility, Taiwan Photon Source (TPS). With more advanced electromagnetic simulation tool like MAFIA tailored specifically for particle accelerator, the design for the high resolution electron beam position monitors can be tested in such environment before they are experimentally tested. The design goal of our high resolution electron beam position monitors is to get the best resolution through sensitivity and signal optimization. The definitions and differences between resolution and sensitivity of electron beam position monitors will be explained. The design consideration is also explained. Prototype deign has been carried out and the related simulations were also carried out with MAFIA. The results are presented here. Sensitivity as high as 200 in x direction has been achieved in x direction at 500 MHz.

  6. Numerical Simulation and Mechanical Design for TPS Electron Beam Position Monitors

    International Nuclear Information System (INIS)

    Hsueh, H. P.; Kuan, C. K.; Ueng, T. S.; Hsiung, G. Y.; Chen, J. R.

    2007-01-01

    Comprehensive study on the mechanical design and numerical simulation for the high resolution electron beam position monitors are key steps to build the newly proposed 3rd generation synchrotron radiation research facility, Taiwan Photon Source (TPS). With more advanced electromagnetic simulation tool like MAFIA tailored specifically for particle accelerator, the design for the high resolution electron beam position monitors can be tested in such environment before they are experimentally tested. The design goal of our high resolution electron beam position monitors is to get the best resolution through sensitivity and signal optimization. The definitions and differences between resolution and sensitivity of electron beam position monitors will be explained. The design consideration is also explained. Prototype deign has been carried out and the related simulations were also carried out with MAFIA. The results are presented here. Sensitivity as high as 200 in x direction has been achieved in x direction at 500 MHz

  7. Meningkatkan Prestasi Belajar IPA Melalui Model Pembelajaran Kooperatif Think-Pairs Share (TPS

    Directory of Open Access Journals (Sweden)

    Rusman Rusman

    2014-02-01

    Full Text Available Mata pelajaran IPA merupakan mata pelajaran yang menekankan siswa untuk mencari atau menemukan pengetahuanya sendiri. Model pembelajaran yang digunakan guru sangat berpengaruh dalam menciptakan situasi belajar yang benar-benar menyenangkan dan mendukung kelancaran proses belajar mengajar, serta sangat membantu dalam pencapaian prestasi belajar yang memuaskan. Akan tetapi, dalam pelaksanaan di lapangan pembelajaran banyak didominasi oleh guru sehingga kurang mampu membangun persepsi, minat, dan sikap siswa yang lebih baik. Hal tersebut juga dijumpai di SDN Pinggir Papas 1 Sumenep, siswa sulit memahami materi yang ada. Kendala yang terjadi adalah siswa merasa bosan dan tidak berminat mengikuti pembelajaran IPA. Hasil nilai ulangan harian dari 25 siswa hanya 9 siswa (36% yang mendapatkan nilai di atas KKM (65. Dari hasil tersebut, peneliti merasa perlu sekali melakukan perbaikan pembelajaran agar sehingga hasil belajar siswa dapat ditingkatkan, yaitu dengan menerapkan model  pembelajaran kooperatif Think-Pair-Share (TPS. Sesuai rumusan masalah “apakah model pembelajaran kooperatif Think-Pairs-Share (TPS dapat meningkatkan Prestasi Belajar IPA pada Siswa Kelas IIIA SDN Pinggir Papas 1 Sumenep?”, maka dilakukan metode penelitian dengan observasi dan tes. Hasil dari dua siklus penelitian yang melalui tahapan perencanaan, pelaksanaan, pengamatan, dan refleksi diperoleh hasil prestasi belajar siswa meningkat dari 67,8 dengan ketuntasan belajar 72% menjadi 80,4 dengan ketuntasan 100%. Sehingga dapat disimpulkan bahwa penerapan model pembelajaran kooperatif Think Pairs Share mampu meningkatkan prestasi belajar IPA pada siswa kelas IIIA SDN Pinggir Papas 1 Sumenep, dengan peningkatan yang sangat signifikan yaitu sebesar 28% melebihi kriteria peningkatan yang ditentukan yaitu 20%. Selain itu, model pembelajaran kooperatif Think Pairs Share efektif digunakan sebagai salah satu metode pembelajaran di kelas karena dapat menjadikan siswa aktif dalam

  8. Morphologies, Processing and Properties of Ceramic Foams and Their Potential as TPS Materials

    Science.gov (United States)

    Stackpoole, Mairead; Simoes, Conan R.; Johnson, Sylvia M.

    2002-01-01

    The current research is focused on processing ceramic foams with compositions that have potential as a thermal protection material. The use of pre-ceramic polymers with the addition of sacrificial blowing agents or sacrificial fillers offers a viable approach to form either open or closed cell insulation. Our work demonstrates that this is a feasible method to form refractory ceramic foams at relatively low processing temperatures. It is possible to foam complex shapes then pyrolize the system to form a ceramic while retaining the shape of the unfired foam. Initial work focused on identifying suitable pre-ceramic polymers with desired properties such as ceramic yield and chemical make up of the pyrolysis product after firing. We focused on making foams in the Si system (Sic, Si02, Si-0-C), which is in use in current acreage TPS systems. Ceramic foams with different architectures were formed from the pyrolysis of pre-ceramic polymers at 1200 C in different atmospheres. In some systems a sacrificial polyurethane was used as the blowing agent. We have also processed foams using sacrificial fillers to introduce controlled cell sizes. Each sacrificial filler or blowing agent leads to a unique morphology. The effect of different fillers on foam morphologies and the characterization of these foams in terms of mechanical and thermal properties are presented. We have conducted preliminary arc jet testing on selected foams with the materials being exposed to typical re-entry conditions for acreage TPS and these results will be discussed. Foams processed using these approaches have bulk densities ranging from 0.15 to 0.9 g/cm3 and cell sizes ranging from 5 to 500 pm. Compression strengths ranged from 2 to 7 MPa for these systems. Finally, preliminary oxidation studies have been conducted on selected systems and will be discussed.

  9. Tissue polypeptide-specific antigen (TPS) determinations before and during intermittent maximal androgen blockade in patients with metastatic prostatic carcinoma

    NARCIS (Netherlands)

    Kil, P. J. M.; Goldschmidt, H. M. J.; Wieggers, B. J. A.; Kariakine, O. B.; Studer, U. E.; Whelan, P.; Hetherington, J.; de Reijke, Th M.; Hoekstra, J. W.; Collette, L.

    2003-01-01

    To evaluate the prognostic significance of serially measured tissue polypeptide-specific antigen (TPS) levels in patients with metastatic prostatic carcinoma treated with intermittent maximal androgen blockade (MAB). To determine its value with respect to predicting response to treatment and time to

  10. "ELIP-MARC" Activities via TPS of Cooperative Learning to Improve Student's Mathematical Reasoning

    Science.gov (United States)

    Ulya, Wisulah Titah; Purwanto; Parta, I. Nengah; Mulyati, Sri

    2017-01-01

    The purpose of this study is to describe and generate interaction model of learning through "Elip-Marc" activity via "TPS" cooperative learning in order to improve student's mathematical reasoning who have valid, practical and effective criteria. "Elip-Marc" is an acronym of eliciting, inserting, pressing,…

  11. Adaptation mechanisms of bacteria during the degradation of polychlorinated biphenyls in the presence of natural and synthetic terpenes as potential degradation inducers

    Energy Technology Data Exchange (ETDEWEB)

    Zoradova-Murinova, Slavomira; Dudasova, Hana; Lukacova, Lucia; Certik, Milan; Dercova, Katarina [Slovak Univ. of Technology, Bratislava (Slovakia). Inst. of Biotechnology and Food Science; Silharova, Katarina; Vrana, Branislav [Water Research Institute, Bratislava (Slovakia)

    2012-06-15

    In this study, we examined the effect of polychlorinated biphenyls (PCBs) in the presence of natural and synthetic terpenes and biphenyl on biomass production, lipid accumulation, and membrane adaptation mechanisms of two PCB-degrading bacterial strains Pseudomonas stutzeri and Burkholderia xenovorans LB400. According to the results obtained, it could be concluded that natural terpenes, mainly those contained in ivy leaves and pine needles, decreased adaptation responses induced by PCBs in these strains. The adaptation processes under investigation included growth inhibition, lipid accumulation, composition of fatty acids, cis/trans isomerization, and membrane saturation. Growth inhibition effect decreased upon addition of these natural compounds to the medium. The amount of unsaturated fatty acids that can lead to elevated membrane fluidity increased in both strains after the addition of the two natural terpene sources. The cells adaptation changes were more prominent in the presence of carvone, limonene, and biphenyl than in the presence of natural terpenes, as indicated by growth inhibition, lipid accumulation, and cis/trans isomerization. Addition of biphenyl and carvone simultaneously with PCBs increased the trans/cis ratio of fatty acids in membrane fractions probably as a result of fluidizing effects of PCBs. This stimulation is more pronounced in the presence of PCBs as a sole carbon source. This suggests that PCBs alone have a stronger effect on bacterial membrane adaptation mechanisms than when added together with biphenyl or natural or synthetic terpenes. (orig.)

  12. Etude des melanges co-continus d'acide polylactique et d'amidon thermoplastique (PLA/TPS)

    Science.gov (United States)

    Chavez Garcia, Maria Graciela

    Les melanges co-continus sont des melanges polymeriques ou chaque composant se trouve dans une phase continue. Pour cette raison, les caracteristiques de chacun des composants se combinent et il en resulte un materiau avec une morphologie et des proprietes particulieres. L'acide polylactique (PLA) et l'amidon thermoplastique (TPS) sont des biopolymeres qui proviennent de ressources renouvelables et qui sont biodegradables. Dans ce projet, differents melanges de PLA et TPS a une haute concentration de TPS ont ete prepares dans une extrudeuse bi-vis afin de generer des structures co-continues. Grace a la technique de lixiviation selective, le TPS est enleve pour creer une structure poreuse de PLA qui a pu etre analysee au moyen de la microtomographie R-X et de la microscopie electronique a balayage MEB. L'analyse des images 2D et 3D confirme la presence de la structure co-continue dans les melanges dont la concentration en TPS. se situe entre 66% et 80%. L'effet de deux plastifiants, le glycerol seul et le melange de glycerol et de sorbitol, dans la formulation de TPS est etudie dans ce travail. De plus, nous avons evalue l'effet du PLA greffe a l'anhydride maleique (PLAg) en tant que compatibilisant. On a trouve que la phase de TPS obtenue avec le glycerol est plus grande. L'effet de recuit sur la taille de phases est aussi analyse. Grace aux memes techniques d'analyse, on a etudie l'effet du procede de moulage par injection sur la morphologie. On a constate que les pieces injectees presentent une microstructure heterogene et differente entre la surface et le centre de la piece. Pres de la surface, une peau plus riche en PLA est presente et les phases de TPS y sont allongees sous forme de lamelles. Plus au centre de la piece, une morphologie plus cellulaire est observee pour chaque phase continue. L'effet des formulations sur les proprietes mecaniques a aussi ete etudie. Les pieces injectees dont la concentration de TPS est plus grande presentent une moindre

  13. Terpene Profile, Leaf Anatomy, and Enzyme Activity of Resistant and Susceptible Cocoa Clonesto Vascular Streak Dieback Disease

    Directory of Open Access Journals (Sweden)

    Adi Prawoto

    2014-10-01

    Full Text Available Vascular-streak dieback (VSD, Oncobasidium theobromae is the most prevalent disease of Theobroma cacao L. in Indonesia. This study aims to analyze resistance mechanism to VSD based on terpene profile, leaf anatomy, chitinase, and peroxidase study. Resistant clones of Sulawesi 1 and Sca 6 and susceptible clones of ICS 60 and TSH 858 were used for terpene profile, leaf anatomy analysis, chitinase, peroxides, polyphenol, lignin, and cellulose analysis. Those clones and KEE 2, KKM 22 and ICS 13 were used for peroxides analysis. For trichome study, the resistant clones of Sulawesi 1, Sca 6, KEE 2, and KKM 22, and susceptible clones of ICS 60 and TSH 858 were used. GCMS analysis showed that chromatogram pattern of resistant and susceptible groups were quite similar, but resistant clones contained 22% more components than the susceptible ones. Resistant clones contained groups of pinene, decane, myrcene, and octadecanoic acid, while those substances on usceptible clones were absent. Trichome was thicker on younger leaf, and its density on the basal was higher than that on the middle and tip leaf parts. Trichome density of resistant clone was not always thicker than that of susceptible ones. On resistant clones, stomatal density was lower and width of stomate pits was narrower, while thickness of epidermis layer and pallisade parenchym were higher. Polyphenol content of resistant clones were higher but lignin and cellulose of both groups were similar. Chitinase activity which has a role in hydrolysis of mycelia cell wall was higher on the resistant clones, but peroxides which has a role in polymeration of lignin biosynthesis was similar between both groups. It is concluded that groups of terpene pinene, decane, myrcene, and octadecanoic acid, thickness of leaf epidermis, density and width of stomata pit, and chitinase activity plays important role in cocoa resistance to VSD. Key words: Theobroma cacaoL., clone, vascular-streak dieback, resistance, leaf

  14. Bacillus caldolyticus prs gene encoding phosphoribosyldiphosphate synthase

    DEFF Research Database (Denmark)

    Krath, Britta N.; Hove-Jensen, Bjarne

    1996-01-01

    The prs gene, encoding phosphoribosyl-diphosphate (PRPP) synthase, as well as the flanking DNA sequences were cloned and sequenced from the Gram-positive thermophile, Bacillus caldolyticus. Comparison with the homologous sequences from the mesophile, Bacillus subtilis, revealed a gene (gca......D) encoding N-acetylglucosamine-l-phosphate uridyltransferase upstream of prs, and a gene homologous to ctc downstream of prs. cDNA synthesis with a B. caldolyticus gcaD-prs-ctc-specified mRNA as template, followed by amplification utilising the polymerase chain reaction indicated that the three genes are co......-transcribed. Comparison of amino acid sequences revealed a high similarity among PRPP synthases across a wide phylogenetic range. An E. coli strain harbouring the B. caldolyticus prs gene in a multicopy plasmid produced PRPP synthase activity 33-fold over the activity of a haploid B. caldolyticus strain. B. caldolyticus...

  15. Stereoselective Copolymerization of Styrene with Terpenes Catalyzed by an Ansa-Lanthanidocene Catalyst: Access to New Syndiotactic Polystyrene-Based Materials

    Directory of Open Access Journals (Sweden)

    Eva Laur

    2017-11-01

    Full Text Available The copolymerization of bio-renewable β-myrcene or β-farnesene with styrene was examined using an ansa-neodymocene catalyst, affording two series of copolymers with high styrene content and unprecedented syndioregularity of the polystyrene sequences. The incorporation of terpene in the copolymers ranged from 5.6 to 30.8 mol % (β-myrcene and from 2.5 to 9.8 mol % (β-farnesene, respectively. NMR spectroscopy and DSC analyses suggested that the microstructure of the copolymers consists of 1,4- and 3,4-poly(terpene units randomly distributed along syndiotactic polystyrene chains. The thermal properties of the copolymers are strongly dependent on the terpene content, which is easily controlled by the initial feed. The terpolymerization of styrene with β-myrcene in the presence of ethylene was also examined.

  16. The FEI-TPS on the Upper Surface of the X-38

    Science.gov (United States)

    Antonenko, Johann; Kowal, John

    2002-01-01

    The X-38 is being developed by NASA-JSC as a technology demonstrator of a future Crew Rescue Vehicle. The size of the vehicle is limited to fit into the cargo bay of the shuttle. Due to its small size and shuttle-like trajectory all surfaces will receive comparably high heat rates leading to high surface temperatures. Temperatures on the nose are calculated to reach 1750°C, which is significantly higher than on the shuttle. Due to the lifting body shape, large areas of the central fuselage will be exposed to flow of hot gases around the vehicle. Here temperatures of the upper surface are calculated to reach up to 1000°C and the application of a high temperature blanket thermal protection system (TPS) becomes mandatory. Consecutively, the temperature level of the upper surface and the base area will be significantly high. Unlike on the shuttle, where large areas of the surface are covered by flexible reusable surface insulation (FRSI), locations with temperatures below 400°C will be scarce on the X-38. During development of the European shuttle HERMES the Flexible External Insulation (FEI) was developed for the upper surface TPS. This development was continued by ESA and DLR funded programs and currently a product family is available for temperatures ranging from 450°C to 1100°C for re-usable application. For a single re- entry under ultimate conditions temperatures may reach up to 1400°C. Under funding of DLR and ESA, the FEI assembly is one of the European contributions to the X-38. Three subassemblies have been chosen: the FEI-450, FEI-650 and FEI- 1000, capable of limit temperatures of 450°C, 650°C and 1000°C, respectively. The FEI-650 and FEI-1000 were already developed in the HERMES program. The FEI- 450 was developed in the German TETRA program. The qualification for X-38 application was performed for temperatures up to 510°C for the FEI-450 and up to 1130°C for the FEI-1000. Acoustic noise loads of up to 160dB have been endured, far beyond what X

  17. The Transcriptome and Terpene Profile of Eucalyptus grandis Reveals Mechanisms of Defense Against the Insect Pest, Leptocybe invasa.

    Science.gov (United States)

    Oates, Caryn N; Külheim, Carsten; Myburg, Alexander A; Slippers, Bernard; Naidoo, Sanushka

    2015-07-01

    Plants have evolved complex defenses that allow them to protect themselves against pests and pathogens. However, there is relatively little information regarding the Eucalyptus defensome. Leptocybe invasa is one of the most damaging pests in global Eucalyptus forestry, and essentially nothing is known regarding the molecular mechanisms governing the interaction between the pest and host. The aim of the study was to investigate changes in the transcriptional landscape and terpene profile of a resistant and susceptible Eucalyptus genotype in an effort to improve our understanding of this interaction. We used RNA-seqencing to investigate transcriptional changes following L. invasa oviposition. Expression levels were validated using real-time quantitative PCR. Terpene profiles were investigated using gas chromatography coupled to mass spectometry on uninfested and oviposited leaves. We found 698 and 1,115 significantly differentially expressed genes from the resistant and susceptible interactions, respectively. Gene Ontology enrichment and Mapman analyses identified putative defense mechanisms including cell wall reinforcement, protease inhibitors, cell cycle suppression and regulatory hormone signaling pathways. There were significant differences in the mono- and sesquiterpene profiles between genotypes and between control and infested material. A model of the interaction between Eucalyptus and L. invasa was proposed from the transcriptomic and chemical data. © 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.

  18. Morphological analysis of thermoplastic starch films and montmorillonite (TPS/MMT) using vegetable oils of Brazilian Cerrado as plasticizers;Morfologia de filmes de amido termoplastico e montmorilonita (TPS/MMT) usando oleos vegetais do cerrado como plastificantes

    Energy Technology Data Exchange (ETDEWEB)

    Schlemmer, Daniela; Sales, Maria Jose A. [Universidade de Brasilia (UnB), DF (Brazil). Lab. de Pesquisa em Polimeros; Angelica, Romulo S. [Universidade Federal do Para (UFPA), Belem, PA (Brazil). Lab. de Raios X; Gomes, Ana Cristina M.M. [Empresa Brasileira de Pesquisa Agropecuaria (EMBRAPA-CENARGEN), Brasilia, DF (Brazil). Lab. de Microscopia Eletronica

    2009-07-01

    Biopolymers can be used where petrochemical plastics have applications with short life. The excellent degradation of starch and its low cost make it an alternative for obtaining biodegradable plastics. To obtain thermoplastic starch (TPS) is necessary mechanical shake, high temperature and plasticizers. In this work, TPS were produced using three different vegetable oils from Brazilian's cerrado as plasticizers: buriti, macauba or pequi. Materials are also produced with montmorillonite (MMT). All the materials were analyzed by scanning electronic microscopy (SEM) and X-ray diffraction (XRD). The starch micrographs revealed irregular delaminate with a predominance of starch and 'holes' for the oils. In nanocomposites the clusters of clay are dispersed without a defined standard. It was understood that the plasticizers and processing completely changed the structure of starch causing a decrease in their crystallinity degree. Almost all nanocomposites presented exfoliate structure, only one presented intercalated structure. (author)

  19. Cloning and expression of pineapple sucrosephosphate synthase ...

    African Journals Online (AJOL)

    A 1132-base pairs (bp) polymerase-chain-reaction product of sucrose-phosphate synthase (SPS) (EC 2.3.1.14) from pineapple (Ananas comosus cv. Comte de paris) fruit was cloned and nominated as Ac- SPS1. The sequence encodes a putative 377 amino acids protein containing two serine conserved features that had ...

  20. Endothelial nitric oxide synthase gene polymorphisms associated ...

    African Journals Online (AJOL)

    Endothelial nitric oxide synthase (NOS3) is involved in key steps of immune response. Genetic factors predispose individuals to periodontal disease. This study's aim was to explore the association between NOS3 gene polymorphisms and clinical parameters in patients with periodontal disease. Genomic DNA was obtained ...

  1. HAEM SYNTHASE AND COBALT PORPHYRIN SYNTHASE IN VARIOUS MICRO-ORGANISMS.

    Science.gov (United States)

    PORRA, R J; ROSS, B D

    1965-03-01

    1. The preparation of a crude extract of Clostridium tetanomorphum containing cobalt porphyrin synthase but little haem-synthase activity is described. 2. The properties of cobalt porphyrin synthase in the clostridial extracts is compared with the properties of a haem synthase present in crude extracts of the yeast Torulopsis utilis. 3. Cobalt porphyrin synthase in extracts of C. tetanomorphum inserts Co(2+) ions into the following dicarboxylic porphyrins in descending order of rate of insertion: meso-, deutero- and proto-porphyrins. Esterification renders meso- and deutero-porphyrins inactive as substrates. Neither the tetracarboxylic (coproporphyrin III) nor the octacarboxylic (uroporphyrin III) compounds are converted into cobalt porphyrins by the extract, but the non-enzymic incorporation of Co(2+) ions into these two porphyrins is rapid. These extracts are unable to insert Mn(2+), Zn(2+), Mg(2+) or Cu(2+) ions into mesoporphyrin. 4. Crude extracts of T. utilis readily insert both Co(2+) and Fe(2+) ions into deutero-, meso, and proto-porphyrins. Unlike the extracts of C. tetanomorphum, these preparations catalyse the insertion of Co(2+) ions into deuteroporphyrin more rapidly than into mesoporphyrin. This parallels the formation of haems by the T. utilis extract. 5. Cobalt porphyrin synthase is present in the particulate fraction of the extracts of C. tetanomorphum but requires a heat-stable factor present in the soluble fraction. This soluble factor can be replaced by GSH. 6. Cobalt porphyrin synthase in the clostridial extract is inhibited by iodoacetamide and to a smaller extent by p-chloromercuribenzoate and N-ethylmaleimide. The haem synthases of T. utilis and Micrococcus denitrificans are also inhibited by various thiol reagents.

  2. Independent dose calculation in IMRT for the Tps Iplan using the Clarkson modified integral; Calculo independiente de dosis en IMRT para el TPS Iplan usando la integral modificada de Clarkson

    Energy Technology Data Exchange (ETDEWEB)

    Adrada, A.; Tello, Z.; Garrigo, E.; Venencia, D., E-mail: jorge.alberto.adrada@gmail.com [Instituto Privado de Radioterapia, Obispo Oro 423, X5000BFI Cordoba (Argentina)

    2014-08-15

    Intensity-Modulated Radiation Therapy (IMRT) treatments require a quality assurance (Q A) specific patient before delivery. These controls include the experimental verification in dose phantom of the total plan as well as dose distributions. The use of independent dose calculation (IDC) is used in 3D-Crt treatments; however its application in IMRT requires the implementation of an algorithm that allows considering a non-uniform intensity beam. The purpose of this work was to develop IDC software in IMRT with MLC using the algorithm proposed by Kung (Kung et al. 2000). The software was done using Matlab programming. The Clarkson modified integral was implemented on each flowing, applying concentric rings for the dose determination. From the integral of each field was calculated the dose anywhere. One time finished a planning; all data are exported to a phantom where a Q A plan is generated. On this is calculated the half dose in a representative volume of the ionization chamber and the dose at the center of it. Until now 230 IMRT planning were analyzed carried out ??in the treatment planning system (Tps) Iplan. For each one of them Q A plan was generated, were calculated and compared calculated dose with the Tps, IDC system and measurement with ionization chamber. The average difference between measured and calculated dose with the IDC system was 0.4% ± 2.2% [-6.8%, 6.4%]. The difference between the measured and the calculated doses by the pencil-beam algorithm (Pb) of Tps was 2.6% ± 1.41% [-2.0%, 5.6%] and with the Monte Carlo algorithm was 0.4% ± 1.5% [-4.9%, 3.7%]. The differences of the carried out software are comparable to the obtained with the ionization chamber and Tps in Monte Carlo mode. (author)

  3. Morphological analysis of thermoplastic starch films and montmorillonite (TPS/MMT) using vegetable oils of Brazilian Cerrado as plasticizers

    International Nuclear Information System (INIS)

    Schlemmer, Daniela; Sales, Maria Jose A.; Angelica, Romulo S.; Gomes, Ana Cristina M.M.

    2009-01-01

    Biopolymers can be used where petrochemical plastics have applications with short life. The excellent degradation of starch and its low cost make it an alternative for obtaining biodegradable plastics. To obtain thermoplastic starch (TPS) is necessary mechanical shake, high temperature and plasticizers. In this work, TPS were produced using three different vegetable oils from Brazilian's cerrado as plasticizers: buriti, macauba or pequi. Materials are also produced with montmorillonite (MMT). All the materials were analyzed by scanning electronic microscopy (SEM) and X-ray diffraction (XRD). The starch micrographs revealed irregular delaminate with a predominance of starch and 'holes' for the oils. In nanocomposites the clusters of clay are dispersed without a defined standard. It was understood that the plasticizers and processing completely changed the structure of starch causing a decrease in their crystallinity degree. Almost all nanocomposites presented exfoliate structure, only one presented intercalated structure. (author)

  4. EFEKTIVITAS PENERAPAN MODEL PEMBELAJARAN THINK PAIR SHARE (TPS TERHADAP HASIL BELAJAR DAN AKTIVITAS SISWA PADA MATERI SISTEM PERNAPASAN MANUSIA

    Directory of Open Access Journals (Sweden)

    Maria Yashinta Afoan

    2016-10-01

    Tujuan dari penelitian ini adalah untuk mengetahui efektivitas penerapan model pembelajaran kooperatif melalui pendekatan Think Pair Share terhadap hasil belajar dan aktivitas siswa. Data dikumpulkan melalui wawancara dan tes esai hasil belajar. Data hasil wawancara dianalisis secara deskriptif kualitatif, sedangkan tes esai hasil belajar dianalisis secara deskriptif kuantitatif. Hasil penelitian menunjukkan bahwa penerapan model pembelajaran Think Pair Share (TPS efektif dapat meningkatkan hasil belajar siswa. Hal ini dilihat dari rata-rata pretest sebesar 34,06% mengalami peningkatan pada posttest sebesar 83,13 % dengan rata-rata peningkatann pretest ke posttest sebesar 49,06 %, dan ketuntasan klasikal hasil belajar sebesar 87,50% begitu juga dengan penerapan model pembelajaran Think Pair Share (TPS aktivitas belajar siswa dapat ditingkatkan.

  5. Monte Carlo characterization of materials for prosthetic implants and dosimetric validation of Pinnacle3 TPS

    International Nuclear Information System (INIS)

    Palleri, Francesca; Baruffaldi, Fabio; Angelini, Anna Lisa; Ferri, Andrea; Spezi, Emiliano

    2008-01-01

    In external beam radiotherapy the calculation of dose distribution for patients with hip prostheses is critical. Metallic implants not only degrade the image quality but also perturb the dose distribution. Conventional treatment planning systems do not accurately account for high-Z prosthetic implants heterogeneities, especially at interfaces. The materials studied in this work have been chosen on the basis of a statistical investigation on the hip prostheses implanted in 70 medical centres. The first aim of this study is a systematic characterization of materials used for hip prostheses, and it has been provided by BEAMnrc Monte Carlo code. The second aim is to evaluate the capabilities of a specific treatment planning system, Pinnacle 3 , when dealing with dose calculations in presence of metals, also close to the regions of high-Z gradients. In both cases it has been carried out an accurate comparison versus experimental measurements for two clinical photon beam energies (6 MV and 18 MV) and for two experimental sets-up: metallic cylinders inserted in a water phantom and in a specifically built PMMA slab. Our results show an agreement within 2% between experiments and MC simulations. TPS calculations agree with experiments within 3%.

  6. Monte Carlo characterization of materials for prosthetic implants and dosimetric validation of Pinnacle 3 TPS

    Science.gov (United States)

    Palleri, Francesca; Baruffaldi, Fabio; Angelini, Anna Lisa; Ferri, Andrea; Spezi, Emiliano

    2008-12-01

    In external beam radiotherapy the calculation of dose distribution for patients with hip prostheses is critical. Metallic implants not only degrade the image quality but also perturb the dose distribution. Conventional treatment planning systems do not accurately account for high-Z prosthetic implants heterogeneities, especially at interfaces. The materials studied in this work have been chosen on the basis of a statistical investigation on the hip prostheses implanted in 70 medical centres. The first aim of this study is a systematic characterization of materials used for hip prostheses, and it has been provided by BEAMnrc Monte Carlo code. The second aim is to evaluate the capabilities of a specific treatment planning system, Pinnacle 3, when dealing with dose calculations in presence of metals, also close to the regions of high-Z gradients. In both cases it has been carried out an accurate comparison versus experimental measurements for two clinical photon beam energies (6 MV and 18 MV) and for two experimental sets-up: metallic cylinders inserted in a water phantom and in a specifically built PMMA slab. Our results show an agreement within 2% between experiments and MC simulations. TPS calculations agree with experiments within 3%.

  7. Design and fabrication of the vacuum systems for TPS pulsed septum magnets

    Energy Technology Data Exchange (ETDEWEB)

    Chan, C.K.; Chang, C.C.; Chen, C.L.; Yang, C.S.; Chen, C.S.; Lin, F.Y.; Chen, J.R.

    2014-11-01

    Three in-air pulsed septum magnets were developed to inject and extract electron beams for the 3 GeV synchrotron facility, the Taiwan Photon Source (TPS). The vacuum chamber is a novel combined aluminium-stainless steel design, using a bimetallic flange to connect the two material types. To evaluate the vacuum performances of these vacuum chambers, we set up a test bench at which we simultaneously measure the rates of thermal outgassing of the aluminium chamber and the septum tube with a throughput method. The test result indicates that the rate q{sub 72} of thermal outgassing measured after 1 day from baking at 150 °C was 1×10{sup −13} mbar L s{sup −1} cm{sup −2}. The magnetic leakage measurements show the combination of conductor slitting, magnetic shielding and the aluminium vacuum chamber reduce the peak value of the leakage field integral to ∼10 G cm along the trajectory of the stored beam.

  8. Development and validation of a kit for radioimmunometric determination of TPS

    International Nuclear Information System (INIS)

    Berrueta Robles, A.M.

    1997-01-01

    In recent years several tumour markers have been investigated to assess their value as a tool for early detection or follow up after or during treatment. It is important to estimate the role of analytical procedures on the results of biological markers. IRMA has been recognized as a better and more sensitive system than RIA. Also non isotopic techniques compared with radioactive methodologies are considered less sensitives. Early detection is closely related with the reliability and sensitivity of the methodologies used. Among analytical procedures in current use IRMA with monoclonal antibodies immobilized to a solid matrix (coated tube) are first in specificity and sensitivity. Detailed objectives of research: Develop a kit for the determination of TPS and assess the relevance as tumor marker in breast cancer; Study the parameters related with design of the analytical system and its components; Evaluate and select control methods for each development step; Perform the assay on patients clinically diagnosed with the pathology according with a protocol designed with oncologists

  9. Penerapan Model Pembelajaran TPS Untuk Meningkatkan Hasil Belajar IPA Siswa Kelas V SD No 1 Mengwitani

    Directory of Open Access Journals (Sweden)

    I W Daniel Winantara

    2017-05-01

    Full Text Available Tujuan penelitian ini untuk mengetahui peningkatan hasil belajar IPA siswa kelas V SD No. 1 Mengwitani setelah diterapkannya model pembelajaran think pair share (TPS. Penelitian ini adalah penelitian tindakan kelas yang dilakukan dalam dua siklus. Subjek dalam penelitian ini adalah siswa kelas V yang berjumlah 32 orang. Data penelitian ini tentang hasil belajar yang dikumpulkan dengan menggunakan metode tes. Data dianalisis dengan menggunakan statistik kuantitatif. Hasil penelitian menunjukkan adanya peningkatan terhadap hasil belajar IPA siswa kelas V SD No. 1 Mengwitani. Hal ini ditunjukkan dengan hasil penelitian yang diperoleh yaitu persentase rata-rata hasil belajar pada siklus I sebesar 75,31% yang berada pada kategori sedang dan pada siklus II menjadi 80,15% yang berada pada kategori tinggi. Dengan ketuntasan belajar siswa pada siklus I sebesar 65,62% dan pada siklus II mencapai 87,5%. Berdasarkan hasil penelitian tersebut dapat disimpulkan penerapan model pembelajaran think pair share dapat meningkatkan hasil belajar IPA siswa kelas V SD No. 1 Mengwitani.

  10. Molecular cloning and characterization of a cDNA encoding the gibberellin biosynthetic enzyme ent-kaurene synthase B from pumpkin (Cucurbita maxima L.).

    Science.gov (United States)

    Yamaguchi, S; Saito, T; Abe, H; Yamane, H; Murofushi, N; Kamiya, Y

    1996-08-01

    The first committed step in the formation of diterpenoids leading to gibberellin (GA) biosynthesis is the conversion of geranylgeranyl diphosphate (GGDP) to ent-kaurene. ent-Kaurene synthase A (KSA) catalyzes the conversion of GGDP to copalyl diphosphate (CDP), which is subsequently converted to ent-kaurene by ent-kaurene synthase B (KSB). A full-length KSB cDNA was isolated from developing cotyledons in immature seeds of pumpkin (Cucurbita maxima L.). Degenerate oligonucleotide primers were designed from the amino acid sequences obtained from the purified protein to amplify a cDNA fragment, which was used for library screening. The isolated full-length cDNA was expressed in Escherichia coli as a fusion protein, which demonstrated the KSB activity to cyclize [3H]CDP to [3H]ent-kaurene. The KSB transcript was most abundant in growing tissues, but was detected in every organ in pumpkin seedlings. The deduced amino acid sequence shares significant homology with other terpene cyclases, including the conserved DDXXD motif, a putative divalent metal ion-diphosphate complex binding site. A putative transit peptide sequence that may target the translated product into the plastids is present in the N-terminal region.

  11. SU-E-T-169: Evaluation of Oncentra TPS for Nasopharynx Brachy Using Patient Specific Voxel Phantom and EGSnrc

    Energy Technology Data Exchange (ETDEWEB)

    Hadad, K; Zoherhvand, M; Faghihi, R [Shiraz University, Shiraz (Iran, Islamic Republic of)

    2014-06-01

    Purpose: Nasopharnx carcinoma (NPC) treatment is being carried out using Ir-192 HDR seeds in Mehdieh Hospital in Hamadan, Iran. The Oncentra™ TPS is based on optimized TG-43 formalism which disregards heterogeneity in the treatment area. Due to abundant heterogeneity in head and neck, comparison of the Oncentra™ TPS dose evaluation and an accurate dose calculation method in NPC brachytherapy is the objective of this study. Methods: CT DICOMs of a patient with NPC obtained from Mehdieh Hospital used to create 3D voxel phantom with CTCREATE utility of EGSnrc code package. The voxel phantom together with Ir-192 HDR brachytherapy source were the input to DOSXYZnrc to calculate the 3D dose distribution. The sources were incorporate with type 6 source in DOSXYZnrc and their dwell times were taken into account in final dose calculations. Results: The direct comparison between isodoses as well as DVHs for the GTV, PTV and CTV obtained by Oncentra™ and EGSnrc Monte Carlo code are made. EGSnrc results are obtained using 5×10{sup 9} histories to reduce the statistical error below 1% in GTV and 5% in 5% dose areas. The standard ICRP700 cross section library is employed in DOSXYZnrc dose calculation. Conclusion: A direct relationship between increased dose differences and increased material density (hence heterogeneity) is observed when isodoses contours of the TPS and DOSXYZnrc are compared. Regarding the point dose calculations, the differences range from 1.2% in PTV to 5.6% for cavity region and 7.8% for bone regions. While Oncentra™ TPS overestimates the dose in cavities, it tends to underestimate dose depositions within bones.

  12. SU-E-T-169: Evaluation of Oncentra TPS for Nasopharynx Brachy Using Patient Specific Voxel Phantom and EGSnrc

    International Nuclear Information System (INIS)

    Hadad, K; Zoherhvand, M; Faghihi, R

    2014-01-01

    Purpose: Nasopharnx carcinoma (NPC) treatment is being carried out using Ir-192 HDR seeds in Mehdieh Hospital in Hamadan, Iran. The Oncentra™ TPS is based on optimized TG-43 formalism which disregards heterogeneity in the treatment area. Due to abundant heterogeneity in head and neck, comparison of the Oncentra™ TPS dose evaluation and an accurate dose calculation method in NPC brachytherapy is the objective of this study. Methods: CT DICOMs of a patient with NPC obtained from Mehdieh Hospital used to create 3D voxel phantom with CTCREATE utility of EGSnrc code package. The voxel phantom together with Ir-192 HDR brachytherapy source were the input to DOSXYZnrc to calculate the 3D dose distribution. The sources were incorporate with type 6 source in DOSXYZnrc and their dwell times were taken into account in final dose calculations. Results: The direct comparison between isodoses as well as DVHs for the GTV, PTV and CTV obtained by Oncentra™ and EGSnrc Monte Carlo code are made. EGSnrc results are obtained using 5×10 9 histories to reduce the statistical error below 1% in GTV and 5% in 5% dose areas. The standard ICRP700 cross section library is employed in DOSXYZnrc dose calculation. Conclusion: A direct relationship between increased dose differences and increased material density (hence heterogeneity) is observed when isodoses contours of the TPS and DOSXYZnrc are compared. Regarding the point dose calculations, the differences range from 1.2% in PTV to 5.6% for cavity region and 7.8% for bone regions. While Oncentra™ TPS overestimates the dose in cavities, it tends to underestimate dose depositions within bones

  13. Recommendations for the commissioning and use of a radiotherapy treatment planning system (TPS). S.F.P.M. report nr 27, December 1, 2010

    International Nuclear Information System (INIS)

    Rosenwald, Jean Claude; Bonvalet, Laurent; Mazurier, Jocelyne; Metayer, Christine; Beaudre, Anne; Garcia, Robin; Ruchaud, Romain; Dedieu, Veronique; Bramoulle, Celine; Caselles, Olivier; Lacaze, Brigitte; Mazurier, Jocelyne

    2010-01-01

    This report aims at bringing risks related to the use of Treatment Planning Systems (TPS) to the attention of French medical physicists, and to provide them with practical, precise and updated recommendations related to new legal requirements on commissioning and use of a new planning system. It addresses TPS used in external radiation therapy. It is based on various international (IAEA, ESTRO, AAPM, and so on) and national recommendations, and proposes some rules of good practices for the implementation of a quality insurance policy as well as for the procurement and implementation of a new planning system, as for the performance follow-up of an already installed system. The authors address the following issues: risks associated with the use of a TPS, organisational aspects, dose calculation algorithms and required precision, TPS acceptance, adjustment and tests, periodic controls, treatment plan verification procedures

  14. Transient plane source (tps) sensors for simultaneous measurements of thermal conductivity and thermal diffusivity of insulators, fluids and conductors

    Science.gov (United States)

    Maqsood, Asghari; Anis-ur-Rehman, M.

    2013-12-01

    Thermal conductivity and thermal diffusivity are two important physical properties for designing any food engineering processes1. The knowledge of thermal properties of the elements, compounds and different materials in many industrial applications is a requirement for their final functionality. Transient plane source (tps) sensors are reported2 to be useful for the simultaneous measurement of thermal conductivity, thermal diffusivity and volumetric heat capacity of insulators, conductor liquids3 and high-TC superconductors4. The tps-sensor consists of a resistive element in the shape of double spiral made of 10 micrometer thick Ni-foils covered on both sides with 25 micrometer thick Kapton. This sensor acts both as a heat source and a resistance thermometer for recording the time dependent temperature increase. From the knowledge of the temperature co-efficient of the metal spiral, the temperature increase of the sensor can be determined precisely by placing the sensor in between two surfaces of the same material under test. This temperature increase is then related to the thermal conductivity, thermal diffusivity and volumetric heat capacity by simple relations2,5. The tps-sensor has been used to measure thermal conductivities from 0.001 Wm-1K-1to 600 Wm-1K-1 and temperature ranges covered from 77K- 1000K. This talk gives the design, advantages and limitations of the tpl-sensor along with its applications to the measurementof thermal properties in a variety of materials.

  15. Transient plane source (tps) sensors for simultaneous measurements of thermal conductivity and thermal diffusivity of insulators, fluids and conductors

    International Nuclear Information System (INIS)

    Maqsood, Asghari; Anis-ur-Rehman, M

    2013-01-01

    Thermal conductivity and thermal diffusivity are two important physical properties for designing any food engineering processes 1 . The knowledge of thermal properties of the elements, compounds and different materials in many industrial applications is a requirement for their final functionality. Transient plane source (tps) sensors are reported 2 to be useful for the simultaneous measurement of thermal conductivity, thermal diffusivity and volumetric heat capacity of insulators, conductor liquids 3 and high-T C superconductors 4 . The tps-sensor consists of a resistive element in the shape of double spiral made of 10 micrometer thick Ni-foils covered on both sides with 25 micrometer thick Kapton. This sensor acts both as a heat source and a resistance thermometer for recording the time dependent temperature increase. From the knowledge of the temperature co-efficient of the metal spiral, the temperature increase of the sensor can be determined precisely by placing the sensor in between two surfaces of the same material under test. This temperature increase is then related to the thermal conductivity, thermal diffusivity and volumetric heat capacity by simple relations 2,5 . The tps-sensor has been used to measure thermal conductivities from 0.001 Wm −1 K −1 to 600 Wm −1 K −1 and temperature ranges covered from 77K– 1000K. This talk gives the design, advantages and limitations of the tpl-sensor along with its applications to the measurementof thermal properties in a variety of materials

  16. Long-Term Persistence of Pesticides and TPs in Archived Agricultural Soil Samples and Comparison with Pesticide Application.

    Science.gov (United States)

    Chiaia-Hernandez, Aurea C; Keller, Armin; Wächter, Daniel; Steinlin, Christine; Camenzuli, Louise; Hollender, Juliane; Krauss, Martin

    2017-09-19

    For polar and more degradable pesticides, not many data on long-term persistence in soil under field conditions and real application practices exist. To assess the persistence of pesticides in soil, a multiple-compound screening method (log K ow 1.7-5.5) was developed based on pressurized liquid extraction, QuEChERS and LC-HRMS. The method was applied to study 80 polar pesticides and >90 transformation products (TPs) in archived topsoil samples from the Swiss Soil Monitoring Network (NABO) from 1995 to 2008 with known pesticide application patterns. The results reveal large variations between crop type and field sites. For the majority of the sites 10-15 pesticides were identified with a detection rate of 45% at concentrations between 1 and 330 μg/kg dw in soil. Furthermore, TPs were detected in 47% of the cases where the "parent-compound" was applied. Overall, residues of about 80% of all applied pesticides could be detected with half of these found as TPs with a persistence of more than a decade.

  17. The effect of Think Pair Share (TPS) using scientific approach on students’ self-confidence and mathematical problem-solving

    Science.gov (United States)

    Rifa’i, A.; Lestari, H. P.

    2018-03-01

    This study was designed to know the effects of Think Pair Share using Scientific Approach on students' self-confidence and mathematical problem-solving. Quasi-experimental with pre-test post-test non-equivalent group method was used as a basis for design this study. Self-confidence questionnaire and problem-solving test have been used for measurement of the two variables. Two classes of the first grade in religious senior high school (MAN) in Indonesia were randomly selected for this study. Teaching sequence and series from mathematics book at control group in the traditional way and at experiment group has been in TPS using scientific approach learning method. For data analysis regarding students’ problem-solving skill and self-confidence, One-Sample t-Test, Independent Sample t-Test, and Multivariate of Variance (MANOVA) were used. The results showed that (1) TPS using a scientific approach and traditional learning had positive effects (2) TPS using scientific approach learning in comparative with traditional learning had a more significant effect on students’ self-confidence and problem-solving skill.

  18. EFEKTIVITAS PEMBELAJARAN MATEMATIKA MENGGUNAKAN METODE TPS (THINK-PAIR-SHARE BERBASIS OPEN-ENDED-PROBLEM TERHADAP KREATIVITAS BELAJAR SISWA

    Directory of Open Access Journals (Sweden)

    Sri Hudia Rohmatin

    2015-08-01

    Full Text Available This study was aimed at finding out whether or not Open-Ended-Problem Based TPS Method was effective on student’s mathematics learning creativity. The type of this research was quasi-eksperimental with one-group pretet posttest desaign. The sample of this research was 33 students. Instrument in form of 4 essay that was valid and reliable. Normality testing with Chi-kuadrat and homogeneity testing with Barlett-test  while hypotesis testing with paramteric statistical analysis t-test. Based on the data analysis average pre test was 44 and post test was 64. The results of the normality testing pre-test group obtained x2count < x2tabel or 4.452  < 11.070 and post test group obtained x2count < x2tabel or 5,946 < 11.070. Data homogeneity testing results obtained x2count < x2tabel 2,484  < 3,841. The results of hypothesis testing data was obtained tcount > ttabel or 12,317 > 2,001. Based on the test results concluded that the hypothesis Ho was rejected and Ha accepted. To sum up, Open-Ended-Problem-Based TPS was significantly effective on student’s mathematics learning creativity. Key words: effectiveness, TPS method, open-ended-problem, students creativity DOI: http://dx.doi.org/10.22342/jpm.9.2.2085.178 - 187

  19. Development of virtual CT DICOM images of patients with tumors: application for TPS and Monte Carlo dose evaluation

    International Nuclear Information System (INIS)

    Milian, F. M.; Attili, A.; Russo, G; Marchetto, F.; Cirio, R.; Bourhaleb, F.

    2013-01-01

    A novel procedure for the generation of a realistic virtual Computed Tomography (CT) image of a patient, using the advanced Boundary RE Presentation (BREP)-based model MASH, has been implemented. This method can be used in radiotherapy assessment. It is shown that it is possible to introduce an artificial cancer, which can be modeled using mesh surfaces. The use of virtual CT images based on BREP models presents several advantages with respect to CT images of actual patients, such as automation, control and flexibility. As an example, two artificial cases, namely a brain and a prostate cancer, were created through the generation of images and tumor/organ contours. As a secondary objective, the described methodology has been used to generate input files for treatment planning system (TPS) and Monte Carlo code dose evaluation. In this paper, we consider treatment plans generated assuming a dose delivery via an active proton beam scanning performed with the INFN-IBA TPS kernel. Additionally, Monte Carlo simulations of the two treatment plans were carried out with GATE/GEANT4. The work demonstrates the feasibility of the approach based on the BREP modeling to produce virtual CT images. In conclusion, this study highlights the benefits in using digital phantom model capable of representing different anatomical structures and varying tumors across different patients. These models could be useful for assessing radiotherapy treatment planning systems (TPS) and computer simulations for the evaluation of the adsorbed dose. (author)

  20. Preparation of sulfonated graphene/polypyrrole solid-phase microextraction coating by in situ electrochemical polymerization for analysis of trace terpenes.

    Science.gov (United States)

    Zhang, Chengjiang; Zhang, Zhuomin; Li, Gongke

    2014-06-13

    In this study, a novel sulfonated graphene/polypyrrole (SG/PPy) solid-phase microextraction (SPME) coating was prepared and fabricated on a stainless-steel wire by a one-step in situ electrochemical polymerization method. Crucial preparation conditions were optimized as polymerization time of 15min and SG doping amount of 1.5mg/mL. SG/PPy coating showed excellent thermal stability and mechanical durability with a long lifespan of more than 200 stable replicate extractions. SG/PPy coating demonstrated higher extraction selectivity and capacity to volatile terpenes than commonly-used commercial coatings. Finally, SG/PPy coating was practically applied for the analysis of volatile components from star anise and fennel samples. The majority of volatile components identified were terpenes, which suggested the ultra-high extraction selectivity of SG/PPy coating to terpenes during real analytical projects. Four typical volatile terpenes were further quantified to be 0.2-27.4μg/g from star anise samples with good recoveries of 76.4-97.8% and 0.1-1.6μg/g from fennel samples with good recoveries of 80.0-93.1%, respectively. Copyright © 2014 Elsevier B.V. All rights reserved.

  1. Mountain pine beetles colonizing historical and naive host trees are associated with a bacterial community highly enriched in genes contributing to terpene metabolism.

    Science.gov (United States)

    Adams, Aaron S; Aylward, Frank O; Adams, Sandye M; Erbilgin, Nadir; Aukema, Brian H; Currie, Cameron R; Suen, Garret; Raffa, Kenneth F

    2013-06-01

    The mountain pine beetle, Dendroctonus ponderosae, is a subcortical herbivore native to western North America that can kill healthy conifers by overcoming host tree defenses, which consist largely of high terpene concentrations. The mechanisms by which these beetles contend with toxic compounds are not well understood. Here, we explore a component of the hypothesis that beetle-associated bacterial symbionts contribute to the ability of D. ponderosae to overcome tree defenses by assisting with terpene detoxification. Such symbionts may facilitate host tree transitions during range expansions currently being driven by climate change. For example, this insect has recently breached the historical geophysical barrier of the Canadian Rocky Mountains, providing access to näive tree hosts and unprecedented connectivity to eastern forests. We use culture-independent techniques to describe the bacterial community associated with D. ponderosae beetles and their galleries from their historical host, Pinus contorta, and their more recent host, hybrid P. contorta-Pinus banksiana. We show that these communities are enriched with genes involved in terpene degradation compared with other plant biomass-processing microbial communities. These pine beetle microbial communities are dominated by members of the genera Pseudomonas, Rahnella, Serratia, and Burkholderia, and the majority of genes involved in terpene degradation belong to these genera. Our work provides the first metagenome of bacterial communities associated with a bark beetle and is consistent with a potential microbial contribution to detoxification of tree defenses needed to survive the subcortical environment.

  2. Eksprimentasi Pembelajaran Kooperatif Tipe Stad dan Tps dengan Pendekatan Ctl pada Materi Pokok Sistem Persamaan Linear Dua Variabel Ditinjau dari Gaya Belajar Siswa

    OpenAIRE

    Zamroni, Zamroni; Budiyono, Budiyono; Sujadi, Imam

    2013-01-01

    The objective of research was to find out: (1) which ones having better achievement, the students using TPS (Think Pair Share) learning with CTL approach or STAD (Students Teams Achievement Divisions) cooperative learning model with CTL approach or direct learning, (2) which ones having better learning achievement, the students with kinesthetic or visual or auditory learning style, (3) in each learning style, which ones having better achievement, the students using TPS (Think Pair Share) lear...

  3. A Possible Trifunctional β-Carotene Synthase Gene Identified in the Draft Genome of Aurantiochytrium sp. Strain KH105

    Directory of Open Access Journals (Sweden)

    Hiroaki Iwasaka

    2018-04-01

    Full Text Available Labyrinthulomycetes have been regarded as a promising industrial source of xanthophylls, including astaxanthin and canthaxanthin, polyunsaturated fatty acids such as docosahexaenoic acid and docosapentaenoic acid, ω-3 oils, and terpenic hydrocarbons, such as sterols and squalene. A Thraustochytrid, Aurantiochytrium sp. KH105 produces carotenoids, including astaxanthin, with strong antioxidant activity. To gain genomic insights into this capacity, we decoded its 97-Mbp genome and characterized genes for enzymes involved in carotenoid biosynthesis. Interestingly, all carotenogenic genes, as well as other eukaryotic genes, appeared duplicated, suggesting that this strain is diploid. In addition, among the five genes involved in the pathway from geranylgeranyl pyrophosphate to astaxanthin, geranylgeranyl phytoene synthase (crtB, phytoene desaturase (crtI and lycopene cyclase (crtY were fused into single gene (crtIBY with no internal stop codons. Functionality of the trifunctional enzyme, CrtIBY, to catalyze the reaction from geranylgeranyl diphosphate to β-carotene was confirmed using a yeast assay system and mass spectrometry. Furthermore, analyses of differential gene expression showed characteristic up-regulation of carotenoid biosynthetic genes during stationary and starvation phases under these culture conditions. This suggests genetic engineering events to promote more efficient production of carotenoids. We also showed an occurrence of crtIBY in other Thraustochytrid species.

  4. Glycogen Phosphorylase and Glycogen Synthase: Gene Cloning and Expression Analysis Reveal Their Role in Trehalose Metabolism in the Brown Planthopper, Nilaparvata lugens Stål (Hemiptera: Delphacidae).

    Science.gov (United States)

    Zhang, Lu; Wang, Huijuan; Chen, Jianyi; Shen, Qida; Wang, Shigui; Xu, Hongxing; Tang, Bin

    2017-01-01

    RNA interference has been used to study insects' gene function and regulation. Glycogen synthase (GS) and glycogen phosphorylase (GP) are two key enzymes in carbohydrates' conversion in insects. Glycogen content and GP and GS gene expression in several tissues and developmental stages of the Brown planthopper Nilaparvata lugens Stål (Hemiptera: Delphacidae) were analyzed in the present study, using quantitative reverse-transcription polymerase chain reaction to determine their response to double-stranded trehalases (dsTREs), trehalose-6-phosphate synthases (dsTPSs), and validamycin injection. The highest expression of both genes was detected in the wing bud, followed by leg and head tissues, and different expression patterns were shown across the developmental stages analyzed. Glycogen content significantly decreased 48 and 72 h after dsTPSs injection and 48 h after dsTREs injection. GP expression increased 48 h after dsTREs and dsTPSs injection and significantly decreased 72 h after dsTPSs, dsTRE1-1, and dsTRE1-2 injection. GS expression significantly decreased 48 h after dsTPS2 and dsTRE2 injection and 72 h after dsTRE1-1 and dsTRE1-2 injection. GP and GS expression and glycogen content significantly decreased 48 h after validamycin injection. The GP activity significantly decreased 48 h after validamycin injection, while GS activities of dsTPS1 and dsTRE2 injection groups were significantly higher than that of double-stranded GFP (dsGFP) 48 h after injection, respectively. Thus, glycogen is synthesized, released, and degraded across several insect tissues according to the need to maintain stable trehalose levels. © The Authors 2017. Published by Oxford University Press on behalf of Entomological Society of America.

  5. PENGARUH MODEL PEMBELAJARAN KOOPERATIF TIPE TPS (THINK PAIR SHARE TERHADAP PEMAHAMAN SISWA PADA MATA PELAJARAN FIQIH KELAS VII MTS AL-IRSYADIYAH DERMOLEMAHBANG SARIREJO LAMONGAN

    Directory of Open Access Journals (Sweden)

    Siti Mukhoyyaroh

    2013-05-01

    Full Text Available This paper examined some issues: 1 How is the application of cooperative learning model type TPS on Fiqh lesson; 2 How is the students’ understanding; 3 How is the effect of TPS in students’ understanding. Data collection techniques used in this paper include: 1 Observation to obtain data on the use of TPS; 2 Questionnaire to obtain data on the application of TPS and students’ understanding of Fiqh; 3 Interview and documentation was done to gain general data of the research’s object. The analyses used were: percentage and product moment. The result showed that: 1 the use of TPS was good; 2 Students’ understanding of Fiqh lesson was also good; 3 The effect of cooperative learning model type TPS in students’ understanding of Fiqh lesson, based on the result of the analysis was rxy = 0.208 with 41 respondents, while r table in the significant standard of 5% and 1% are 0.316 and 0.408. Therefore, rxy is smaller than r table which means the alternative hypothesis (Ha is ignored and zero hypothesis (Ho is accepted.

  6. SU-E-T-792: Validation of a Secondary TPS for IROC-H Recalculation of Anthropomorphic Phantoms

    Energy Technology Data Exchange (ETDEWEB)

    Kerns, J; Howell, R; Followill, D; Melancon, A; Stingo, F; Kry, S [UT MD Anderson Cancer Center, Houston, TX (United States)

    2015-06-15

    Purpose: To validate a secondary treatment planning system (sTPS) for use by the Imaging & Radiation Oncology Core-Houston (IROC-H). The TPS will recalculate phantom irradiations submitted by institutions to IROC-H and compare plan results of the institution to the sTPS. Methods: In-field dosimetric data was collected by IROC-H for numerous linacs at 6, 10, 15, and 18 MV. The data was aggregated and used to define reference linac classes; each class was then modeled in the sTPS (Mobius3D) by matching the in-field characteristics. Fields used to collect IROC-H data were recreated and recalculated using Mobius3D. The same dosimetric points were measured in the recalculation and compared to the initial collection data. Additionally, a 6MV Monte Carlo beam configuration was used to compare penumbrae in the Mobius3D models. Finally, a handful of IROC-H head and neck phantoms were recalculated using Mobius3D. Results: Recalculation and quantification of differences between reference data and Mobius3D values resulted in a relative matching score of 12.45 (0 is a perfect match) for the default 6MV Mobius3D beam configuration. By adjusting beam configuration options, iterations resulted in scores of 8.45, 6.32, and 3.52, showing that customization could have a dramatic effect on beam configuration. After in-field optimization, penumbra was compared between Monte Carlo and Mobius3D for the reference fields. For open jaw fields, FWHM field widths and penumbra widths were different by <0.6 and <1mm respectively; for MLC open fields the penumbra widths were up to 1.5mm different. Phantom recalculations showed good agreement, having an average of 0.6% error per beam. Conclusion: A secondary TPS has been validated for simple irradiation geometries using reference data collected by IROC-H. The beam was customized to the reference data iteratively and resulted in a good match. This system can provide independent recalculation of phantom plans based on independent reference data.

  7. Effective immobilization of Candida antarctica lipase B in organic-modified clays: Application for the epoxidation of terpenes

    International Nuclear Information System (INIS)

    Tzialla, Aikaterini A.; Kalogeris, Emmanuel; Enotiadis, Apostolos; Taha, Ali A.; Gournis, Dimitrios; Stamatis, Haralambos

    2009-01-01

    The use of three smectite nanoclays (Laponite, SWy-2 and Kunipia) organic-modified with octadecyl-trimethyl-ammonium surfactant, as suitable host matrices for the immobilization of lipase B from Candida antarctica (CaLB) was demonstrated. The resulting hybrid biocatalysts were characterized by a combination of powder X-ray diffraction, thermogravimetric analysis, differential thermal analysis, scanning electron microscopy and infrared spectroscopy. The experimental results confirmed the remarkable binding capacity of the three organoclays for CaLB. Activity and operational stability of immobilized CaLB were determined for the chemo-enzymatic epoxidation of terpenes (α-pinene and d-limonene) in organic media using various oxidizing agents. The immobilized enzyme retains a significant part of its activity after repeated use under drastic reaction conditions originating from the use of oxidants.

  8. Toxic effects on and structure-toxicity relationships of phenylpropanoids, terpenes, and related compounds in Aedes aegypti larvae.

    Science.gov (United States)

    Santos, Sandra R L; Silva, Viviane B; Melo, Manuela A; Barbosa, Juliana D F; Santos, Roseli L C; de Sousa, Damião P; Cavalcanti, Sócrates C H

    2010-12-01

    In the search for toxic compounds against Aedes aegypti L. (Diptera: Culicidae) larvae, a collection of commercially available aromatic and aliphatic diversely substituted compounds were selected and evaluated. p-Cymene exhibited the highest larvicidal potency LC₅₀ = 51 ppm, whereas 1,8-cineole exhibited the lowest activity value LC₅₀ = 1419 ppm. To aid future work on the search for larvicidal compounds, the structure-toxicity relationships of this collection have been evaluated. The presence of lipophilic groups results in an overall increase in potency. In general, the presence of hydroxyl groups resulted in less potent compounds. However, methylation of such hydroxyls led to an overall increase in potency. The most potent compounds showed comparably good larvicidal activity in A. aegypti larvae as other terpenes, which we assume to be the result of the increased lipophilicity.

  9. CTP synthase forms cytoophidia in the cytoplasm and nucleus

    International Nuclear Information System (INIS)

    Gou, Ke-Mian; Chang, Chia-Chun; Shen, Qing-Ji; Sung, Li-Ying; Liu, Ji-Long

    2014-01-01

    CTP synthase is an essential metabolic enzyme responsible for the de novo synthesis of CTP. Multiple studies have recently showed that CTP synthase protein molecules form filamentous structures termed cytoophidia or CTP synthase filaments in the cytoplasm of eukaryotic cells, as well as in bacteria. Here we report that CTP synthase can form cytoophidia not only in the cytoplasm, but also in the nucleus of eukaryotic cells. Both glutamine deprivation and glutamine analog treatment promote formation of cytoplasmic cytoophidia (C-cytoophidia) and nuclear cytoophidia (N-cytoophidia). N-cytoophidia are generally shorter and thinner than their cytoplasmic counterparts. In mammalian cells, both CTP synthase 1 and CTP synthase 2 can form cytoophidia. Using live imaging, we have observed that both C-cytoophidia and N-cytoophidia undergo multiple rounds of fusion upon glutamine analog treatment. Our study reveals the coexistence of cytoophidia in the cytoplasm and nucleus, therefore providing a good opportunity to investigate the intracellular compartmentation of CTP synthase. - Highlights: • CTP synthase forms cytoophidia not only in the cytoplasm but also in the nucleus. • Glutamine deprivation and Glutamine analogs promotes cytoophidium formation. • N-cytoophidia exhibit distinct morphology when compared to C-cytoophidia. • Both CTP synthase 1 and CTP synthase 2 form cytoophidia in mammalian cells. • Fusions of cytoophidia occur in the cytoplasm and nucleus

  10. CTP synthase forms cytoophidia in the cytoplasm and nucleus

    Energy Technology Data Exchange (ETDEWEB)

    Gou, Ke-Mian [MRC Functional Genomics Unit, Department of Physiology, Anatomy and Genetics, University of Oxford, Oxford OX1 3PT (United Kingdom); State Key Laboratory for Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing 100193 (China); Chang, Chia-Chun [Institute of Biotechnology, National Taiwan University, Taipei, Taiwan, ROC (China); Shen, Qing-Ji [MRC Functional Genomics Unit, Department of Physiology, Anatomy and Genetics, University of Oxford, Oxford OX1 3PT (United Kingdom); Sung, Li-Ying, E-mail: liyingsung@ntu.edu.tw [Institute of Biotechnology, National Taiwan University, Taipei, Taiwan, ROC (China); Agricultural Biotechnology Research Center, Academia Sinica, Taipei 115, Taiwan, ROC (China); Liu, Ji-Long, E-mail: jilong.liu@dpag.ox.ac.uk [MRC Functional Genomics Unit, Department of Physiology, Anatomy and Genetics, University of Oxford, Oxford OX1 3PT (United Kingdom)

    2014-04-15

    CTP synthase is an essential metabolic enzyme responsible for the de novo synthesis of CTP. Multiple studies have recently showed that CTP synthase protein molecules form filamentous structures termed cytoophidia or CTP synthase filaments in the cytoplasm of eukaryotic cells, as well as in bacteria. Here we report that CTP synthase can form cytoophidia not only in the cytoplasm, but also in the nucleus of eukaryotic cells. Both glutamine deprivation and glutamine analog treatment promote formation of cytoplasmic cytoophidia (C-cytoophidia) and nuclear cytoophidia (N-cytoophidia). N-cytoophidia are generally shorter and thinner than their cytoplasmic counterparts. In mammalian cells, both CTP synthase 1 and CTP synthase 2 can form cytoophidia. Using live imaging, we have observed that both C-cytoophidia and N-cytoophidia undergo multiple rounds of fusion upon glutamine analog treatment. Our study reveals the coexistence of cytoophidia in the cytoplasm and nucleus, therefore providing a good opportunity to investigate the intracellular compartmentation of CTP synthase. - Highlights: • CTP synthase forms cytoophidia not only in the cytoplasm but also in the nucleus. • Glutamine deprivation and Glutamine analogs promotes cytoophidium formation. • N-cytoophidia exhibit distinct morphology when compared to C-cytoophidia. • Both CTP synthase 1 and CTP synthase 2 form cytoophidia in mammalian cells. • Fusions of cytoophidia occur in the cytoplasm and nucleus.

  11. Limonene dehydrogenase hydroxylates the allylic methyl group of cyclic monoterpenes in the anaerobic terpene degradation by Castellaniella defragrans.

    Science.gov (United States)

    Puentes-Cala, Edinson; Liebeke, Manuel; Markert, Stephanie; Harder, Jens

    2018-05-01

    The enzymatic functionalization of hydrocarbons is a central step in the global carbon cycle initiating the mineralization of methane, isoprene and monoterpenes, the most abundant biologically produced hydrocarbons. Also, terpene-modifying enzymes have found many applications in the energy-economic biotechnological production of fine chemicals. Here we describe a limonene dehydrogenase that was purified from the facultatively anaerobic betaproteobacterium Castellaniella defragrans 65Phen grown on monoterpenes under denitrifying conditions in the absence of molecular oxygen. The purified limonene:ferrocenium oxidoreductase activity hydroxylated the methyl group of limonene (1-methyl-4-(1-methylethenyl)-cyclohex-1-ene) yielding perillyl alcohol ([4-(prop-1-en-2-yl)cyclohex-1-en-1-yl]methanol). The enzyme had a dithiothreitol:perillyl alcohol oxidoreductase activity yielding limonene. Mass spectrometry and molecular size determinations revealed a heterodimeric enzyme comprising CtmA and CtmB. Recently the two proteins had been identified by transposon mutagenesis and proteomics as part of the cyclic terpene metabolism ( ctm ) in Castellaniella defragrans and were annotated as FAD-dependent oxidoreductases of the protein domain family phytoene dehydrogenases and related proteins (COG1233). CtmAB is the first heterodimeric enzyme in this protein superfamily. Flavins in the purified CtmAB are oxidized by ferrocenium and are reduced by limonene. Heterologous expression of CtmA, CtmB and CtmAB in E. coli demonstrated that limonene dehydrogenase activity required both subunits carrying each a flavin cofactor. Native CtmAB oxidized a wide range of monocyclic monoterpenes containing the allylic methyl group motif (1-methyl-cyclohex-1-ene). In conclusion, we have identified CtmAB as a hydroxylating limonene dehydrogenase and the first heteromer in a family of FAD-dependent dehydrogenases acting on allylic methylene or methyl CH-bonds. We suggest a placement in EC 1

  12. Discovery and functional characterization of two diterpene synthases for sclareol biosynthesis in Salvia sclarea (L. and their relevance for perfume manufacture

    Directory of Open Access Journals (Sweden)

    Caniard Anne

    2012-07-01

    Full Text Available Abstract Background Sclareol is a diterpene natural product of high value for the fragrance industry. Its labdane carbon skeleton and its two hydroxyl groups also make it a valued starting material for semisynthesis of numerous commercial substances, including production of Ambrox® and related ambergris substitutes used in the formulation of high end perfumes. Most of the commercially-produced sclareol is derived from cultivated clary sage (Salvia sclarea and extraction of the plant material. In clary sage, sclareol mainly accumulates in essential oil-producing trichomes that densely cover flower calices. Manool also is a minor diterpene of this species and the main diterpene of related Salvia species. Results Based on previous general knowledge of diterpene biosynthesis in angiosperms, and based on mining of our recently published transcriptome database obtained by deep 454-sequencing of cDNA from clary sage calices, we cloned and functionally characterized two new diterpene synthase (diTPS enzymes for the complete biosynthesis of sclareol in clary sage. A class II diTPS (SsLPPS produced labda-13-en-8-ol diphosphate as major product from geranylgeranyl diphosphate (GGPP with some minor quantities of its non-hydroxylated analogue, (9 S, 10 S-copalyl diphosphate. A class I diTPS (SsSS then transformed these intermediates into sclareol and manool, respectively. The production of sclareol was reconstructed in vitro by combining the two recombinant diTPS enzymes with the GGPP starting substrate and in vivo by co-expression of the two proteins in yeast (Saccharomyces cerevisiae. Tobacco-based transient expression assays of green fluorescent protein-fusion constructs revealed that both enzymes possess an N-terminal signal sequence that actively targets SsLPPS and SsSS to the chloroplast, a major site of GGPP and diterpene production in plants. Conclusions SsLPPS and SsSS are two monofunctional diTPSs which, together, produce the diterpenoid

  13. Discovery and functional characterization of two diterpene synthases for sclareol biosynthesis in Salvia sclarea (L.) and their relevance for perfume manufacture.

    Science.gov (United States)

    Caniard, Anne; Zerbe, Philipp; Legrand, Sylvain; Cohade, Allison; Valot, Nadine; Magnard, Jean-Louis; Bohlmann, Jörg; Legendre, Laurent

    2012-07-26

    Sclareol is a diterpene natural product of high value for the fragrance industry. Its labdane carbon skeleton and its two hydroxyl groups also make it a valued starting material for semisynthesis of numerous commercial substances, including production of Ambrox® and related ambergris substitutes used in the formulation of high end perfumes. Most of the commercially-produced sclareol is derived from cultivated clary sage (Salvia sclarea) and extraction of the plant material. In clary sage, sclareol mainly accumulates in essential oil-producing trichomes that densely cover flower calices. Manool also is a minor diterpene of this species and the main diterpene of related Salvia species. Based on previous general knowledge of diterpene biosynthesis in angiosperms, and based on mining of our recently published transcriptome database obtained by deep 454-sequencing of cDNA from clary sage calices, we cloned and functionally characterized two new diterpene synthase (diTPS) enzymes for the complete biosynthesis of sclareol in clary sage. A class II diTPS (SsLPPS) produced labda-13-en-8-ol diphosphate as major product from geranylgeranyl diphosphate (GGPP) with some minor quantities of its non-hydroxylated analogue, (9 S, 10 S)-copalyl diphosphate. A class I diTPS (SsSS) then transformed these intermediates into sclareol and manool, respectively. The production of sclareol was reconstructed in vitro by combining the two recombinant diTPS enzymes with the GGPP starting substrate and in vivo by co-expression of the two proteins in yeast (Saccharomyces cerevisiae). Tobacco-based transient expression assays of green fluorescent protein-fusion constructs revealed that both enzymes possess an N-terminal signal sequence that actively targets SsLPPS and SsSS to the chloroplast, a major site of GGPP and diterpene production in plants. SsLPPS and SsSS are two monofunctional diTPSs which, together, produce the diterpenoid specialized metabolite sclareol in a two-step process. They

  14. The Diagnostic and Prognostic Value of Tumor Markers (CEA, SCC, CYFRA 21-1, TPS) in Head and Neck Cancer Patients.

    Science.gov (United States)

    Barak, Vivian; Meirovitz, Amichay; Leibovici, Vera; Rachmut, Jacob; Peretz, Tamar; Eliashar, Ron; Gross, Menachem

    2015-10-01

    Establishing prognostic factors is very important in the management of cancer patients. Our aim was to evaluate the clinical significance of a panel of tumor markers, including CEA (Carcino Embryonic Antigen), SCC (Squamous Cell Carcinoma Antigen), TPS (Tissue Polypeptide Specific Antigen) and CYFRA 21-1 in head and neck cancer patients, for assessing treatment response and prognosis of patients. We evaluated 312 blood samples from 143 head and neck cancer patients, from several sub-groups: 82 Larynx Carcinoma pre- and 38 post-therapy, 46 Oral Cavity pre and 29 post-therapy, 12 nasopharynx, 16 parotid and other salivary gland patients. Blood tumor markers levels were evaluated by conventional ELISA assays. Correlations of marker levels to stage of disease, lymph node involvement and therapy, were performed. Serum levels of all four tumor markers were higher before therapy and decreased thereafter in all patients. The decrease in TPS level following therapy was significant (p=0.03). Significantly higher levels of TPS and similarly higher levels of the other tumor markers were demonstrated in advanced disease (stages III and IV) patients, as opposed to early disease (stages I and II) patients (p=0.012). Node positive patients had significantly higher TPS levels as compared to node negative (p=0.02). The same trend was shown by the other markers as well, but did not reach statistical significance. TPS was best correlated to survival of patients; those having low levels had the best clinical outcome and longer survival. CEA, SCC, TPS and CYFRA 21-1 can all serve as useful tumor markers in HNC patients. They assessed response to therapy and were prognostic for recurrence. TPS proved to be the most sensitive predictor of advanced disease and poor prognosis. Copyright© 2015 International Institute of Anticancer Research (Dr. John G. Delinassios), All rights reserved.

  15. Clinical significance of Phosphatidyl Inositol Synthase overexpression in oral cancer

    International Nuclear Information System (INIS)

    Kaur, Jatinder; Sawhney, Meenakshi; DattaGupta, Siddartha; Shukla, Nootan K; Srivastava, Anurag; Ralhan, Ranju

    2010-01-01

    We reported increased levels of Phosphatidyl Inositol synthase (PI synthase), (enzyme that catalyses phosphatidyl inositol (PI) synthesis-implicated in intracellular signaling and regulation of cell growth) in smokeless tobacco (ST) exposed oral cell cultures by differential display. This study determined the clinical significance of PI synthase overexpression in oral squamous cell carcinoma (OSCC) and premalignant lesions (leukoplakia), and identified the downstream signaling proteins in PI synthase pathway that are perturbed by smokeless tobacco (ST) exposure. Tissue microarray (TMA) Immunohistochemistry, Western blotting, Confocal laser scan microscopy, RT-PCR were performed to define the expression of PI synthase in clinical samples and in oral cell culture systems. Significant increase in PI synthase immunoreactivity was observed in premalignant lesions and OSCCs as compared to oral normal tissues (p = 0.000). Further, PI synthase expression was significantly associated with de-differentiation of OSCCs, (p = 0.005) and tobacco consumption (p = 0.03, OR = 9.0). Exposure of oral cell systems to smokeless tobacco (ST) in vitro confirmed increase in PI synthase, Phosphatidylinositol 3-kinase (PI3K) and cyclin D1 levels. Collectively, increased PI synthase expression was found to be an early event in oral cancer and a target for smokeless tobacco

  16. Development of antibacterial conjugates using sulfamethoxazole with monocyclic terpenes: A systematic medicinal chemistry based computational approach.

    Science.gov (United States)

    Swain, Shasank S; Paidesetty, Sudhir K; Padhy, Rabindra N

    2017-03-01

    To develop 6 conjugate agents of the moribund antibiotic sulfamethoxazole (SMZ) joined to 6 individual monoterpenes, followed by protocols of medicinal chemistry as potent antibacterials, against multidrug resistant (MDR) human gruesome pathogenic bacteria. Antibacterial activities of the proposed conjugates were ascertained by the 'prediction of activity spectra of substances' (PASS) program. Drug-likeness parameters and toxicity profiles of conjugates were standardized with the Lipinski rule of five, using cheminformatic tools, Molsoft, molinspiration, OSIRIS and ProTox. Antibacterial activities of individual chemicals and conjugates were examined by targeting the bacterial folic acid biosynthesis enzyme, dihydropteroate synthases (DHPSs) of bacteria, Bacillus anthracis, Escherichia coli, Staphylococcus aureus, Streptococcus pneumoniae and Mycobacterium tuberculosis, with 3D structures of DHPSs from protein data bank. According to the PASS program, biological spectral values of conjugate-2, conjugate-5 and conjugate-6 were ascertained effective with 'probably active' or 'Pa' value > 0.5, for anti-infective and antituberculosic activities. Using molecular docking against 5 cited bacterial DHPSs, effective docking scores of 6 monoterpenes in the specified decreasing order (kcal/mol): -9.72 (eugenol against B. anthracis), -9.61 (eugenol against S. pneumoniae), -9. 42 (safrol, against B. anthracis), -9.39 (thymol, against M. tuberculosis), -9.34 (myristicin, against S. pneumoniae) and -9.29 (thymol, against B. anthracis); whereas the lowest docking score of SMZ was -8.46kcal/mol against S. aureus DHPS. Similarly, effective docking scores of conjugates were as specified (kcal/mol.): -10.80 (conjugate-4 consisting SMZ+safrol, against M. tuberculosis), -10.78 (conjugate-5 consisting SMZ+thymol, against M. tuberculosis), -10.60 (conjugate-5 against B. anthracis), -10.26 (conjugate-2 consisting SMZ+ eugenol, against M. tuberculosis), -10.25 (conjugate-5, against S

  17. Chrysanthemyl diphosphate synthase operates in planta as a bifunctional enzyme with chrysanthemol synthase activity

    DEFF Research Database (Denmark)

    Yang, Ting; Gao, Liping; Hu, Hao

    2014-01-01

    Chrysanthemyl diphosphate synthase (CDS) is the first path-way-specific enzyme in the biosynthesis of pyrethrins, the most widely used plant-derived pesticide. CDS catalyzes c1′-2-3 cyclopropanation reactions of two molecules of dimethylallyl diphosphate (DMAPP) to yield chrysanthemyl diphosphate...

  18. Fortifying the Bone-Implant Interface Part 1: An In Vitro Evaluation of 3D-Printed and TPS Porous Surfaces.

    Science.gov (United States)

    MacBarb, Regina F; Lindsey, Derek P; Bahney, Chelsea S; Woods, Shane A; Wolfe, Mark L; Yerby, Scott A

    2017-01-01

    An aging society and concomitant rise in the incidence of impaired bone health have led to the need for advanced osteoconductive spinal implant surfaces that promote greater biological fixation ( e.g. for interbody fusion cages, sacroiliac joint fusion implants, and artificial disc replacements). Additive manufacturing, i.e. 3D-printing, may improve bone integration by generating biomimetic spinal implant surfaces that mimic bone morphology. Such surfaces may foster an enhanced cellular response compared to traditional implant surfacing processes. This study investigated the response of human osteoblasts to additive manufactured (AM) trabecular-like titanium implant surfaces compared to traditionally machined base material with titanium plasma spray (TPS) coated surfaces, with and without a nanocrystalline hydroxyapatite (HA) coating. For TPS-coated discs, wrought Ti6Al4V ELI was machined and TPS-coating was applied. For AM discs, Ti6Al4V ELI powder was 3D-printed to form a solid base and trabecular-like porous surface. The HA-coating was applied via a precipitation dip-spin method. Surface porosity, pore size, thickness, and hydrophilicity were characterized. Initial cell attachment, proliferation, alkaline phosphatase (ALP) activity, and calcium production of hFOB cells ( n =5 per group) were measured. Cells on AM discs exhibited expedited proliferative activity. While there were no differences in mean ALP expression and calcium production between TPS and AM discs, calcium production on the AM discs trended 48% higher than on TPS discs ( p =0.07). Overall, HA-coating did not further enhance results compared to uncoated TPS and AM discs. Results demonstrate that additive manufacturing allows for controlled trabecular-like surfaces that promote earlier cell proliferation and trends toward higher calcium production than TPS coating. Results further showed that nanocrystalline HA may not provide an advantage on porous titanium surfaces. Additive manufactured porous

  19. Measurements of free and total PSA, tissue polypeptide-specific antigen (TPS), and CYFRA 21-1 in prostate cancer patients under intermittent androgen suppression therapy.

    Science.gov (United States)

    Theyer, G; Dürer, A; Theyer, U; Haberl, I; Ulsperger, E; Baumgartner, G; Hamilton, G

    1999-10-01

    The present study evaluated monthly measurements of free and total prostate-specific antigen (PSA), and the tumor proliferation markers tissue polypeptide-specific antigen (TPS) and cytokeratin fragment 21-1 (CYFRA 21-1) in patients with advanced prostate cancer receiving intermittent androgen suppression therapy (IAS). Thirty-four men received alternating cycles of 8 month androgen suppression and treatment cessation (mean duration, 10.3 months) until PSA increased to >20 microg/l. Measurements of testosterone, percentage of free PSA, TPS, and CYFRA 21-1 were performed using ELISA and RIA assays. Periods of androgen suppression resulted in reversible reductions of testosterone (from 6 +/- 0.8 to IAS cycle. TPS showed a decrease of 50% after 3 months, and CYFRA 21-1 a 25% decrease after 7 months of androgen suppression treatment. During treatment cessation, TPS exceeded the normal cutoff value of 90 U/l late in tumor regrowth (9-11 months), whereas CYFRA 21-1 remained below the normal cutoff value of 3.3 ng/ml. PSA is the best and most sensitive marker of prostate cancer regression and regrowth during IAS cycles of the markers tested in this study. Free PSA constitutes approximately 15% of total PSA (range, 5-32%), and its percentage showed no significant change during IAS cycles. The TPS and CYFRA 21-1 proliferation marker changes in IAS seem to be related mainly to effects on normal androgen-dependent tissues. Copyright 1999 Wiley-Liss, Inc.

  20. Gene Coexpression Analysis Reveals Complex Metabolism of the Monoterpene Alcohol Linalool in Arabidopsis Flowers[W][OPEN

    Science.gov (United States)

    Ginglinger, Jean-François; Boachon, Benoit; Höfer, René; Paetz, Christian; Köllner, Tobias G.; Miesch, Laurence; Lugan, Raphael; Baltenweck, Raymonde; Mutterer, Jérôme; Ullmann, Pascaline; Beran, Franziska; Claudel, Patricia; Verstappen, Francel; Fischer, Marc J.C.; Karst, Francis; Bouwmeester, Harro; Miesch, Michel; Schneider, Bernd; Gershenzon, Jonathan; Ehlting, Jürgen; Werck-Reichhart, Danièle

    2013-01-01

    The cytochrome P450 family encompasses the largest family of enzymes in plant metabolism, and the functions of many of its members in Arabidopsis thaliana are still unknown. Gene coexpression analysis pointed to two P450s that were coexpressed with two monoterpene synthases in flowers and were thus predicted to be involved in monoterpenoid metabolism. We show that all four selected genes, the two terpene synthases (TPS10 and TPS14) and the two cytochrome P450s (CYP71B31 and CYP76C3), are simultaneously expressed at anthesis, mainly in upper anther filaments and in petals. Upon transient expression in Nicotiana benthamiana, the TPS enzymes colocalize in vesicular structures associated with the plastid surface, whereas the P450 proteins were detected in the endoplasmic reticulum. Whether they were expressed in Saccharomyces cerevisiae or in N. benthamiana, the TPS enzymes formed two different enantiomers of linalool: (−)-(R)-linalool for TPS10 and (+)-(S)-linalool for TPS14. Both P450 enzymes metabolize the two linalool enantiomers to form different but overlapping sets of hydroxylated or epoxidized products. These oxygenated products are not emitted into the floral headspace, but accumulate in floral tissues as further converted or conjugated metabolites. This work reveals complex linalool metabolism in Arabidopsis flowers, the ecological role of which remains to be determined. PMID:24285789

  1. A comprehensive analysis of the Omp85/TpsB protein superfamily structural diversity, taxonomic occurrence, and evolution

    Science.gov (United States)

    Heinz, Eva; Lithgow, Trevor

    2014-01-01

    Members of the Omp85/TpsB protein superfamily are ubiquitously distributed in Gram-negative bacteria, and function in protein translocation (e.g., FhaC) or the assembly of outer membrane proteins (e.g., BamA). Several recent findings are suggestive of a further level of variation in the superfamily, including the identification of the novel membrane protein assembly factor TamA and protein translocase PlpD. To investigate the diversity and the causal evolutionary events, we undertook a comprehensive comparative sequence analysis of the Omp85/TpsB proteins. A total of 10 protein subfamilies were apparent, distinguished in their domain structure and sequence signatures. In addition to the proteins FhaC, BamA, and TamA, for which structural and functional information is available, are families of proteins with so far undescribed domain architectures linked to the Omp85 β-barrel domain. This study brings a classification structure to a dynamic protein superfamily of high interest given its essential function for Gram-negative bacteria as well as its diverse domain architecture, and we discuss several scenarios of putative functions of these so far undescribed proteins. PMID:25101071

  2. A new approach in compatibilization of the poly(lactic acid)/thermoplastic starch (PLA/TPS) blends.

    Science.gov (United States)

    Akrami, Marzieh; Ghasemi, Ismaeil; Azizi, Hamed; Karrabi, Mohammad; Seyedabadi, Mohammad

    2016-06-25

    In this study, a new compatibilizer was synthesized to improve the compatibility of the poly(lactic acid)/thermoplastic starch blends. The compatibilizer was based on maleic anhydride grafted polyethylene glycol grafted starch (mPEG-g-St), and was characterized using Fourier transform infrared spectroscopy (FTIR), dynamic mechanical thermal analysis (DMTA) and back titration techniques. The results indicated successful accomplishment of the designed reactions and formation of a starch cored structure with many connections to m-PEG chains. To assess the performance of synthesized compatibilizer, several PLA/TPS blends were prepared using an internal mixer. Consequently, their morphology, dynamic-mechanical behavior, crystallization and mechanical properties were studied. The compatibilizer enhanced interfacial adhesion, possibly due to interaction between free end carboxylic acid groups of compatibilizer and active groups of TPS and PLA phases. In addition, biodegradability of the samples was evaluated by various methods consisting of weight loss, FTIR-ATR analysis and morphology. The results revealed no considerable effect of compatibilizer on biodegradability of samples. Copyright © 2016 Elsevier Ltd. All rights reserved.

  3. Sequence analysis of cereal sucrose synthase genes and isolation ...

    African Journals Online (AJOL)

    SERVER

    2007-10-18

    Oct 18, 2007 ... sequencing of sucrose synthase gene fragment from sor- ghum using primers designed at their conserved exons. MATERIALS AND METHODS. Multiple sequence alignment. Sucrose synthase gene sequences of various cereals like rice, maize, and barley were accessed from NCBI Genbank database.

  4. Prostaglandin H synthase immunoreactivity in human gut. An immunohistochemical study

    DEFF Research Database (Denmark)

    Mikkelsen, H B; Rumessen, J J; Qvortrup, Klaus

    1991-01-01

    Prostaglandins exhibit a variety of actions on intestinal smooth muscle depending upon the type, dose and muscle layer studied. As the cellular origin of prostaglandin H (PGH) synthase has not been established with certainty in the human gut wall, we studied the localization of PGH synthase...

  5. Localization of nitric oxide synthase in human skeletal muscle

    DEFF Research Database (Denmark)

    Frandsen, Ulrik; Lopez-Figueroa, M.; Hellsten, Ylva

    1996-01-01

    The present study investigated the cellular localization of the neuronal type I and endothelial type III nitric oxide synthase in human skeletal muscle. Type I NO synthase immunoreactivity was found in the sarcolemma and the cytoplasm of all muscle fibres. Stronger immunoreactivity was expressed...

  6. Multivariate optimization of a headspace solid-phase microextraction method followed by gas chromatography with mass spectrometry for the determination of terpenes in Nicotiana langsdorffii.

    Science.gov (United States)

    Ardini, Francisco; Carro, Marina Di; Abelmoschi, Maria Luisa; Grotti, Marco; Magi, Emanuele

    2014-07-01

    A simple and sensitive procedure based on headspace solid-phase microextraction and gas chromatography with mass spectrometry was developed for the determination of five terpenes (α-pinene, limonene, linalool, α-terpineol, and geraniol) in the leaves of Nicotiana langsdorffii. The microextraction conditions (extraction temperature, equilibration time, and extraction time) were optimized by means of a Doehlert design. The experimental design showed that, for α-pinene and limonene, a low temperature and a long extraction time were needed for optimal extraction, while linalool, α-terpineol, and geraniol required a high temperature and a long extraction time. The chosen compromise conditions were temperature 60°C, equilibration time 15 min and extraction time 50 min. The main analytical figures of the optimized method were evaluated; LODs ranged from 0.07 ng/g (α-pinene) to 8.0 ng/g (geraniol), while intraday and interday repeatability were in the range 10-17% and 9-13%, respectively. Finally, the procedure was applied to in vitro wild-type and transgenic specimens of N. langsdorffii subjected to abiotic stresses (chemical and heat stress). With the exception of geraniol (75-374 ng/g), low concentration levels of terpenes were measured (ng/g level or lower); some interesting variations in terpene concentration induced by abiotic stress were observed. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  7. Inhibition of the Decrease of Volatile Esters and Terpenes During Storage of Wines and a Model Wine Medium by Wine Phenolic Extracts

    Directory of Open Access Journals (Sweden)

    Ioannis Lambropoulos

    2007-01-01

    Full Text Available The effect of red wine phenolic extracts on the stability of wine volatile esters and terpenes was examined. Muscat (white and Xinomavro (red wines were enriched with each extract at 120 or 200 mg/L, and stored in open bottles at 20 °C for 3 and 2.5 days, respectively. Moreover, a model wine medium containing isoamyl acetate, ethyl hexanoate and linalool was enriched with each extract at 100 mg/L, and stored in sealed bottles at 20 °C for 45–90 days. All samples were analysed for volatiles using SPME along with GC-MS analysis. Phenolic composition of wine extracts was determined using HPLC-DAD. No effect on the concentration of any volatiles was observed as a result of the addition of each extract in each wine or the model medium. A wine extract rich in phenolic acids and another one rich in anthocyanins and flavanols inhibited the decrease of volatile esters and terpenes in one or both wines and the model medium. Among them were several important for the aroma of wine such as ethyl acetate, isoamyl acetate, ethyl hexanoate, ethyl octanoate, ethyl decanoate and linalool. The results presented here indicate that wine phenolic acids, and anthocyanins or flavanols may be taken into account as potent inhibitors of the disappearance of volatile esters and terpenes in wines.

  8. PENGARUH STRATEGI PEMBELAJARAN READING QUESTIONING AND ANSWERING (RQA DIPADU THINK PAIR SHARE (TPS TERHADAP KETERAMPILAN METAKOGNITIF SISWA LAKI-LAKI DAN PEREMPUAN SMAN DI KOTA MALANG

    Directory of Open Access Journals (Sweden)

    Hindun Syarifah

    2016-05-01

    Full Text Available This research aimed at investigating: 1 the effect of Reading Questioning and Answering (RQA, Think Pair Share (TPS, and RQA combined with TPS learning on the students’ metacognitive skills and characters, 2 the effect of gender on students’ metacognitive skills, and 3 the effect of interaction between learning strategy and gender on students' metacognitive skills. Rancangan penelitian yang digunakan ialah quasi eksperimen dengan Pretest-Postest Nonequivalent Control Group Design .The research design used was pretest posttest nonequivalent control group design. Populasi penelitian ialah seluruh siswa kelas X SMAN Malang.The population of this research was all students of grade X State Senior High School (SSHS Malang. Sampel penelitian adalah siswa kelas X-1 SMAN 8 Malang, kelas X-6 dan X-7 SMAN 5 Malang sebagai kelas eksperimen dan siswa kelas X-3 SMAN 8 Malang sebagai kelas kontrol.The samples of this research were students of class X-1 SSHS 8 Malang, classes X-6 and X-7 SSHS 5 Malang as the experimental groups and class X-3 SSHS 8 Malang as the control group. Hasil penelitian menunjukkan hal-hal berikut: 1 Strategi pembelajaran Reading Questioning and Answering (RQA dipadu Think Pair Share (TPS berpotensi dalam memberdayakan keterampilan metakognitif siswa sebesar 17,72% lebih tinggi dibandingkan dengan strategi pembelajaran konvensional dan tidak berbeda secara signifikan dengan strategi pembelajaran RQA dan TPS.Research results showed that RQA combined with TPS learning had the potential to empower students’ metacognitive skills 17.72% higher than that of conventional learning but it was not significantly different from RQA and TPS learning. 2 Pembelajaran Think Pair Share (TPS berpotensi dalam memberdayakan karakter siswa sebesar 8,52% lebih tinggi dibandingkan dengan strategi pembelajaran konvensional, 4,16% lebih tinggi dari strategi pembelajaran RQA dipadu TPS namun tidak berbeda signifikan dari strategi pembelajaran RQA.3 Kelompok

  9. SU-C-204-01: A Fast Analytical Approach for Prompt Gamma and PET Predictions in a TPS for Proton Range Verification

    International Nuclear Information System (INIS)

    Kroniger, K; Herzog, M; Landry, G; Dedes, G; Parodi, K; Traneus, E

    2015-01-01

    Purpose: We describe and demonstrate a fast analytical tool for prompt-gamma emission prediction based on filter functions applied on the depth dose profile. We present the implementation in a treatment planning system (TPS) of the same algorithm for positron emitter distributions. Methods: The prediction of the desired observable is based on the convolution of filter functions with the depth dose profile. For both prompt-gammas and positron emitters, the results of Monte Carlo simulations (MC) are compared with those of the analytical tool. For prompt-gamma emission from inelastic proton-induced reactions, homogeneous and inhomogeneous phantoms alongside with patient data are used as irradiation targets of mono-energetic proton pencil beams. The accuracy of the tool is assessed in terms of the shape of the analytically calculated depth profiles and their absolute yields, compared to MC. For the positron emitters, the method is implemented in a research RayStation TPS and compared to MC predictions. Digital phantoms and patient data are used and positron emitter spatial density distributions are analyzed. Results: Calculated prompt-gamma profiles agree with MC within 3 % in terms of absolute yield and reproduce the correct shape. Based on an arbitrary reference material and by means of 6 filter functions (one per chemical element), profiles in any other material composed of those elements can be predicted. The TPS implemented algorithm is accurate enough to enable, via the analytically calculated positron emitters profiles, detection of range differences between the TPS and MC with errors of the order of 1–2 mm. Conclusion: The proposed analytical method predicts prompt-gamma and positron emitter profiles which generally agree with the distributions obtained by a full MC. The implementation of the tool in a TPS shows that reliable profiles can be obtained directly from the dose calculated by the TPS, without the need of full MC simulation

  10. A Comparison of the Effects of Neuronal Nitric Oxide Synthase and Inducible Nitric Oxide Synthase Inhibition on Cartilage Damage

    Directory of Open Access Journals (Sweden)

    Nevzat Selim Gokay

    2016-01-01

    Full Text Available The objective of this study was to investigate the effects of selective inducible nitric oxide synthase and neuronal nitric oxide synthase inhibitors on cartilage regeneration. The study involved 27 Wistar rats that were divided into five groups. On Day 1, both knees of 3 rats were resected and placed in a formalin solution as a control group. The remaining 24 rats were separated into 4 groups, and their right knees were surgically damaged. Depending on the groups, the rats were injected with intra-articular normal saline solution, neuronal nitric oxide synthase inhibitor 7-nitroindazole (50 mg/kg, inducible nitric oxide synthase inhibitor amino-guanidine (30 mg/kg, or nitric oxide precursor L-arginine (200 mg/kg. After 21 days, the right and left knees of the rats were resected and placed in formalin solution. The samples were histopathologically examined by a blinded evaluator and scored on 8 parameters. Although selective neuronal nitric oxide synthase inhibition exhibited significant (P=0.044 positive effects on cartilage regeneration following cartilage damage, it was determined that inducible nitric oxide synthase inhibition had no statistically significant effect on cartilage regeneration. It was observed that the nitric oxide synthase activation triggered advanced arthrosis symptoms, such as osteophyte formation. The fact that selective neuronal nitric oxide synthase inhibitors were observed to have mitigating effects on the severity of the damage may, in the future, influence the development of new agents to be used in the treatment of cartilage disorders.

  11. Chemotaxonomic significance of the terpene composition in natural populations of Pinus nigra J.F.Arnold from Serbia.

    Science.gov (United States)

    Sarac, Zorica; Bojović, Srdjan; Nikolić, Biljana; Tešević, Vele; Ethorđević, Iris; Marin, Petar D

    2013-08-01

    The essential-oil variability in seven native populations belonging to different infraspecific taxa of Pinus nigra (ssp. nigra, var. gocensis, ssp. pallasiana, and var. banatica) growing wild in Serbia was analyzed. In the needles of 195 trees from seven populations, 58 essential-oil components were identified. The major components were α-pinene (43.6%) and germacrene D (29.8%), comprising together 73.4% of the total oil composition. Based on the average chemical profile of the main terpene components (with contents >5%), the studied populations were found to be the most similar to populations from central Italy and Greece (ssp. nigra). Cluster analysis showed the division of the populations into three principal groups: the first group consisted of Populations I, II, III, IV, and V (considered as ssp. nigra group), the second of Population VI (ssp. pallasiana group), and the third of Population VII, which had the most distinct oil composition (ssp. banatica group). The taxonomic implications of the essential-oil profiles of the investigated taxa of this very complex species are discussed. Copyright © 2013 Verlag Helvetica Chimica Acta AG, Zürich.

  12. Measurements of acetone yields from the OH-initiated oxidation of terpenes by proton-transfer-reaction mass spectrometry

    International Nuclear Information System (INIS)

    Wisthaler, A.; Lindinger, W.; Jensen, N.R.; Winterhalter, R.; Hjorth, J.

    2002-01-01

    Biogenic VOCs (Volatile Organic Compounds) are known to be emitted in large quantities from vegetation exceeding largely global emissions of anthropogenic VOCs. Monoterpenes (C 10 H 16 ) are important constituents of biogenic VOC emissions. The atmospheric oxidation of Monoterpenes appears to be a potentially relevant source of acetone in the atmosphere. Acetone is present as a significant trace gas in the whole troposphere and influences in particular the atmospheric chemistry in the upper troposphere by substantially contributing to the formation of HO x radicals and peroxyacetyl nitrate (PAN). Acetone is formed promptly, following attack by the OH-radical on the terpene, via a series of highly unstable radical intermediates, but it is also formed slowly via the degradation of stable non-radical intermediates such as pinonaldehyde and nopinone. In order to investigate the relative importance of these processes, the OH-initiated oxidation of α-pinene and β-pinene was investigated in a chamber study, where the concentrations of monoterpenes, acetone, pinonaldehyde and nopinone were monitored by proton-transfer-reaction mass spectrometry (PTR-MS). It was found that significant amounts of acetone are formed directly, whenα-pinene and β-pinene are oxidized by the OH radical, but also secondary chemistry (degradation of primary reaction products) gives a significant contribution to the formation of acetone from monoterpenes. It can be concluded that atmospheric oxidation of monoterpenes contributes a significant fraction to the global acetone source strength. (nevyjel)

  13. In Vitro Cytotoxic Potential of Essential Oils of Eucalyptus benthamii and Its Related Terpenes on Tumor Cell Lines

    Directory of Open Access Journals (Sweden)

    Patrícia Mathias Döll-Boscardin

    2012-01-01

    Full Text Available Eucalyptus L. is traditionally used for many medicinal purposes. In particular, some Eucalyptus species have currently shown cytotoxic properties. Local Brazilian communities have used leaves of E. benthamii as a herbal remedy for various diseases, including cancer. Considering the lack of available data for supporting this cytotoxic effect, the goal of this paper was to study the in vitro cytotoxic potential of the essential oils from young and adult leaves of E. benthamii and some related terpenes (α-pinene, terpinen-4-ol, and γ-terpinene on Jurkat, J774A.1 and HeLa cells lines. Regarding the cytotoxic activity based on MTT assay, the essential oils showed improved results than α-pinene and γ-terpinene, particularly for Jurkat and HeLa cell lines. Terpinen-4-ol revealed a cytotoxic effect against Jurkat cells similar to that observed for volatile oils. The results of LDH activity indicated that cytotoxic activity of samples against Jurkat cells probably involved cell death by apoptosis. The decrease of cell DNA content was demonstrated due to inhibition of Jurkat cells proliferation by samples as a result of cytotoxicity. In general, the essential oils from young and adult leaves of E. benthamii presented cytotoxicity against the investigated tumor cell lines which confirms their antitumor potential.

  14. Acaricidal Potentials of the Terpene-rich Essential Oils of Two Iranian Eucalyptus Species against Tetranychus urticae Koch.

    Science.gov (United States)

    Ebadollahi, Asgar; Sendi, Jalal Jalali; Maroufpoor, Mostafa; Rahimi-Nasrabadi, Mehdi

    2017-03-01

    There is a rapid growth in the screening of plant materials for finding new bio-pesticides. In the present study, the essential oils of E. oleosa and E. torquata leaves were extracted using a Clevenger apparatus and their chemical profiles were investigated by Gas Chromatography-Mass Spectrometry (GC-MS). Among identified compounds, the terpenes had highest amount for both essential oils; 93.59% for E. oleosa and 97.69% for E. torquata. 1,8-Cineole (31.96%), α-pinene (15.25%) and trans-anethole (7.32%) in the essential oil of E. oleosa and 1,8-cineole (28.57%), α-pinene (15.74%) and globulol (13.11%) in the E. torquata essential oil were identified as the main components. The acaricidal activity of the essential oils of E. oleosa and E. torquata were examined using fumigation methods against the adult females of Tetranychus urticae Koch. The essential oils have potential acaricidal effects on T. urticae. The essential oil of E. oleosa with LC 50 value of 2.42 µL/L air was stronger than E. torquata. A correlation between log concentration and mite mortality has been observed. Based on the results of present study, it can be stated that the essential oils of E. oleosa and E. torquata have a worthy potential in the management of T. urticae.

  15. In Vitro Cytotoxic Potential of Essential Oils of Eucalyptus benthamii and Its Related Terpenes on Tumor Cell Lines

    Science.gov (United States)

    Döll-Boscardin, Patrícia Mathias; Sartoratto, Adilson; Sales Maia, Beatriz Helena Lameiro de Noronha; Padilha de Paula, Josiane; Nakashima, Tomoe; Farago, Paulo Vitor; Kanunfre, Carla Cristine

    2012-01-01

    Eucalyptus L. is traditionally used for many medicinal purposes. In particular, some Eucalyptus species have currently shown cytotoxic properties. Local Brazilian communities have used leaves of E. benthamii as a herbal remedy for various diseases, including cancer. Considering the lack of available data for supporting this cytotoxic effect, the goal of this paper was to study the in vitro cytotoxic potential of the essential oils from young and adult leaves of E. benthamii and some related terpenes (α-pinene, terpinen-4-ol, and γ-terpinene) on Jurkat, J774A.1 and HeLa cells lines. Regarding the cytotoxic activity based on MTT assay, the essential oils showed improved results than α-pinene and γ-terpinene, particularly for Jurkat and HeLa cell lines. Terpinen-4-ol revealed a cytotoxic effect against Jurkat cells similar to that observed for volatile oils. The results of LDH activity indicated that cytotoxic activity of samples against Jurkat cells probably involved cell death by apoptosis. The decrease of cell DNA content was demonstrated due to inhibition of Jurkat cells proliferation by samples as a result of cytotoxicity. In general, the essential oils from young and adult leaves of E. benthamii presented cytotoxicity against the investigated tumor cell lines which confirms their antitumor potential. PMID:22645627

  16. [Advances in metabolic engineering for the microbial production of naturally occurring terpenes-limonene and bisabolene: a mini review].

    Science.gov (United States)

    Pang, Yaru; Hu, Zhihui; Xiao, Dongguang; Yu, Aiqun

    2018-01-25

    Limonene (C₁₀H₁₆) and bisabolene (C₁₅H₂₄) are both naturally occurring terpenes in plants. Depending on the number of C₅ units, limonene and bisabolene are recognized as representative monoterpenes and sesquiterpenes, respectively. Limonene and bisabolene are important pharmaceutical and nutraceutical products used in the prevention and treatment of cancer and many other diseases. In addition, they can be used as starting materials to produce a range of commercially valuable products, such as pharmaceuticals, nutraceuticals, cosmetics, and biofuels. The low abundance or yield of limonene and bisabolene in plants renders their isolation from plant sources non-economically viable. Isolation of limonene and bisabolene from plants also suffers from low efficiency and often requires harsh reaction conditions, prolonged reaction times, and expensive equipment cost. Recently, the rapid developments in metabolic engineering of microbes provide a promising alternative route for producing these plant natural products. Therefore, producing limonene and bisabolene by engineering microbial cells into microbial factories is becoming an attractive alternative approach that can overcome the bottlenecks, making it more sustainable, environmentally friendly and economically competitive. Here, we reviewed the status of metabolic engineering of microbes that produce limonene and bisabolene including microbial hosts, key enzymes, metabolic pathways and engineering of limonene/bisabolene biosynthesis. Furthermore, key challenges and future perspectives were discussed.

  17. Homospermidine synthase, the first pathway-specific enzyme of pyrrolizidine alkaloid biosynthesis, evolved from deoxyhypusine synthase

    Science.gov (United States)

    Ober, Dietrich; Hartmann, Thomas

    1999-01-01

    Pyrrolizidine alkaloids are preformed plant defense compounds with sporadic phylogenetic distribution. They are thought to have evolved in response to the selective pressure of herbivory. The first pathway-specific intermediate of these alkaloids is the rare polyamine homospermidine, which is synthesized by homospermidine synthase (HSS). The HSS gene from Senecio vernalis was cloned and shown to be derived from the deoxyhypusine synthase (DHS) gene, which is highly conserved among all eukaryotes and archaebacteria. DHS catalyzes the first step in the activation of translation initiation factor 5A (eIF5A), which is essential for eukaryotic cell proliferation and which acts as a cofactor of the HIV-1 Rev regulatory protein. Sequence comparison provides direct evidence for the evolutionary recruitment of an essential gene of primary metabolism (DHS) for the origin of the committing step (HSS) in the biosynthesis of pyrrolizidine alkaloids. PMID:10611289

  18. Nitric oxide synthase gene G298 allele

    International Nuclear Information System (INIS)

    Nagib El-Kilany, Galal E.; Nayel, Ehab; Hazzaa, Sahar

    2004-01-01

    Background: Nitric oxide (NO) has an important effect on blood pressure, arterial wall, and the basal release of endothelial NO in hypertension (HPN) may be reduced. Until now, there is no solid data revealing the potential role of the polymorphism of the nitric oxide synthase gene (NOS) in patients with HPN and microvascular angina. Aim: The aim of the present study is to investigate the gene of endothelial nitric oxide synthase (eNOS), as the polymorphism of this gene may be a putative candidate for HPN and initiate the process of atherosclerosis. Methods: Sixty participants were recruited for this study; 50 were hypertensive patients complaining of chest pain [30 of them have electrocardiogram (EKG) changes of ischemia], 20 had isolated HPN, and 10 healthy volunteers served as control. All patients underwent stress myocardial perfusion imaging (MPI) and coronary angiography. Genotyping of eNOS for all patients and controls was performed. The linkages between HPN, microvascular angina and eNOS gene polymorphism were investigated. Results: MPI and coronary angiography revealed that 15 patients had chest pain with true ischemia and reversible myocardial perfusion defects (multiple and mild) but normal epicardial coronary arteries (microvascular angina), while 15 patients had significant coronary artery disease (CAD), and 20 hypertensive patients showed normal perfusion scan and coronary angiography. The prevalence of the NOS G 298 allele was higher in the hypertensive group with microvascular angina (documented by MPI) than it was among the control participants (P<.005). The eNOS allele was significantly higher in the hypertensive group than in the control participants, but there was no significant difference in homozygote mutants among hypertensive participants, x-syndrome and patients with CAD. Conclusion: eNOS gene polymorphism is proved to be an important etiology in microvascular angina (x-syndrome) among hypertensive patients. In addition, the eNOS mutant

  19. Glycogen synthase activation by sugars in isolated hepatocytes.

    Science.gov (United States)

    Ciudad, C J; Carabaza, A; Bosch, F; Gòmez I Foix, A M; Guinovart, J J

    1988-07-01

    We have investigated the activation by sugars of glycogen synthase in relation to (i) phosphorylase a activity and (ii) changes in the intracellular concentration of glucose 6-phosphate and adenine nucleotides. All the sugars tested in this work present the common denominator of activating glycogen synthase. On the other hand, phosphorylase a activity is decreased by mannose and glucose, unchanged by galactose and xylitol, and increased by tagatose, glyceraldehyde, and fructose. Dihydroxyacetone exerts a biphasic effect on phosphorylase. These findings provide additional evidence proving that glycogen synthase can be activated regardless of the levels of phosphorylase a, clearly establishing that a nonsequential mechanism for the activation of glycogen synthase occurs in liver cells. The glycogen synthase activation state is related to the concentrations of glucose 6-phosphate and adenine nucleotides. In this respect, tagatose, glyceraldehyde, and fructose deplete ATP and increase AMP contents, whereas glucose, mannose, galactose, xylitol, and dihydroxyacetone do not alter the concentration of these nucleotides. In addition, all these sugars, except glyceraldehyde, increase the intracellular content of glucose 6-phosphate. The activation of glycogen synthase by sugars is reflected in decreases on both kinetic constants of the enzyme, M0.5 (for glucose 6-phosphate) and S0.5 (for UDP-glucose). We propose that hepatocyte glycogen synthase is activated by monosaccharides by a mechanism triggered by changes in glucose 6-phosphate and adenine nucleotide concentrations which have been described to modify glycogen synthase phosphatase activity. This mechanism represents a metabolite control of the sugar-induced activation of hepatocyte glycogen synthase.

  20. Kepler Planet Detection Metrics: Per-Target Flux-Level Transit Injection Tests of TPS for Data Release 25

    Science.gov (United States)

    Burke, Christopher J.; Catanzarite, Joseph

    2017-01-01

    Quantifying the ability of a transiting planet survey to recover transit signals has commonly been accomplished through Monte-Carlo injection of transit signals into the observed data and subsequent running of the signal search algorithm (Gilliland et al., 2000; Weldrake et al., 2005; Burke et al., 2006). In order to characterize the performance of the Kepler pipeline (Twicken et al., 2016; Jenkins et al., 2017) on a sample of over 200,000 stars, two complementary injection and recovery tests are utilized:1. Injection of a single transit signal per target into the image or pixel-level data, hereafter referred to as pixel-level transit injection (PLTI), with subsequent processing through the Photometric Analysis (PA), Presearch Data Conditioning (PDC), Transiting Planet Search (TPS), and Data Validation (DV) modules of the Kepler pipeline. The PLTI quantification of the Kepler pipeline's completeness has been described previously by Christiansen et al. (2015, 2016); the completeness of the final SOC 9.3 Kepler pipeline acting on the Data Release 25 (DR25) light curves is described by Christiansen (2017).2. Injection of multiple transit signals per target into the normalized flux time series data with a subsequent transit search using a stream-lined version of the Transiting Planet Search (TPS) module. This test, hereafter referred to as flux-level transit injection (FLTI), is the subject of this document. By running a heavily modified version of TPS, FLTI is able to perform many injections on selected targets and determine in some detail which injected signals are recoverable. Significant numerical efficiency gains are enabled by precomputing the data conditioning steps at the onset of TPS and limiting the search parameter space (i.e., orbital period, transit duration, and ephemeris zero-point) to a small region around each injected transit signal.The PLTI test has the advantage that it follows transit signals through all processing steps of the Kepler pipeline, and

  1. SU-F-T-49: Dosimetry Parameters and TPS Commissioning for the CivaSheet Directional Pd-103 Brachytherapy Source

    Energy Technology Data Exchange (ETDEWEB)

    Rivard, MJ [Tufts University School of Medicine, Boston, MA (United States)

    2016-06-15

    Purpose: The CivaSheet is a new LDR Pd-103 brachytherapy device offering directional-radiation for preferentially irradiating malignancies with healthy-tissue sparing. Observations are presented on dosimetric characterization, TPS commissioning, and evaluation of the dosesuperposition- principle for summing individual elements comprising a planar CivaSheet Methods: The CivaSheet comprises individual sources (CivaDots, 0.05cm thick and 0.25cm diam.) inside a flexible bioabsorbable substrate with a 0.8cm center-to-center rectangular array. All non-radioactive components were measured to ensure accuracy of manufacturer-provided dimensional information. The Pd spatial distribution was gleaned from radioactive and inert samples, then modeled with the MCNP6 radiation-transport-code. A 6×6 array CivaSheet was modeled to evaluate the dose superposition principle for treatment planning. Air-kerma-strength was estimated using the NIST WAFAC geometry. Absorbed dose was estimated in water with polar sampling covering 0.05≤r≤15cm in 0.05cm increments and 0°≤θ≤180° in 1° increments. These data were entered into VariSeed9.0 and tested for the dose-superposition-principle. Results: The dose-rate-constant was 0.579 cGy/h/U with g(r) determined along the rotational-axis of symmetry (0°) instead of 90°. gP(r) values at 0.1, 0.5, 2, 5, and 10cm were 1.884, 1.344, 0.558, 0.088, and 0.0046. F(r,θ) decreased between 0° and 180° by factors of 270, 23, and 5.1 at 0.1, 1, and 10cm. The highest dose-gradient was at 92°, changing by a factor of 3 within 1° due to Au-foil shielding. TPS commissioning from 0.1≤r≤11cm and 0°≤θ≤180° demonstrated 2% reproducibility of input data except at the high-dose-gradient where interpolations caused 3% differences. Dose superposition of CivaDots replicated a multi-source CivaSheet array within 2% except where another CivaDot was present. Following implantation, the device is not perfectly planar. TPS accuracy utilizing the dose

  2. Generation and Functional Evaluation of Designer Monoterpene Synthases.

    Science.gov (United States)

    Srividya, N; Lange, I; Lange, B M

    2016-01-01

    Monoterpene synthases are highly versatile enzymes that catalyze the first committed step in the pathways toward terpenoids, the structurally most diverse class of plant natural products. Recent advancements in our understanding of the reaction mechanism have enabled engineering approaches to develop mutant monoterpene synthases that produce specific monoterpenes. In this chapter, we are describing protocols to introduce targeted mutations, express mutant enzyme catalysts in heterologous hosts, and assess their catalytic properties. Mutant monoterpene synthases have the potential to contribute significantly to synthetic biology efforts aimed at producing larger amounts of commercially attractive monoterpenes. © 2016 Elsevier Inc. All rights reserved.

  3. Geranylfarnesyl diphosphate synthase from Methanosarcina mazei: Different role, different evolution

    International Nuclear Information System (INIS)

    Ogawa, Takuya; Yoshimura, Tohru; Hemmi, Hisashi

    2010-01-01

    The gene of (all-E) geranylfarnesyl diphosphate synthase that is responsible for the biosynthesis of methanophenazine, an electron carrier utilized for methanogenesis, was cloned from a methanogenic archaeon Methanosarcina mazei Goe1. The properties of the recombinant enzyme and the results of phylogenetic analysis suggest that the enzyme is closely related to (all-E) prenyl diphosphate synthases that are responsible for the biosynthesis of respiratory quinones, rather than to the enzymes involved in the biosynthesis of archaeal membrane lipids, including (all-E) geranylfarnesyl diphosphate synthase from a thermophilic archaeon.

  4. SU-F-T-94: Plan2pdf - a Software Tool for Automatic Plan Report for Philips Pinnacle TPS

    Energy Technology Data Exchange (ETDEWEB)

    Wu, C [Sutter Medical Foundation, Roseville, CA (United States)

    2016-06-15

    Purpose: To implement an automatic electronic PDF plan reporting tool for Philips Pinnacle treatment planning system (TPS) Methods: An electronic treatment plan reporting software is developed by us to enable fully automatic PDF report from Pinnacle TPS to external EMR programs such as MOSAIQ. The tool is named “plan2pdf”. plan2pdf is implemented using Pinnacle scripts, Java and UNIX shell scripts, without any external program needed. plan2pdf supports full auto-mode and manual mode reporting. In full auto-mode, with a single mouse click, plan2pdf will generate a detailed Pinnacle plan report in PDF format, which includes customizable cover page, Pinnacle plan summary, orthogonal views through each plan POI and maximum dose point, DRR for each beam, serial transverse views captured throughout the dose grid at a user specified interval, DVH and scorecard windows. The final PDF report is also automatically bookmarked for each section above for convenient plan review. The final PDF report can either be saved on a user specified folder on Pinnacle, or it can be automatically exported to an EMR import folder via a user configured FTP service. In manual capture mode, plan2pdf allows users to capture any Pinnacle plan by full screen, individual window or rectangular ROI drawn on screen. Furthermore, to avoid possible patients’ plan mix-up during auto-mode reporting, a user conflict check feature is included in plan2pdf: it prompts user to wait if another patient is being exported by plan2pdf by another user. Results: plan2pdf is tested extensively and successfully at our institution consists of 5 centers, 15 dosimetrists and 10 physicists, running Pinnacle version 9.10 on Enterprise servers. Conclusion: plan2pdf provides a highly efficient, user friendly and clinical proven platform for all Philips Pinnacle users, to generate a detailed plan report in PDF format for external EMR systems.

  5. SU-F-T-94: Plan2pdf - a Software Tool for Automatic Plan Report for Philips Pinnacle TPS

    International Nuclear Information System (INIS)

    Wu, C

    2016-01-01

    Purpose: To implement an automatic electronic PDF plan reporting tool for Philips Pinnacle treatment planning system (TPS) Methods: An electronic treatment plan reporting software is developed by us to enable fully automatic PDF report from Pinnacle TPS to external EMR programs such as MOSAIQ. The tool is named “plan2pdf”. plan2pdf is implemented using Pinnacle scripts, Java and UNIX shell scripts, without any external program needed. plan2pdf supports full auto-mode and manual mode reporting. In full auto-mode, with a single mouse click, plan2pdf will generate a detailed Pinnacle plan report in PDF format, which includes customizable cover page, Pinnacle plan summary, orthogonal views through each plan POI and maximum dose point, DRR for each beam, serial transverse views captured throughout the dose grid at a user specified interval, DVH and scorecard windows. The final PDF report is also automatically bookmarked for each section above for convenient plan review. The final PDF report can either be saved on a user specified folder on Pinnacle, or it can be automatically exported to an EMR import folder via a user configured FTP service. In manual capture mode, plan2pdf allows users to capture any Pinnacle plan by full screen, individual window or rectangular ROI drawn on screen. Furthermore, to avoid possible patients’ plan mix-up during auto-mode reporting, a user conflict check feature is included in plan2pdf: it prompts user to wait if another patient is being exported by plan2pdf by another user. Results: plan2pdf is tested extensively and successfully at our institution consists of 5 centers, 15 dosimetrists and 10 physicists, running Pinnacle version 9.10 on Enterprise servers. Conclusion: plan2pdf provides a highly efficient, user friendly and clinical proven platform for all Philips Pinnacle users, to generate a detailed plan report in PDF format for external EMR systems.

  6. Discovery of germacrene A synthases in Barnadesia spinosa: The first committed step in sesquiterpene lactone biosynthesis in the basal member of the Asteraceae.

    Science.gov (United States)

    Nguyen, Trinh-Don; Faraldos, Juan A; Vardakou, Maria; Salmon, Melissa; O'Maille, Paul E; Ro, Dae-Kyun

    2016-10-28

    The Andes-endemic Barnadesioideae lineage is the oldest surviving and phylogenetically basal subfamily of the Asteraceae (Compositae), a prolific group of flowering plants with world-wide distribution (∼24,000 species) marked by a rich diversity of sesquiterpene lactones (STLs). Intriguingly, there is no evidence that members of the Barnadesioideae produce STLs, specialized metabolites thought to have contributed to the adaptive success of the Asteraceae family outside South America. The biosynthesis of STLs requires the intimate expression and functional integration of germacrene A synthase (GAS) and germacrene A oxidase (GAO) to sequentially cyclize and oxidize farnesyl diphosphate into the advanced intermediate germacrene A acid leading to diverse STLs. Our previous discovery of GAO activity conserved across all major subfamilies of Asteraceae, including the phylogenetically basal lineage of Barnadesioideae, prompted further investigation of the presence of the gateway GAS in Barnadesioideae. Herein we isolated two terpene synthases (BsGAS1/BsGAS2) from the basal Barnadesia spinosa (Barnadesioideae) that displayed robust GAS activity when reconstituted in yeast and characterized in vitro. Despite the apparent lack of STLs in the Barnadesioideae, this work unambiguously confirms the presence of GAS in the basal genera of the Asteraceae. Phylogenetic analysis reveals that the two BsGASs fall into two distinct clades of the Asteraceae's GASs, and BsGAS1 clade is only retained in the evolutionary closer Cichorioideae subfamily, implicating BsGAS2 is likely the ancestral base of most GASs found in the lineages outside the Barnadesioideae. Taken together, these results show the enzymatic capacities of GAS and GAO emerged prior to the subsequent radiation of STL-producing Asteraceae subfamilies. Copyright © 2016 Elsevier Inc. All rights reserved.

  7. Synthesis and characterization of the NiFe{sub 2}O{sub 4}@TEOS–TPS@Ag nanocomposite and investigation of its antibacterial activity

    Energy Technology Data Exchange (ETDEWEB)

    Allafchian, Ali R., E-mail: Allafchian@cc.iut.ac.ir [Nanotechnology and Advanced Materials Institute, Isfahan University of Technology, Isfahan 84156-83111 (Iran, Islamic Republic of); Jalali, S.A.H., E-mail: ahjalali2002@gmail.com [Department of Natural Resources, Isfahan University of Technology, Isfahan 84156-83111 (Iran, Islamic Republic of); Institute of Biotechnology and Bioengineering, Isfahan University of Technology, Isfahan 84156-83111 (Iran, Islamic Republic of); Amiri, R., E-mail: razieh.amiri@gmail.com [Nanotechnology and Advanced Materials Institute, Isfahan University of Technology, Isfahan 84156-83111 (Iran, Islamic Republic of); Shahabadi, Sh., E-mail: shirinshahabadi@rocketmail.com [Nanotechnology and Advanced Materials Institute, Isfahan University of Technology, Isfahan 84156-83111 (Iran, Islamic Republic of)

    2016-11-01

    Highlights: • The new NiFe{sub 2}O{sub 4}@TEOS–TPS@Ag nanocomposite was synthesized and characterized. • The VSM technique was applied for investigation of their magnetic properties. • By using magnetic decantation, they can be easily removed from the disinfected media. • These nanocomposites exhibit good antibacterial activity and high reusability. - Abstract: In this study, the NiFe{sub 2}O{sub 4} was embedded in (3–mercaptopropyl) trimethoxysilane (TPS) and tetraethyl orthosilicate (TEOS) using the sol–gel method. These compounds were used as the support of Ag nanoparticles (Ag NPs). The NiFe{sub 2}O{sub 4}@TEOS–TPS@Ag nanocomposites were obtained with the development of bonding between the silver atoms of Ag NPs and the sulfur atoms of TPS molecule. Field emission scanning electron microscopy (FE–SEM), transmission electron microscopy (TEM), X–ray diffraction (XRD) and Fourier transform infrared spectroscopy (FT–IR) were used for the characterization of the Ag nanocomposites. Also, the magnetic properties of these nanocomposites were studied by using a vibrating sample magnetometer (VSM) technique. The disk diffusion, minimum inhibition concentration (MIC) and minimum bactericidal concentrations (MBC) tests were used for the investigation of the antibacterial effect of this nanocomposite against bacterial strains. The synthesized nanocomposite presented high reusability and good antibacterial activity against gram–positive and gram–negative bacteria. Remarkably, this nanocomposite could be easily removed from the disinfected media by magnetic decantation.

  8. Comparison effectiveness of cooperative learning type STAD with cooperative learning type TPS in terms of mathematical method of Junior High School students

    Science.gov (United States)

    Wahyuni, A.

    2018-05-01

    This research is aimed to find out whether the model of cooperative learning type Student Team Achievement Division (STAD) is more effective than cooperative learning type Think-Pair-Share in SMP Negeri 7 Yogyakarta. This research was a quasi-experimental research, using two experimental groups. The population of research was all students of 7thclass in SMP Negeri 7 Yogyakarta that consists of 5 Classes. From the population were taken 2 classes randomly which used as sample. The instrument to collect data was a description test. Measurement of instrument validity use content validity and construct validity, while measuring instrument reliability use Cronbach Alpha formula. To investigate the effectiveness of cooperative learning type STAD and cooperative learning type TPS on the aspect of student’s mathematical method, the datas were analyzed by one sample test. Comparing the effectiveness of cooperative learning type STAD and TPS in terms of mathematical communication skills by using t-test. Normality test was not conducted because the sample of research more than 30 students, while homogeneity tested by using Kolmogorov Smirnov test. The analysis was performed at 5% confidence level.The results show as follows : 1) The model of cooperative learning type STAD and TPS are effective in terms of mathematical method of junior high school students. 2). STAD type cooperative learning model is more effective than TPS type cooperative learning model in terms of mathematical methods of junior high school students.

  9. The effect of maleinized linseed oil (MLO) on mechanical performance of poly(lactic acid)-thermoplastic starch (PLA-TPS) blends.

    Science.gov (United States)

    Ferri, J M; Garcia-Garcia, D; Sánchez-Nacher, L; Fenollar, O; Balart, R

    2016-08-20

    In this work, poly(lactic acid), PLA and thermoplastic starch, TPS blends (with a fixed content of 30wt.% TPS) were prepared by melt extrusion process to increase the low ductile properties of PLA. The TPS used contains an aliphatic/aromatic biodegradable polyester (AAPE) that provides good resistance to aging and moisture. This blend provides slightly improved ductile properties with an increase in elongation at break of 21.5% but phase separation is observed due to the lack of strong interactions between the two polymers. Small amounts of maleinized linseed oil (MLO) can positively contribute to improve the ductile properties of these blends by a combined plasticizing-compatibilizing effect. The elongation at break increases over 160% with the only addition of 6phr MLO. One of the evidence of the plasticizing-compatibilizing effect provided by MLO is the change in the glass transition temperature (Tg) with a decrease of about 10°C. Field emission scanning electron microscopy (FESEM) of PLA-TPS blends with varying amounts of maleinized linseed oil also suggests an increase in compatibility. Copyright © 2016 Elsevier Ltd. All rights reserved.

  10. Morphological and mechanical properties of thermoplastic starch (TPS) and its blend with poly(lactic acid)(PLA) using cassava bagasse and starch

    International Nuclear Information System (INIS)

    Teixeira, Eliangela de M.; Correa, Ana C.; Campos, Adriana de; Marconcini, Jose M.; Mattoso, Luiz H.C.; Curvelo, Antonio A.S.

    2011-01-01

    This study aims the use of an agro waste coming from the industrialization of cassava starch, known as cassava bagasse (BG). This material contains residual starch and cellulose fibers which can be used to obtain thermoplastic starch (TPS) and /or blends reinforced with fibers. In this context, it was prepared a thermoplastic starch with BG (TPSBG) and evaluated the incorporation of 20wt% of it into the biodegradable polymer poly (lactic acid) (PLA), resulting in a blend PLA/TPSBG20. The materials were investigated through morphology (scanning electron microscopy with field emission gun (FEG), x-ray diffraction (XRD), and mechanical behavior (tensile test). Their properties were compared to the blend PLA/TPSI20 in which TPSI is obtained from commercial cassava starch. The results showed that the use of bagasse generates homogenous materials with higher mechanical strength if compared to TPS obtained from commercial cassava starch. The fiber in this residue acted as reinforcement for TPS and PLA/TPS systems. (author)

  11. CARACTERIZACIÓN FÍSICO-MECÁNICA DE UN ALMIDÓN TERMOPLÁSTICO (TPS DE YUCA Y ANÁLISIS INTERFACIAL CON FIBRAS DE FIQUE CARACTERIZAÇÃO FÍSICO-MECÂNICO DE UM AMIDO TERMOPLÁSTICO (TPS DE MANDIOCA E ANÁLISE INTERFACIAL DE FIBRAS DE FIQUE PHYSICO-MECHANICAL CHARACTERIZATION OF THERMOPLASTIC CASSAVA STARCH (TPS AND INTERFACIAL ANALYSIS WITH FIQUE FIBERS

    Directory of Open Access Journals (Sweden)

    JOSE H. MINA H.

    2012-12-01

    Full Text Available Con el fin de estimar la factibilidad de reforzar una matriz de almidón termoplástico (TPS con fibras de Fique, se efectuaron pruebas de micro-mecánica mediante la técnica de zafado de una fibra (pull-out. El TPS se obtuvo a partir de la plastificación, con glicerol, de un almidón nativo de yuca (Manihot sculenta Crantz, empleando un proceso de extrusión. Para estimar la resistencia cortante interfacial (RCI se utilizaron tanto fibras de Fique nativas como modificadas superficialmente, a partir de un tratamiento alcalino. Para la preparación de los especímenes de ensayo el TPS fue peletizado, molido y posteriormente conformado en forma de placas siguiendo un proceso de moldeo por compresión; el material fue acondicionado a 25°C y 54% de humedad relativa, antes de su caracterización físico-química y mecánica. Se encontró que el TPS presentó una absorción de humedad en el equilibrio del orden del 10.5% y resistencia máxima, módulo de elasticidad y deformación a tensión de 1.5 MPa, 42.5 MPa y 26%, respectivamente. Por otro lado, se llegó a que la resistencia cortante interfacial (RCI entre el TPS y las fibras aumentó con el tratamiento superficial del Fique, debiéndose esto al aumento de rugosidad alcanzado con la alcalinización.A fim de estimar-se a viabilidade de reforçar a matriz de amido termoplástico (TPS com fibras de Fique, foram testados usando técnica de micro-mecânico (pull-out. TPS foi obtido a partir de plastificação, com o glicerol, um amido nativo de mandioca (manihot sculenta Crantz utilizando um processo de extrusão. Para estimar a resistência ao cisalhamento interfacial (RCI foram utilizadas fibras nativas e fique superfície modificada, a partir de um tratamento alcalino. Para a preparação das amostras de teste TPS foi peletizado, moído e, em seguida, moldagem sob a forma de placas como um processo de moldagem por compressão, o material foi condicionado a 25°C e 54% umidade relativa HR, antes

  12. The volatile profiles of a rare apple (Malus domestica Borkh.) honey: shikimic acid-pathway derivatives, terpenes, and others.

    Science.gov (United States)

    Kuś, Piotr Marek; Jerković, Igor; Tuberoso, Carlo Ignazio Giovanni; Šarolić, Mladenka

    2013-09-01

    The volatile profiles of rare Malus domestica Borkh. honey were investigated for the first time. Two representative samples from Poland (sample I) and Spain (sample II) were selected by pollen analysis (44-45% of Malus spp. pollen) and investigated by GC/FID/MS after headspace solid-phase microextraction (HS-SPME) and ultrasonic solvent extraction (USE). The apple honey is characterized by high percentage of shikimic acid-pathway derivatives, as well as terpenes, norisoprenoids, and some other compounds such as coumaran and methyl 1H-indole-3-acetate. The main compounds of the honey headspace were (sample I; sample II): benzaldehyde (9.4%; 32.1%), benzyl alcohol (0.3%; 14.4%), hotrienol (26.0%, 6.2%), and lilac aldehyde isomers (26.3%; 1.7%), but only Spanish sample contained car-2-en-4-one (10.2%). CH2 Cl2 and pentane/Et2 O 1 : 2 (v/v) were used for USE. The most relevant compounds identified in the extracts were: benzaldehyde (0.9-3.9%), benzoic acid (2.0-11.2%), terpendiol I (0.3-7.4%), coumaran (0.0-2.8%), 2-phenylacetic acid (2.0-26.4%), methyl syringate (3.9-13.1%), vomifoliol (5.0-31.8%), and methyl 1H-indole-3-acetate (1.9-10.2%). Apple honey contained also benzyl alcohol, 2-phenylethanol, (E)-cinnamaldehyde, (E)-cinnamyl alcohol, eugenol, vanillin, and linalool that have been found previously in apple flowers, thus disclosing similarity of both volatile profiles. Copyright © 2013 Verlag Helvetica Chimica Acta AG, Zürich.

  13. Isolation of developing secondary xylem specific cellulose synthase ...

    Indian Academy of Sciences (India)

    The present study aimed at identifying developing secondary xylem specific cellulose synthase genes from .... the First strand cDNA synthesis kit (Fermentas, Pittsburgh,. USA). .... ing height of the rooted cutting, girth of the stem, leaf area.

  14. Cloning and expression of pineapple sucrose- phosphate synthase ...

    African Journals Online (AJOL)

    hope&shola

    2010-12-06

    Dec 6, 2010 ... phosphate; EDTA, ethylene diamine tetraacetic acid; Ivr, invertase; SS .... phenolics, tannins and artifacts due to differences of tissue composition ..... Banana sucrose-phosphate synthase gene expression during fruit ripening.

  15. Modulation of hyaluronan synthase activity in cellular membrane fractions

    OpenAIRE

    Vigetti, Davide; Genasetti, A; Karousou, Evgenia; Viola, Manuela; Clerici, M; Bartolini, B; Moretto, Paola; DE LUCA, Giancarlo; Hascall, Vc; Passi, Alberto

    2009-01-01

    Hyaluronan (HA), the only non-sulfated glycosaminoglycan, is involved in morphogenesis, wound healing, inflammation, angiogenesis, and cancer. In mammals, HA is synthesized by three homologous HA synthases, HAS1, HAS2, and HAS3, that polymerize the HA chain using UDP-glucuronic acid and UDP-N-acetylglucosamine as precursors. Since the amount of HA is critical in several pathophysiological conditions, we developed a non-radioactive assay for measuring the activity of HA synthases (HASs) in euk...

  16. Inhibitors of Fatty Acid Synthase for Prostate Cancer. Revision

    Science.gov (United States)

    2013-05-01

    acetyl- cholinesterase inhibitors have been developed, many with femtomolar binding affinities (7). This body of literature also confirms that the...AD_________________ Award Number: W81XWH-09-1-0204 TITLE: Inhibitors of Fatty Acid Synthase for...May 2013 2. REPORT TYPE Revised Final 3. DATES COVERED 01 May 2009-30 Apr 2013 4. TITLE AND SUBTITLE Inhibitors of Fatty Acid Synthase for

  17. Inhibitors of Fatty Acid Synthase for Prostate Cancer

    Science.gov (United States)

    2012-05-01

    compounds. For example, numerous classes of acetyl- cholinesterase inhibitors have been developed, m any with fe mtomolar binding affinities (7). This...AD_________________ Award Number: W81XWH-09-1-0204 TITLE: Inhibitors of Fatty Acid Synthase for...CONTRACT NUMBER Inhibitors of Fatty Acid Synthase for Prostate Cancer 5b. GRANT NUMBER W81XWH-09-1-0204 5c. PROGRAM ELEMENT NUMBER 6. AUTHOR

  18. Homocystinuria due to cystathionine beta synthase deficiency

    Directory of Open Access Journals (Sweden)

    Rao T

    2008-01-01

    Full Text Available A two year-old male child presented with cutis marmorata congenita universalis, brittle hair, mild mental retardation, and finger spasms. Biochemical findings include increased levels of homocysteine in the blood-106.62 µmol/L (normal levels: 5.90-16µmol/L. Biochemical tests such as the silver nitroprusside and nitroprusside tests were positive suggesting homocystinuria. The patient was treated with oral pyridoxine therapy for three months. The child responded well to this therapy and the muscle spasms as well as skin manifestations such as cutis marmorata subsided. The treatment is being continued; the case is reported here because of its rarity. Homocysteinuria arising due to cystathionine beta-synthase (CBS deficiency is an autosomal recessive disorder of methionine metabolism that produces increased levels of urinary homocysteine and methionine It manifests itself in vascular, central nervous system, cutaneous, and connective tissue disturbances and phenotypically resembles Marfan′s syndrome. Skin manifestations include malar flush, thin hair, and cutis reticulata / marmorata.

  19. Sphingomyelin synthases regulate protein trafficking and secretion.

    Directory of Open Access Journals (Sweden)

    Marimuthu Subathra

    Full Text Available Sphingomyelin synthases (SMS1 and 2 represent a class of enzymes that transfer a phosphocholine moiety from phosphatidylcholine onto ceramide thus producing sphingomyelin and diacylglycerol (DAG. SMS1 localizes at the Golgi while SMS2 localizes both at the Golgi and the plasma membrane. Previous studies from our laboratory showed that modulation of SMS1 and, to a lesser extent, of SMS2 affected the formation of DAG at the Golgi apparatus. As a consequence, down-regulation of SMS1 and SMS2 reduced the localization of the DAG-binding protein, protein kinase D (PKD, to the Golgi. Since PKD recruitment to the Golgi has been implicated in cellular secretion through the trans golgi network (TGN, the effect of down-regulation of SMSs on TGN-to-plasma membrane trafficking was studied. Down regulation of either SMS1 or SMS2 significantly retarded trafficking of the reporter protein vesicular stomatitis virus G protein tagged with GFP (VSVG-GFP from the TGN to the cell surface. Inhibition of SMSs also induced tubular protrusions from the trans Golgi network reminiscent of inhibited TGN membrane fission. Since a recent study demonstrated the requirement of PKD activity for insulin secretion in beta cells, we tested the function of SMS in this model. Inhibition of SMS significantly reduced insulin secretion in rat INS-1 cells. Taken together these results provide the first direct evidence that both enzymes (SMS1 and 2 are capable of regulating TGN-mediated protein trafficking and secretion, functions that are compatible with PKD being a down-stream target for SMSs in the Golgi.

  20. Terpenes tell different tales at different scales: glimpses into the Chemical Ecology of conifer - bark beetle - microbial interactions.

    Science.gov (United States)

    Raffa, Kenneth F

    2014-01-01

    , biochemical time), relationships among inducible and constitutive defenses, population dynamics, and plastic host-selection behavior (stand level, ecological time), and climate-driven range expansion of a native eruptive species into semi-naïve and potentially naïve habitats (geographical level, evolutionary time). I approach this problem by focusing primarily on one chemical group, terpenes, by emphasizing the curvilinear and threshold-structured basis of most underlying relationships, and by focusing on the system's feedback structure, which can either buffer or amplify relationships across scales.

  1. Characterization of the human gene (TBXAS1) encoding thromboxane synthase.

    Science.gov (United States)

    Miyata, A; Yokoyama, C; Ihara, H; Bandoh, S; Takeda, O; Takahashi, E; Tanabe, T

    1994-09-01

    The gene encoding human thromboxane synthase (TBXAS1) was isolated from a human EMBL3 genomic library using human platelet thromboxane synthase cDNA as a probe. Nucleotide sequencing revealed that the human thromboxane synthase gene spans more than 75 kb and consists of 13 exons and 12 introns, of which the splice donor and acceptor sites conform to the GT/AG rule. The exon-intron boundaries of the thromboxane synthase gene were similar to those of the human cytochrome P450 nifedipine oxidase gene (CYP3A4) except for introns 9 and 10, although the primary sequences of these enzymes exhibited 35.8% identity each other. The 1.2-kb of the 5'-flanking region sequence contained potential binding sites for several transcription factors (AP-1, AP-2, GATA-1, CCAAT box, xenobiotic-response element, PEA-3, LF-A1, myb, basic transcription element and cAMP-response element). Primer-extension analysis indicated the multiple transcription-start sites, and the major start site was identified as an adenine residue located 142 bases upstream of the translation-initiation site. However, neither a typical TATA box nor a typical CAAT box is found within the 100-b upstream of the translation-initiation site. Southern-blot analysis revealed the presence of one copy of the thromboxane synthase gene per haploid genome. Furthermore, a fluorescence in situ hybridization study revealed that the human gene for thromboxane synthase is localized to band q33-q34 of the long arm of chromosome 7. A tissue-distribution study demonstrated that thromboxane synthase mRNA is widely expressed in human tissues and is particularly abundant in peripheral blood leukocyte, spleen, lung and liver. The low but significant levels of mRNA were observed in kidney, placenta and thymus.

  2. Preparação, caracterização e degradação de blendas PS/TPS usando glicerol e óleo de buriti como plastificantes Preparation, characterization and degradation of PS/TPS blends using glycerol and buriti oil as plastiscizers

    Directory of Open Access Journals (Sweden)

    Daniela Schlemmer

    2010-01-01

    Full Text Available Poliestireno (PS é um dos materiais descartáveis mais usados no nosso cotidiano, mas ele não se decompõe facilmente. A adição de amido de mandioca ao PS pode melhorar a sua degradabilidade, tendo em vista ser o amido um polímero biodegradável. O objetivo deste trabalho foi desenvolver blendas de PS e (amido termoplástico TPS usando dois diferentes plastificantes: glicerol e óleo de buriti, sendo esse óleo um plastificante natural e original para o amido. As blendas PS/TPS foram preparadas em diversas composições por casting e em seguida caracterizadas por termogravimetria (TG, calorimetria exploratória diferencial (DSC, análise termomecânica (TMA e ressonância magnética nuclear (RMN 13C. As blendas PS/TPS com óleo de buriti apresentaram uma contínua redução em suas Tg e Ta. Nas blendas com glicerol esses parâmetros permaneceram praticamente constantes. As blendas também foram submetidas a testes de degradação no solo e depois analisadas por TG e RMN. A análise das blendas PS/TPS com óleo de buriti mostrou que houve um aumento da perda de massa nas curvas TG. Nos espectros de RMN 13C, todos os picos referentes ao amido desapareceram, depois do teste de aterro, sugerindo que houve consumo do amido das blendas por microorganismos. De acordo com as análises realizadas, a adição do TPS produzido com óleo de buriti pode melhorar a degradabilidade do PS.Polystyrene (PS is one of the most used disposable materials in our daily life, but it does not decompose easily. The addition of cassava starch to PS can improve its degradability since starch is a biodegradable polymer. The aim of this work was to develop blends of PS and thermoplastic starch (TPS using two different plasticizers: buriti glycerol and buriti oil, a novel and natural plasticizer for starch. PS/TPS blends were prepared in various contents with the casting technique and analyzed by thermogravimetry (TG, differential scanning calorimetry (DSC

  3. Comparative Characterization of Total Flavonol Glycosides and Terpene Lactones at Different Ages, from Different Cultivation Sources and Genders of Ginkgo biloba Leaves

    Directory of Open Access Journals (Sweden)

    Yong Qin

    2012-08-01

    Full Text Available The extract from Ginkgo biloba leaves has become a very popular plant medicine and herbal supplement for its potential benefit in alleviating symptoms associated with peripheral vascular disease, dementia, asthma and tinnitus. Most research on G. biloba leaves focus on the leaves collected in July and August from four to seven year-old trees, however a large number of leaves from fruit cultivars (trees older than 10 years are ignored and become obsolete after fruit harvest season (November. In this paper, we expand the tree age range (from one to 300 years and first comparatively analyze the total flavonol glycosides and terpene lactones at different ages, from different cultivation sources and genders of G. biloba leaves collected in November by using the validated HPLC-ELSD and HPLC-PDA methods. The results show that the contents of total terpene lactones and flavonol glycosides in the leaves of young ginkgo trees are higher than those in old trees, and they are higher in male trees than in female trees. Geographical factors appear to have a significant influence on the contents as well. These results will provide a good basis for the comprehensive utilization of G. biloba leaves, especially the leaves from fruit cultivars.

  4. Differential response of terpenes and anthraquinones derivatives in Rumex dentatus and Lavandula officinalis to harsh winters across north-western Himalaya.

    Science.gov (United States)

    Jan, Sumira; Kamili, Azra N; Parray, Javid A; Bedi, Yashbir S

    2016-01-01

    Herbs adapted to diverse climates exhibit distinct variability to fluctuating temperatures and demonstrate various metabolic and physiological adaptations to harsh environments. In this research, Rumex dentatus L. and Lavandula officinalis L. were collected before snowfall in September-November to evaluate variability in major phytoconstituents to diverse seasonal regime. LC-MS was used for simultaneous determination of eight anthraquinone derivatives in R. dentatus, i.e. emodin, physcion, chrysophanol, physcion glucoside, endocrocin, emodin glucoside, chrysophanol glucoside and chromone derivatives and monoterpenes in L. officinalis i.e. (Z)-β-ocimene, (E)-β-ocimene, terpene alcohol, terpin-4-ol, acetate ester-linalyl acetate and bicyclic sesquiterpene (E)-caryophyllene. The correlation analysis confirmed significant variation in anthraquinone glucoside and terpene content within Rumex and Lavender, respectively, and altitude was established as the determinant factor in secondary metabolism of both herbs. The study concludes the propagation of herbs in bioclimatic belts which favour accumulation of major constituents and validate their greater pharmacological activity.

  5. Discovery of germacrene A synthases in Barnadesia spinosa: The first committed step in sesquiterpene lactone biosynthesis in the basal member of the Asteraceae

    International Nuclear Information System (INIS)

    Nguyen, Trinh-Don; Faraldos, Juan A.; Vardakou, Maria; Salmon, Melissa; O'Maille, Paul E.; Ro, Dae-Kyun

    2016-01-01

    The Andes-endemic Barnadesioideae lineage is the oldest surviving and phylogenetically basal subfamily of the Asteraceae (Compositae), a prolific group of flowering plants with world-wide distribution (∼24,000 species) marked by a rich diversity of sesquiterpene lactones (STLs). Intriguingly, there is no evidence that members of the Barnadesioideae produce STLs, specialized metabolites thought to have contributed to the adaptive success of the Asteraceae family outside South America. The biosynthesis of STLs requires the intimate expression and functional integration of germacrene A synthase (GAS) and germacrene A oxidase (GAO) to sequentially cyclize and oxidize farnesyl diphosphate into the advanced intermediate germacrene A acid leading to diverse STLs. Our previous discovery of GAO activity conserved across all major subfamilies of Asteraceae, including the phylogenetically basal lineage of Barnadesioideae, prompted further investigation of the presence of the gateway GAS in Barnadesioideae. Herein we isolated two terpene synthases (BsGAS1/BsGAS2) from the basal Barnadesia spinosa (Barnadesioideae) that displayed robust GAS activity when reconstituted in yeast and characterized in vitro. Despite the apparent lack of STLs in the Barnadesioideae, this work unambiguously confirms the presence of GAS in the basal genera of the Asteraceae. Phylogenetic analysis reveals that the two BsGASs fall into two distinct clades of the Asteraceae's GASs, and BsGAS1 clade is only retained in the evolutionary closer Cichorioideae subfamily, implicating BsGAS2 is likely the ancestral base of most GASs found in the lineages outside the Barnadesioideae. Taken together, these results show the enzymatic capacities of GAS and GAO emerged prior to the subsequent radiation of STL-producing Asteraceae subfamilies. - Highlights: • Sesquiterpene lactones are characteristic metabolites in Asteraceae family. • Barnadesioideae is the basal lineage of all Asteraceae plants

  6. Terpene metabolic engineering via nuclear or chloroplast genomes profoundly and globally impacts off-target pathways through metabolite signalling.

    Science.gov (United States)

    Pasoreck, Elise K; Su, Jin; Silverman, Ian M; Gosai, Sager J; Gregory, Brian D; Yuan, Joshua S; Daniell, Henry

    2016-09-01

    The impact of metabolic engineering on nontarget pathways and outcomes of metabolic engineering from different genomes are poorly understood questions. Therefore, squalene biosynthesis genes FARNESYL DIPHOSPHATE SYNTHASE (FPS) and SQUALENE SYNTHASE (SQS) were engineered via the Nicotiana tabacum chloroplast (C), nuclear (N) or both (CN) genomes to promote squalene biosynthesis. SQS levels were ~4300-fold higher in C and CN lines than in N, but all accumulated ~150-fold higher squalene due to substrate or storage limitations. Abnormal leaf and flower phenotypes, including lower pollen production and reduced fertility, were observed regardless of the compartment or level of transgene expression. Substantial changes in metabolomes of all lines were observed: levels of 65-120 unrelated metabolites, including the toxic alkaloid nicotine, changed by as much as 32-fold. Profound effects of transgenesis on nontarget gene expression included changes in the abundance of 19 076 transcripts by up to 2000-fold in CN; 7784 transcripts by up to 1400-fold in N; and 5224 transcripts by as much as 2200-fold in C. Transporter-related transcripts were induced, and cell cycle-associated transcripts were disproportionally repressed in all three lines. Transcriptome changes were validated by qRT-PCR. The mechanism underlying these large changes likely involves metabolite-mediated anterograde and/or retrograde signalling irrespective of the level of transgene expression or end product, due to imbalance of metabolic pools, offering new insight into both anticipated and unanticipated consequences of metabolic engineering. © 2016 The Authors. Plant Biotechnology Journal published by Society for Experimental Biology and The Association of Applied Biologists and John Wiley & Sons Ltd.

  7. Highly divergent mitochondrial ATP synthase complexes in Tetrahymena thermophila.

    Directory of Open Access Journals (Sweden)

    Praveen Balabaskaran Nina

    2010-07-01

    Full Text Available The F-type ATP synthase complex is a rotary nano-motor driven by proton motive force to synthesize ATP. Its F(1 sector catalyzes ATP synthesis, whereas the F(o sector conducts the protons and provides a stator for the rotary action of the complex. Components of both F(1 and F(o sectors are highly conserved across prokaryotes and eukaryotes. Therefore, it was a surprise that genes encoding the a and b subunits as well as other components of the F(o sector were undetectable in the sequenced genomes of a variety of apicomplexan parasites. While the parasitic existence of these organisms could explain the apparent incomplete nature of ATP synthase in Apicomplexa, genes for these essential components were absent even in Tetrahymena thermophila, a free-living ciliate belonging to a sister clade of Apicomplexa, which demonstrates robust oxidative phosphorylation. This observation raises the possibility that the entire clade of Alveolata may have invented novel means to operate ATP synthase complexes. To assess this remarkable possibility, we have carried out an investigation of the ATP synthase from T. thermophila. Blue native polyacrylamide gel electrophoresis (BN-PAGE revealed the ATP synthase to be present as a large complex. Structural study based on single particle electron microscopy analysis suggested the complex to be a dimer with several unique structures including an unusually large domain on the intermembrane side of the ATP synthase and novel domains flanking the c subunit rings. The two monomers were in a parallel configuration rather than the angled configuration previously observed in other organisms. Proteomic analyses of well-resolved ATP synthase complexes from 2-D BN/BN-PAGE identified orthologs of seven canonical ATP synthase subunits, and at least 13 novel proteins that constitute subunits apparently limited to the ciliate lineage. A mitochondrially encoded protein, Ymf66, with predicted eight transmembrane domains could be a

  8. Sucrose Phosphate Synthase and Sucrose Accumulation at Low Temperature 1

    Science.gov (United States)

    Guy, Charles L.; Huber, Joan L. A.; Huber, Steven C.

    1992-01-01

    The influence of growth temperature on the free sugar and sucrose phosphate synthase content and activity of spinach (Spinacia oleracea) leaf tissue was studied. When plants were grown at 25°C for 3 weeks and then transferred to a constant 5°C, sucrose, glucose, and fructose accumulated to high levels during a 14-d period. Predawn sugar levels increased from 14- to 20-fold over the levels present at the outset of the low-temperature treatment. Sucrose was the most abundant free sugar before, during, and after exposure to 5°C. Leaf sucrose phosphate synthase activity was significantly increased by the low-temperature treatment, whereas sucrose synthase and invertases were not. Synthesis of the sucrose phosphate synthase subunit was increased during and after low-temperature exposure and paralleled an increase in the steady-state level of the subunit. The increases in sucrose and its primary biosynthetic enzyme, sucrose phosphate synthase, are discussed in relation to adjustment of metabolism to low nonfreezing temperature and freezing stress tolerance. Images Figure 1 Figure 2 Figure 3 PMID:16652990

  9. Unified Singularity Modeling and Reconfiguration of 3rTPS Metamorphic Parallel Mechanisms with Parallel Constraint Screws

    Directory of Open Access Journals (Sweden)

    Yufeng Zhuang

    2015-01-01

    Full Text Available This paper presents a unified singularity modeling and reconfiguration analysis of variable topologies of a class of metamorphic parallel mechanisms with parallel constraint screws. The new parallel mechanisms consist of three reconfigurable rTPS limbs that have two working phases stemming from the reconfigurable Hooke (rT joint. While one phase has full mobility, the other supplies a constraint force to the platform. Based on these, the platform constraint screw systems show that the new metamorphic parallel mechanisms have four topologies by altering the limb phases with mobility change among 1R2T (one rotation with two translations, 2R2T, and 3R2T and mobility 6. Geometric conditions of the mechanism design are investigated with some special topologies illustrated considering the limb arrangement. Following this and the actuation scheme analysis, a unified Jacobian matrix is formed using screw theory to include the change between geometric constraints and actuation constraints in the topology reconfiguration. Various singular configurations are identified by analyzing screw dependency in the Jacobian matrix. The work in this paper provides basis for singularity-free workspace analysis and optimal design of the class of metamorphic parallel mechanisms with parallel constraint screws which shows simple geometric constraints with potential simple kinematics and dynamics properties.

  10. Analysis of effective thermal conductivity for mineral cast material structures with varying epoxy content using TPS method

    Directory of Open Access Journals (Sweden)

    A. Selvakumar

    2012-01-01

    Full Text Available Conventionally, cast iron is the material used for high speed machine tool structures. As an alternate material to improve the structural properties, composite materials are being used, which are known to exhibit excellent thermal and mechanical properties. While selecting an alternate material, thermal conductivity is an important thermo physical property of the material that should be studied. A resin composite material has a lesser thermal conductivity and its thermal properties vary with the composition of the mixture. A material with lower thermal conductivity will have higher heat concentration within the structure, which may result in structural deformation. In this analysis, epoxy granite, a material which is tested to exhibit excellent mechanical properties has been selected to study its thermal properties. Tests are carried out using Transient Plane Source (TPS method, on eight samples with varying volume fraction of epoxy content in the range 10-24%. It is observed that, the effective thermal conductivity decreases with an increase in epoxy resin content in the mixture because the resin content increases interfacial resistance between particles. Hence, lower epoxy content in the mixture that maximizes the effective thermal conductivity while maintaining good mechanical properties is to be selected.

  11. Analysis of effective thermal conductivity for mineral cast material structures with varying epoxy content using TPS method

    Directory of Open Access Journals (Sweden)

    A. Selvakumar

    2013-04-01

    Full Text Available Conventionally, cast iron is the material used for high speed machine tool structures. As an alternate material to improve the structural properties, composite materials are being used, which are known to exhibit excellent thermal and mechanical properties. While selecting an alternate material, thermal conductivity is an important thermo physical property of the material that should be studied. A resin composite material has a lesser thermal conductivity and its thermal properties vary with the composition of the mixture. A material with lower thermal conductivity will have higher heat concentration within the structure, which may result in structural deformation. In this analysis, epoxy granite, a material which is tested to exhibit excellent mechanical properties has been selected to study its thermal properties. Tests are carried out using Transient Plane Source (TPS method, on eight samples with varying volume fraction of epoxy content in the range 10-24%. It is observed that, the effective thermal conductivity decreases with an increase in epoxy resin content in the mixture because the resin content increases interfacial resistance between particles. Hence, lower epoxy content in the mixture that maximizes the effective thermal conductivity while maintaining good mechanical properties is to be selected.

  12. Control and diagnostics of technical state of main electro technical and thermal equipment of NPP, TPS and HPS on the basis of thermovision technologies

    International Nuclear Information System (INIS)

    Banduryan, B.B.; Fedorenko, G.M.; Ostapchuk, L.B.; Saratov, V.O.

    2006-01-01

    The opportunity of using of thermovision technologies for detection of defects NPP, TPS and HPS electro technical and power equipment is shown. The results of thermovision monitoring of a turbine generator stator technical state are represented. The method for thermovision control of a technical state of a electrical machines and apparatus construction elements, for which the patent of Ukraine have been obtained, is described. he verification of the offered expedient thermovision control and diagnostics at 'Elektrovazhmash' factory was carried out

  13. Principles of Carbon Catabolite Repression in the Rice Blast Fungus: Tps1, Nmr1-3, and a MATE–Family Pump Regulate Glucose Metabolism during Infection

    Science.gov (United States)

    Hartline, David; Quispe, Cristian F.; Madayiputhiya, Nandakumar; Wilson, Richard A.

    2012-01-01

    Understanding the genetic pathways that regulate how pathogenic fungi respond to their environment is paramount to developing effective mitigation strategies against disease. Carbon catabolite repression (CCR) is a global regulatory mechanism found in a wide range of microbial organisms that ensures the preferential utilization of glucose over less favourable carbon sources, but little is known about the components of CCR in filamentous fungi. Here we report three new mediators of CCR in the devastating rice blast fungus Magnaporthe oryzae: the sugar sensor Tps1, the Nmr1-3 inhibitor proteins, and the multidrug and toxin extrusion (MATE)–family pump, Mdt1. Using simple plate tests coupled with transcriptional analysis, we show that Tps1, in response to glucose-6-phosphate sensing, triggers CCR via the inactivation of Nmr1-3. In addition, by dissecting the CCR pathway using Agrobacterium tumefaciens-mediated mutagenesis, we also show that Mdt1 is an additional and previously unknown regulator of glucose metabolism. Mdt1 regulates glucose assimilation downstream of Tps1 and is necessary for nutrient utilization, sporulation, and pathogenicity. This is the first functional characterization of a MATE–family protein in filamentous fungi and the first description of a MATE protein in genetic regulation or plant pathogenicity. Perturbing CCR in Δtps1 and MDT1 disruption strains thus results in physiological defects that impact pathogenesis, possibly through the early expression of cell wall–degrading enzymes. Taken together, the importance of discovering three new regulators of carbon metabolism lies in understanding how M. oryzae and other pathogenic fungi respond to nutrient availability and control development during infection. PMID:22570632

  14. EKSPERIMENTASI MODEL PEMBELAJARAN KOOPERATIF TIPE THINK PAIR SHARE (TPS DENGAN PENDEKATAN OPEN-ENDED DITINJAU DARI DISPOSISI MATEMATIS SISWA KELAS VIII SMP NEGERI DI KABUPATEN PURWOREJO

    Directory of Open Access Journals (Sweden)

    Zainnur Wijayanto

    2015-02-01

    Full Text Available prestasi belajar matematika lebih baik, siswa dengan disposisi matematis tinggi, sedang, atau rendah, 4 pada masing-masing kategori disposisi matematis, manakah yang mempunyai prestasi belajar matematika lebih baik, siswa yang dikenai model pembelajaran kooperatif tipe TPS dengan pendekatan open-ended atau pembelajaran langsung. Penelitian ini merupakan penelitian eksperimental semu. Populasi penelitian ini adalah siswa kelas VIII SMP Negeri di Kabupaten Purworejo tahun ajaran 2013/2014. Sampel dalam penelitian ini adalah siswa SMPN 20 Purworejo, SMPN 14 Purworejo, SMPN 13 Purworejo yang diambil secara stratified cluster random sampling. Instrumen yang digunakan untuk mengumpulkan data adalah instrumen tes prestasi belajar matematika, instrumen angket disposisi matematis siswa. Teknik analisis data yang digunakan adalah uji hipotesis menggunakan anava dua jalan dengan sel tak sama. Dari hasil analisis disimpulkan bahwa : 1 prestasi belajar matematika siswa yang diberi pembelajaran dengan model pembelajaran kooperatif tipe TPS dengan pendekatan open-ended lebih baik daripada siswa yang diberi pembelajaran langsung, 2 prestasi belajar matematika siswa dengan disposisi matematis tinggi lebih baik daripada siswa dengan disposisi matematis sedang dan rendah, begitu juga dengan prestasi belajar matematika siswa dengan disposisi matematis sedang lebih baik daripada siswa dengan disposisi matematis rendah, 3 pada model pembelajaran kooperatif tipe TPS dengan pendekatan open-ended prestasi belajar matematika siswa yang memiliki disposisi matematis tinggi lebih baik daripada siswa yang memiliki disposisi matematis sedang maupun rendah. Pada pembelajaran langsung, siswa dengan disposisi matematis tinggi memiliki prestasi belajar yang lebih baik daripada siswa yang memiliki disposisi matematis sedang dan rendah, siswa dengan disposisi matematis sedang memiliki prestasi belajar yang sama dengan siswa yang memiliki disposisi matematis rendah, 4 Pada siswa

  15. EKSPERIMENTASI MODEL PEMBELAJARAN KOOPERATIF TIPE THINK PAIR SHARE (TPS DITINJAU DARI KEMANDIRIAN BELAJAR SISWA KELAS VIII SMP NEGERI DI KOTA YOGYAKARTA

    Directory of Open Access Journals (Sweden)

    Annis Deshinta Ayuningtyas

    2015-02-01

    Full Text Available Tujuan penelitian ini yaitu untuk mengetahui: (1 manakah yang memberikan prestasi belajar lebih baik diantara model pembelajaran kooperatif tipe TPSatau pembelajaran langsung, (2 manakah yang mempunyai prestasi belajar lebih baik di antara siswa yang mempunyai kemandirian belajar tinggi, sedang, atau rendah, (3 pada masing-masing tingkatan kemandirian belajar (tinggi, sedang, dan rendah, manakah yang memberikan prestasi belajar lebih baik antara model pembelajaran kooperatif tipe TPS atau pembelajaran langsung. Penelitian ini termasuk penelitian eksperimental semu dengan desain factorial 2x3. Populasi dalam penelitian ini adalah seluruh siswa kelas VIII Sekolah Menengah Pertama (SMP Negeri Se-Kota Yogyakarta Semester I tahun pelajaran 2011/2012. Teknik pengambilan sampel dilakukan dengan carastratified cluster random sampling. Banyak sampel adalah 195 siswa. Instrumen yang digunakan untuk mengumpulkan data adalah angket kemandirian belajar siswa dan tes prestasi belajar matematika. Uji coba angket meliputi validitas isi, konsistensi internal dan reliabilitas. Uji coba tes prestasi meliputi validitas isi, tingkat kesukaran, daya beda dan reliabilitas. Untuk uji normalitas menggunakan uji Lilliefors, uji homogenitas menggunakan uji barlett. Uji keseimbangan yang digunakan yaitu anava satu jalan dengan sel tak sama. Analisis tes prestasi matematika menggunakan analisis variansi dua jalan dengan sel tak sama. Dari hasil analisis diperoleh kesimpulan bahwa: (1 Prestasi belajar matematika dengan model pembelajaran kooperatiftipe TPS memberikan prestasi belajar yang lebih baik daripada model pembelajaran langsung, (2 Siswa yang kemandirian belajarnya tinggi mempunyai prestasi belajar yang lebih baik daripada siswa dengan kemandirian belajar sedang dan rendah, sedangkan siswa yang kemandirian belajarnya sedang mempunyai prestasi belajar yang sama dengan siswa dengan kemandirian belajar rendah, (3 Pada siswa yang mempunyai kemandirian belajar tinggi

  16. EFEKTIVITAS MODEL PEMBELAJARAN PORTOFOLIO DAN MODEL PEMBELAJARAN KOOPERATIF THINK PAIR AND SHARE (TPS TERHADAP PRESTASI BELAJAR MATEMATIKA DITINJAU DARI KREATIVITAS SISWA VII SMP NEGERI 2 KEBONSARI TAHUN AJARAN 2011/2012

    Directory of Open Access Journals (Sweden)

    Vera Dewi Susanti

    2014-02-01

    Dari hasil analisis data didapatkan: 1 ada perbedaan pengaruh antara model pembelajaran portofolio dan model pembelajaran kooperatif think pair and share (TPS dalam meningkatkan prestasi belajar matematika siswa (nilai signifikansi 0,05.

  17. PERBEDAAN HASIL BELAJAR EKONOMI SISWA YANG MENGGUNAKAN MODEL PEMBELAJARANKOOPERATIFTIPETHINK PAIR SHARE (TPS DENGAN TIPE STUDENT TEAMS ACHIEVEMENT DIVISION(STADSISWA KELAS XI IPS SMAN 11 PADANG

    Directory of Open Access Journals (Sweden)

    Dina Sofianti

    2013-04-01

    Full Text Available The research aims to analyze the difference of learning points by using the cooperative learning models between Think Pair Share (TPS and Student Teams Achievement Division (STAD types of the XI-grade students of IPS SMAN 11 Padang. It is an experimental research. The population of this research is all XI-grade students of IPS SMAN 11 Padang, who were registered in the year 2013-2014. Two samples were selected by using the technique of Purposive Sampling method. The data of this research is primary data, and the data analyses are descriptive and inductive, conducted through t-test by first making tests of normality and homogeneity of both sample variants. Based on the data analyses, it is obtained that the learning points for the experimental class 1 is 76.80 and the experimental class 2 is 65.64 on the average, meaning that Sig 0.018 is smaller than α = 0.05. From the hypothesis tests, tcalc.is 2.444 and ttable is 1.67, so that H0 is rejected, meaning that there is the difference between TPS and STAD. From the results, it is expected that this could provide a solution for the problem of lowering learning points especially in economics subject. Finally, for improving the learning points of economical students, it is suggested for teachers to consider the learning models from Think Pair Share TPS and STAD.

  18. Permanent Seed Implant Dosimetry (PSID)TM 4.5 version as isodose and Treatment Planning System (TPS) programme for brachytherapy

    International Nuclear Information System (INIS)

    Indra Saptiama; Moch Subechi; Anung Pujiyanto; Hotman Lubis; Herlan Setiawan

    2014-01-01

    The medical treatment using radiation therapy for cancer diseases is increasingly developed. One of the method used in radiotherapy is brachytherapy. Brachytherapy is radiation therapy method in which a radiation source is implanted in cancer cell directly so the dose accepted by cancer cell is the highest dose and the dose accepted by normal cell is the lowest dose. I-125 Seed have been made successfully in domestic. To support the implant of I-125 seed for brachytherapy needs computer programme for the isodose calculation and Treatment Planning System (TPS). Permanent Seed Implant Dosimetry (PSID) 4.5 is one of the isodose calculation and Treatment Planning System (TPS) programme that is owned by Center for Radioisotope and Radiopharmaceutical-BATAN. In isodose calculation, PSID 4.5 uses 1D formalism and 2D formalism based on AAPM-TG43 (Association of American Physicist in Medicine- Task Group No.43). Anisotropic function on 1D formalism depend on distance function while on 2D formalism count on distance and angle function therefore 2D formalism has isodose calculation better than 1D formalism usage. PSID 4.5 can display the isodose contour of the seed I-125 radiation source in 2 dimension (2D) and 3 dimension (3D). The computer programme of isodose calculation and TPS uses PSID 4.5 is expected able to help planning for seed I-125 implantation process for brachytherapy that used by paramedics and to support the usage of seed I-125 as domestic product. (author)

  19. Arabidopsis CDS blastp result: AK241679 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK241679 J065193F24 At3g29410.1 68416.m03695 terpene synthase/cyclase family protein similar to terpene... synthase GB:CAA72074 from [Arabidopsis thaliana], contains Pfam profile: PF01397 terpene synthase family 5e-65 ...

  20. Arabidopsis CDS blastp result: AK242212 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242212 J075171E13 At3g29410.1 68416.m03695 terpene synthase/cyclase family protein similar to terpene... synthase GB:CAA72074 from [Arabidopsis thaliana], contains Pfam profile: PF01397 terpene synthase family 1e-21 ...

  1. Impaired glycogen synthase activity and mitochondrial dysfunction in skeletal muscle

    DEFF Research Database (Denmark)

    Højlund, Kurt; Beck-Nielsen, Henning

    2006-01-01

    Insulin resistance in skeletal muscle is a major hallmark of type 2 diabetes and an early detectable abnormality in the development of this disease. The cellular mechanisms of insulin resistance include impaired insulin-mediated muscle glycogen synthesis and increased intramyocellular lipid content......, whereas impaired insulin activation of muscle glycogen synthase represents a consistent, molecular defect found in both type 2 diabetic and high-risk individuals. Despite several studies of the insulin signaling pathway believed to mediate dephosphorylation and hence activation of glycogen synthase......, the molecular mechanisms responsible for this defect remain unknown. Recently, the use of phospho-specific antibodies in human diabetic muscle has revealed hyperphosphorylation of glycogen synthase at sites not regulated by the classical insulin signaling pathway. In addition, novel approaches such as gene...

  2. Class II recombinant phosphoribosyl diphosphate synthase from spinach

    DEFF Research Database (Denmark)

    Krath, B N; Hove-Jensen, B

    2001-01-01

    to other PRPP synthases the activity of spinach PRPP synthase isozyme 3 is independent of P(i), and the enzyme is inhibited by ribonucleoside diphosphates in a purely competitive manner, which indicates a lack of allosteric inhibition by these compounds. In addition spinach PRPP synthase isozyme 3 shows...... an unusual low specificity toward diphosphoryl donors by accepting dATP, GTP, CTP, and UTP in addition to ATP. The kinetic mechanism of the enzyme is an ordered steady state Bi Bi mechanism with K(ATP) and K(Rib-5-P) values of 170 and 110 micrometer, respectively, and a V(max) value of 13.1 micromol (min x...... mg of protein)(-1). The enzyme has an absolute requirement for magnesium ions, and maximal activity is obtained at 40 degrees C at pH 7.6....

  3. The diagnostic value of serum tumor markers CEA, CA19-9, CA125, CA15-3, and TPS in metastatic breast cancer.

    Science.gov (United States)

    Wang, Weigang; Xu, Xiaoqin; Tian, Baoguo; Wang, Yan; Du, Lili; Sun, Ting; Shi, Yanchun; Zhao, Xianwen; Jing, Jiexian

    2017-07-01

    This study aims to understand the diagnostic value of serum tumor markers carcinoembryonic antigen (CEA), cancer antigen 19-9 (CA19-9), cancer antigen 125 (CA125), cancer antigen 15-3 (CA15-3), and tissue polypeptide-specific antigen (TPS) in metastatic breast cancer (MBC). A total of 164 metastatic breast cancer patients in Shanxi Cancer Hospital were recruited between February 2016 and July 2016. 200 breast cancer patients without metastasis in the same period were randomly selected as the control group. The general characteristics, immunohistochemical, and pathological results were investigated between the two groups, and tumor markers were determined. There were statistical differences in the concentration and the positive rates of CEA, CA19-9, CA125, CA15-3, and TPS between the MBC and control group (Ptumor marker at 56.7% and 97.0%, respectively. In addition, two tumor markers were used for the diagnosis of MBC and the CEA and TPS combination had the highest diagnostic sensitivity with 78.7%, while the CA15-3 and CA125 combination had the highest specificity of 91.5%. Analysis of tumor markers of 164 MBC found that there were statistical differences in the positive rates of CEA and CA15-3 between bone metastases and other metastases (χ 2 =6.00, P=0.014; χ 2 =7.32, P=0.007, respectively). The sensitivity and specificity values of the CEA and CA15-3 combination in the diagnosis of bone metastases were 77.1% and 45.8%, respectively. The positive rate of TPS in the lung metastases group was lower than in other metastases (χ 2 =8.06, P=0.005).There were significant differences in the positive rates of CA15-3 and TPS between liver metastases and other metastases (χ 2 =15.42, Ptumor markers have varying diagnostic value. Copyright © 2017 Elsevier B.V. All rights reserved.

  4. Virtual Screening of Novel Glucosamine-6-Phosphate Synthase Inhibitors.

    Science.gov (United States)

    Lather, Amit; Sharma, Sunil; Khatkar, Anurag

    2018-01-01

    Infections caused by microorganisms are the major cause of death today. The tremendous and improper use of antimicrobial agents leads to antimicrobial resistance. Various currently available antimicrobial drugs are inadequate to control the infections and lead to various adverse drug reactions. Efforts based on computer-aided drug design (CADD) can excavate a large number of databases to generate new, potent hits and minimize the requirement of time as well as money for the discovery of newer antimicrobials. Pharmaceutical sciences also have made development with advances in drug designing concepts. The current research article focuses on the study of various G-6-P synthase inhibitors from literature cited molecular database. Docking analysis was conducted and ADMET data of various molecules was evaluated by Schrodinger Glide and PreADMET software, respectively. Here, the results presented efficacy of various inhibitors towards enzyme G-6-P synthase. Docking scores, binding energy and ADMET data of various molecules showed good inhibitory potential toward G-6-P synthase as compared to standard antibiotics. This novel antimicrobial drug target G-6-P synthase has not so extensively been explored for its application in antimicrobial therapy, so the work done so far proved highly essential. This article has helped the drug researchers and scientists to intensively explore about this wonderful antimicrobial drug target. The Schrodinger, Inc. (New York, USA) software was utilized to carry out the computational calculations and docking studies. The hardware configuration was Intel® core (TM) i5-4210U CPU @ 2.40GHz, RAM memory 4.0 GB under 64-bit window operating system. The ADMET data was calculated by using the PreADMET tool (PreADMET ver. 2.0). All the computational work was completed in the Laboratory for Enzyme Inhibition Studies, Department of Pharmaceutical Sciences, M.D. University, Rohtak, INDIA. Molecular docking studies were carried out to identify the binding

  5. Structure and mechanism of the diterpene cyclase ent-copalyl diphosphate synthase

    Energy Technology Data Exchange (ETDEWEB)

    Köksal, Mustafa; Hu, Huayou; Coates, Robert M.; Peters, Reuben J.; Christianson, David W. (UIUC); (Iowa State); (Penn)

    2011-09-20

    The structure of ent-copalyl diphosphate synthase reveals three {alpha}-helical domains ({alpha}, {beta} and {gamma}), as also observed in the related diterpene cyclase taxadiene synthase. However, active sites are located at the interface of the {beta}{gamma} domains in ent-copalyl diphosphate synthase but exclusively in the {alpha} domain of taxadiene synthase. Modular domain architecture in plant diterpene cyclases enables the evolution of alternative active sites and chemical strategies for catalyzing isoprenoid cyclization reactions.

  6. Highly Divergent Mitochondrial ATP Synthase Complexes in Tetrahymena thermophila

    NARCIS (Netherlands)

    Nina, Praveen Balabaskaran; Dudkina, Natalya V.; Kane, Lesley A.; van Eyk, Jennifer E.; Boekema, Egbert J.; Mather, Michael W.; Vaidya, Akhil B.; Eisen, Jonathan A.

    The F-type ATP synthase complex is a rotary nano-motor driven by proton motive force to synthesize ATP. Its F(1) sector catalyzes ATP synthesis, whereas the F(o) sector conducts the protons and provides a stator for the rotary action of the complex. Components of both F(1) and F(o) sectors are

  7. Predicting the catalytic sites of isopenicillin N synthase (IPNS ...

    African Journals Online (AJOL)

    Isopenicillin N synthase (IPNS) related Non-haem iron-dependent oxygenases and oxidases (NHIDOX) demonstrated a striking structural conservativeness, even with low protein sequence homology. It is evident that these enzymes have an architecturally similar catalytic centre with active ligands lining the reactive pocket.

  8. Studies on the Active Site of Deacetoxycephalosporin C Synthase

    NARCIS (Netherlands)

    Lloyd, Matthew D.; Lee, Hwei-Jen; Harlos, Karl; Zhang, Zhi-Hong; Baldwin, Jack E.; Schofield, Christopher J.; Charnock, John M.; Garner, C. David; Hara, Takane; Terwisscha van Scheltinga, Anke C.; Valegård, Karin; Viklund, Jenny A.C.; Hajdu, Janos; Andersson, Inger; Danielsson, Åke; Bhikhabhai, Rama

    1999-01-01

    The Fe(II) and 2-oxoglutarate-dependent dioxygenase deacetoxycephalosporin C synthase (DAOCS) from Streptomyces clavuligerus was expressed at ca 25% of total soluble protein in Escherichia coli and purified by an efficient large-scale procedure. Purified protein catalysed the conversions of

  9. Beta-Glucan Synthase Gene Expression in Pleurotus sp

    International Nuclear Information System (INIS)

    Azhar Mohamad; Nie, H.J.

    2016-01-01

    Pleurotus sp. is a popular edible mushroom, containing various functional component, in particular, Beta-glucan. Beta-glucans is a part of glucan family of polysaccharides and supposedly contribute to medicinal and nutritional value of Pleurotus.sp. In order to understand the distribution of Beta-glucan in Pleurotus.sp, the Beta-glucan synthase gene expression was determined and compared in different part of Pleurotus, namely mycelium, stripe and cap. The Pleurotus.sp RNA was extracted using commercial kit, employing Tissuelyser ll (Qiagen, USA) to disrupt the cell walls. Then the RNA was quantified by Nano drop (Thermo Fisher, USA) and visualized using denaturing agarose gel. RNA with good OD 260.280 reading (∼2.0) was chosen and converted to cDNA. Using Laccase synthase gene as home keeping gene, Beta-glucan synthase gene expression was quantified using CFX 96 Real Time PCR detection system (Biorad, USA). Preliminary result shows that Beta-glucan synthase was relatively expressed the most in stripe, followed by mycelium and barely in cap. (author)

  10. Isolation and expression of the Pneumocystis carinii thymidylate synthase gene

    DEFF Research Database (Denmark)

    Edman, U; Edman, J C; Lundgren, B

    1989-01-01

    The thymidylate synthase (TS) gene from Pneumocystis carinii has been isolated from complementary and genomic DNA libraries and expressed in Escherichia coli. The coding sequence of TS is 891 nucleotides, encoding a 297-amino acid protein of Mr 34,269. The deduced amino acid sequence is similar...

  11. Endothelial nitric oxide synthase polymorphism G298T in ...

    Indian Academy of Sciences (India)

    Supplementary data: Endothelial nitric oxide synthase polymorphism G298T in association with oxidative DNA damage in coronary atherosclerosis. Rajesh G. Kumar, Mrudula K. Spurthi, Kishore G. Kumar, Sanjib K. Sahu and Surekha H. Rani. J. Genet. 91, 349–352. Table 1. The demographic and clinical data of the CHD ...

  12. ATP synthase--a marvellous rotary engine of the cell.

    Science.gov (United States)

    Yoshida, M; Muneyuki, E; Hisabori, T

    2001-09-01

    ATP synthase can be thought of as a complex of two motors--the ATP-driven F1 motor and the proton-driven Fo motor--that rotate in opposite directions. The mechanisms by which rotation and catalysis are coupled in the working enzyme are now being unravelled on a molecular scale.

  13. Contribution of granule bound starch synthase in kernel modification ...

    African Journals Online (AJOL)

    The role of gbssI and gbssII genes, encoding granule bound starch synthase enzyme I and II, respectively, in quality protein maize (QPM) were studied at different days after pollination (DAP). Total RNA was used for first strand cDNA synthesis using the ImpromIISriptTM reverse transcriptase. No detectable levels of gbssI ...

  14. Dihydropteroate synthase gene mutations in Pneumocystis and sulfa resistance

    DEFF Research Database (Denmark)

    Huang, Laurence; Crothers, Kristina; Atzori, Chiara

    2004-01-01

    in the dihydropteroate synthase (DHPS) gene. Similar mutations have been observed in P. jirovecii. Studies have consistently demonstrated a significant association between the use of sulfa drugs for PCP prophylaxis and DHPS gene mutations. Whether these mutations confer resistance to TMP-SMX or dapsone plus trimethoprim...

  15. Analysis of genetic variation of inducible nitric oxide synthase and ...

    African Journals Online (AJOL)

    The genetic diversity of 100 Malaysian native chickens was investigated using polymerase chain reaction-restriction fragment polymorphism (PCR-RFLP) for two candidate genes: inducible nitric oxide synthase (INOS) and natural resistance-associated macrophage protein 1 (NRAMP1). The two genes were selected ...

  16. Characterising the cellulose synthase complexes of cell walls

    NARCIS (Netherlands)

    Mansoori Zangir, N.

    2012-01-01

    One of the characteristics of the plant kingdom is the presence of a structural cell wall. Cellulose is a major component in both the primary and secondary cell walls of plants. In higher plants cellulose is synthesized by so called rosette protein complexes with cellulose synthases (CESAs) as

  17. Molecular cloning and characterization of strictosidine synthase, a ...

    African Journals Online (AJOL)

    Mitragynine is one of the most dominant indole alkaloids present in the leaves of Mitragyna speciosa, a species of Rubiaceae. This alkaloid is believed to be synthesized via condensation of the amino acid derivative, tryptamine and secologanine by the action of strictosidine synthase (STR). The cDNA clone encoding STR ...

  18. Alternating copolymerization of propylene oxide with biorenewable terpene-based cyclic anhydrides: a sustainable route to aliphatic polyesters with high glass transition temperatures.

    Science.gov (United States)

    Van Zee, Nathan J; Coates, Geoffrey W

    2015-02-23

    The alternating copolymerization of propylene oxide with terpene-based cyclic anhydrides catalyzed by chromium, cobalt, and aluminum salen complexes is reported. The use of the Diels-Alder adduct of α-terpinene and maleic anhydride as the cyclic anhydride comonomer results in amorphous polyesters that exhibit glass transition temperatures (Tg ) of up to 109 °C. The polymerization conditions and choice of catalyst have a dramatic impact on the molecular weight distribution, the relative stereochemistry of the diester units along the polymer chain, and ultimately the Tg of the resulting polymer. The aluminum salen complex exhibits exceptional selectivity for copolymerization without transesterification or epimerization side reactions. The resulting polyesters are highly alternating and have high molecular weights and narrow polydispersities. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  19. Seasonal Terpene Variation in Needles of Pinus radiata (Pinales: Pinaceae) Trees Attacked by Tomicus piniperda (Coleoptera: Scolytinae) and the Effect of Limonene on Beetle Aggregation.

    Science.gov (United States)

    Romón, Pedro; Aparicio, Domitila; Palacios, Francisco; Iturrondobeitia, Juan Carlos; Hance, Thierry; Goldarazena, Arturo

    2017-09-01

    Concentrations of four monoterpenes were determined in needles of Pinus radiata (D.Don) (Pinales: Pinaceae) trees that were attacked or nonattacked by Tomicus piniperda (L.) (Coleoptera: Scolytinae). Compounds were identified and quantified by gas chromatography-mass spectrometry. The mean ambient temperature was obtained using climate-recording data loggers. The effect of limonene on field aggregation was also evaluated at three limonene release rates using Lindgren attractant-baited traps and trap logs. Attacked trees produced less α-pinene in March, July, and November than nonattacked trees, less β-pinene in July and November, and less limonene from May to November. Limonene reduced the attraction of T. piniperda to attractant-baited traps and trap logs. Results were linked to better responses to high temperatures, with respect to terpene contents, by the nonattacked trees after the spring attack. © The Author 2017. Published by Oxford University Press on behalf of Entomological Society of America.

  20. Valencene synthase from the heartwood of Nootka cypress (Callitropsis nootkatensis) for biotechnological production of valencene

    NARCIS (Netherlands)

    Beekwilder, M.J.; Houwelingen, van A.M.M.L.; Cankar, K.; Dijk, van A.D.J.; Jong, de R.; Stoopen, G.M.; Bouwmeester, H.J.; Achkar, J.; Sonke, Th.; Bosch, H.J.

    2014-01-01

    Nootkatone is one of the major terpenes in the heartwood of the Nootka cypress Callitropsis nootkatensis. It is an oxidized sesquiterpene, which has been postulated to be derived from valencene. Both valencene and nootkatone are used for flavouring citrus beverages and are considered among the most

  1. Identifying the catalytic components of cellulose synthase and the maize mixed-linkage beta-glucan synthase

    Energy Technology Data Exchange (ETDEWEB)

    Nicholas C Carpita

    2009-04-20

    Five specific objectives of this project are to develop strategies to identify the genes that encode the catalytic components of "mixed-linkage" (1→3),(1→4)-beta-D-glucans in grasses, to determine the protein components of the synthase complex, and determine the biochemical mechanism of synthesis. We have used proteomic approaches to define intrinsic and extrinsic polypeptides of Golgi membranes that are associated with polysaccharide synthesis and trafficking. We were successful in producing recombinant catalytic domains of cellulose synthase genes and discovered that they dimerize upon concentration, indicating that two CesA proteins form the catalytic unit. We characterized a brittle stalk2 mutant as a defect in a COBRA-like protein that results in compromised lignin-cellulose interactions that decrease tissue flexibility. We used virus-induced gene silencing of barley cell wall polysaccharide synthesis by BSMV in an attempt to silence specific members of the cellulose synthase-like gene family. However, we unexpectedly found that regardless of the specificity of the target gene, whole gene interaction networks were silenced. We discovered the cause to be an antisense transcript of the cellulose synthase gene initiated small interfering RNAs that spread silencing to related genes.

  2. Evolutionary and mechanistic insights from the reconstruction of α-humulene synthases from a modern (+)-germacrene A synthase.

    Science.gov (United States)

    Gonzalez, Veronica; Touchet, Sabrina; Grundy, Daniel J; Faraldos, Juan A; Allemann, Rudolf K

    2014-10-15

    Germacrene A synthase (GAS) from Solidago canadensis catalyzes the conversion of farnesyl diphosphate (FDP) to the plant sesquiterpene (+)-germacrene A. After diphosphate expulsion, farnesyl cation reacts with the distal 10,11-double bond to afford germacrene A (>96%) and <2% α-humulene, which arises from 1,11-cyclization of FDP. The origin of the 1,11-activity of GAS was investigated by amino acid sequence alignments of 1,10- and 1,11-synthases and comparisons of X-ray crystal structures with the homology model of GAS; a triad [Thr 401-Gly 402-Gly 403] that might be responsible for the predominant 1,10-cyclization activity of GAS was identified. Replacement of Gly 402 with residues of increasing size led to a progressive increase of 1,11-cyclization. The catalytic robustness of these 1,10- /1,11-GAS variants point to Gly 402 as a functional switch of evolutionary significance and suggests that enzymes with strict functionalities have evolved from less specific ancestors through a small number of substitutions. Similar results were obtained with germacrene D synthase (GDS) upon replacement of the homologous active-site residue Gly 404: GDS-G404V generated approximately 20% bicyclogermacrene, a hydrocarbon with a cyclopropane ring that underlines the dual 1,10-/1,11-cyclization activity of this mutant. This suggests that the reaction pathways to germacrenes and humulenes might be connected through a bridged 1,10,11-carbocation intermediate or transition state that resembles bicyclogermacrene. Mechanistic studies using [1-(3)H1]-10-fluorofarnesyl diphosphate and deuterium-labeling experiments with [12,13-(2)H6]-FDP support a germacrene-humulene rearrangement linking 1,10- and 1,11-pathways. These results support the bioinformatics proposal that modern 1,10-synthases could have evolved from promiscuous 1,11-sesquiterpene synthases.

  3. A study to compare the motorised wedge output factor of an elekta synergy linear accelerator with reference data (TPS Data)

    International Nuclear Information System (INIS)

    Akosah, Kinsley

    2016-07-01

    For external beam radiotherapy treatments, high doses are delivered to the cancerous cell. Accuracy and precision of dose delivery are primary requirements for effective and efficient cancer treatment. The dose delivered to the patient might not be uniform and therefore need to be compensated for. In treatment these inhomogeneities are taken care of by using wedge filters and incorporating wedge factors in the Treatment Planning System. Computer controlled wedges were alternatives introduced by different manufacturers of which Motorized wedges (MW) is one of them. The MW was introduced by ELEKTA and this helps to overcome some of the shortcomings of physical wedges. The objectives of this study were to measure MW output factors for 6 MV and 15 MV photon energies for an ELEKTA Synergy. Secondly, to compare the results of MWOF obtained to that of the treatment planning system data. The Motorized Wedge Output Factors (MWOF) were measured for the ELEKTA Synergy for both 6 MV and 15 MV photon energies. With the help of PMMA solid water slabs phantom, the Elekta synergy, thermometer, barometer, PTW farmer type ionization chamber 30010 charges were collected at 100 cm source to surface distance for various square field sizes from 5x5 cm to 30x30 cm and depth of 1.5 cm and 2.5 cm for 6 MV and 15 MV photon energies. Comparing the results with the TPS data, an excellent agreement was found for 6 MV MWOF, with the percentage differences ranging from 0.03% to 1.50%, with a mean of 0.03%. The coefficient of variation of MWOF ranged from 0.023% to 1.07% and 0.001% to 12.89% for the two beams (6 MV and 15 MV) respectively. The relative differences between the calculated and the measured MWOFs increases with field size. In conclusion, there was general agreement between the calculated and measured MWOFs. The consistency of values provide further support that a standard dataset of photon and electron dosimetry could be established as a guide for future commissioning, beam modeling

  4. SU-F-T-47: MRI T2 Exclusive Based Planning Using the Endocavitary/interstitial Gynecological Benidorm Applicator: A Proposed TPS Library and Preplan Efficient Methodology

    Energy Technology Data Exchange (ETDEWEB)

    Richart, J; Otal, A; Rodriguez, S; Santos, M [Clinica Benidorm, Benidorm, Alicante (Spain); Perez-Calatayud, J [Clinica Benidorm, Benidorm, Alicante (Spain); Hospital La Fe, Valencia (Spain)

    2016-06-15

    Purpose: ABS and GEC-ESTRO have recommended MRI T2 for image guided brachytherapy. Recently, a new applicator (Benidorm Template, TB) has been developed in our Department (Rodriguez et al 2015). TB is fully MRI compatible because the Titanium needles and it allows the use of intrauterine tandem. Currently, TPS applicators library are not currently available for non-rigid applicators in case of interstitial component as the TB.The purpose of this work is to present the development of a library for the TB, together with its use on a pre-planning technique. Both new goals allow a very efficient and exclusive T2 MRI based planning clinical TB implementation. Methods: The developed library has been implemented in Oncentra Brachytherapy TPS, version 4.3.0 (Elekta) and now is being implemented on Sagiplan v 2.0 TPS (Eckert&Ziegler BEBIG). To model the TB, free and open software named FreeCAD and MeshLab have been used. The reconstruction process is based on three inserted A-vitamin pellets together with the data provided by the free length. The implemented preplanning procedure is as follow: 1) A MRI T2 acquisition is performed with the template in place just with the vaginal cylinder (no uterine tube nor needles). 2) The CTV is drawn and the required needles are selected using a developed Java based application and 3) A post-implant MRI T2 is performed. Results: This library procedure has been successfully applied by now in 25 patients. In this work the use of the developed library will be illustrated with clinical examples. The preplanning procedure has been applied by now in 6 patients, having significant advantages: needle depth estimation, needle positions and number are optimized a priori, time saving, etc Conclusion: TB library and pre-plan techniques are feasible and very efficient and their use will be illustrated in this work.

  5. SU-F-T-47: MRI T2 Exclusive Based Planning Using the Endocavitary/interstitial Gynecological Benidorm Applicator: A Proposed TPS Library and Preplan Efficient Methodology

    International Nuclear Information System (INIS)

    Richart, J; Otal, A; Rodriguez, S; Santos, M; Perez-Calatayud, J

    2016-01-01

    Purpose: ABS and GEC-ESTRO have recommended MRI T2 for image guided brachytherapy. Recently, a new applicator (Benidorm Template, TB) has been developed in our Department (Rodriguez et al 2015). TB is fully MRI compatible because the Titanium needles and it allows the use of intrauterine tandem. Currently, TPS applicators library are not currently available for non-rigid applicators in case of interstitial component as the TB.The purpose of this work is to present the development of a library for the TB, together with its use on a pre-planning technique. Both new goals allow a very efficient and exclusive T2 MRI based planning clinical TB implementation. Methods: The developed library has been implemented in Oncentra Brachytherapy TPS, version 4.3.0 (Elekta) and now is being implemented on Sagiplan v 2.0 TPS (Eckert&Ziegler BEBIG). To model the TB, free and open software named FreeCAD and MeshLab have been used. The reconstruction process is based on three inserted A-vitamin pellets together with the data provided by the free length. The implemented preplanning procedure is as follow: 1) A MRI T2 acquisition is performed with the template in place just with the vaginal cylinder (no uterine tube nor needles). 2) The CTV is drawn and the required needles are selected using a developed Java based application and 3) A post-implant MRI T2 is performed. Results: This library procedure has been successfully applied by now in 25 patients. In this work the use of the developed library will be illustrated with clinical examples. The preplanning procedure has been applied by now in 6 patients, having significant advantages: needle depth estimation, needle positions and number are optimized a priori, time saving, etc Conclusion: TB library and pre-plan techniques are feasible and very efficient and their use will be illustrated in this work.

  6. SU-E-T-214: Intensity Modulated Proton Therapy (IMPT) Based On Passively Scattered Protons and Multi-Leaf Collimation: Prototype TPS and Dosimetry Study

    International Nuclear Information System (INIS)

    Sanchez-Parcerisa, D; Carabe-Fernandez, A

    2014-01-01

    Purpose. Intensity-modulated proton therapy is usually implemented with multi-field optimization of pencil-beam scanning (PBS) proton fields. However, at the view of the experience with photon-IMRT, proton facilities equipped with double-scattering (DS) delivery and multi-leaf collimation (MLC) could produce highly conformal dose distributions (and possibly eliminate the need for patient-specific compensators) with a clever use of their MLC field shaping, provided that an optimal inverse TPS is developed. Methods. A prototype TPS was developed in MATLAB. The dose calculation process was based on a fluence-dose algorithm on an adaptive divergent grid. A database of dose kernels was precalculated in order to allow for fast variations of the field range and modulation during optimization. The inverse planning process was based on the adaptive simulated annealing approach, with direct aperture optimization of the MLC leaves. A dosimetry study was performed on a phantom formed by three concentrical semicylinders separated by 5 mm, of which the inner-most and outer-most were regarded as organs at risk (OARs), and the middle one as the PTV. We chose a concave target (which is not treatable with conventional DS fields) to show the potential of our technique. The optimizer was configured to minimize the mean dose to the OARs while keeping a good coverage of the target. Results. The plan produced by the prototype TPS achieved a conformity index of 1.34, with the mean doses to the OARs below 78% of the prescribed dose. This Result is hardly achievable with traditional conformal DS technique with compensators, and it compares to what can be obtained with PBS. Conclusion. It is certainly feasible to produce IMPT fields with MLC passive scattering fields. With a fully developed treatment planning system, the produced plans can be superior to traditional DS plans in terms of plan conformity and dose to organs at risk

  7. In vitro biochemical characterization of all barley endosperm starch synthases

    DEFF Research Database (Denmark)

    Cuesta-Seijo, Jose A.; Nielsen, Morten M.; Ruzanski, Christian

    2016-01-01

    Starch is the main storage polysaccharide in cereals and the major source of calories in the human diet. It is synthesized by a panel of enzymes including five classes of starch synthases (SSs). While the overall starch synthase (SS) reaction is known, the functional differences between the five SS....... Here we provide a detailed biochemical study of the activity of all five classes of SSs in barley endosperm. Each enzyme was produced recombinantly in E. coli and the properties and modes of action in vitro were studied in isolation from other SSs and other substrate modifying activities. Our results...... define the mode of action of each SS class in unprecedented detail; we analyze their substrate selection, temperature dependence and stability, substrate affinity and temporal abundance during barley development. Our results are at variance with some generally accepted ideas about starch biosynthesis...

  8. ATP Synthase, a Target for Dementia and Aging?

    Science.gov (United States)

    Larrick, James W; Larrick, Jasmine W; Mendelsohn, Andrew R

    2018-02-01

    Advancing age is the biggest risk factor for development for the major life-threatening diseases in industrialized nations accounting for >90% of deaths. Alzheimer's dementia (AD) is among the most devastating. Currently approved therapies fail to slow progression of the disease, providing only modest improvements in memory. Recently reported work describes mechanistic studies of J147, a promising therapeutic molecule previously shown to rescue the severe cognitive deficits exhibited by aged, transgenic AD mice. Apparently, J147 targets the mitochondrial alpha-F1-ATP synthase (ATP5A). Modest inhibition of the ATP synthase modulates intracellular calcium to activate AMP-activated protein kinase to inhibit mammalian target of rapamycin, a known mechanism of lifespan extension from worms to mammals.

  9. Nitric oxide synthase isoforms in spontaneous and salt hypertension

    Czech Academy of Sciences Publication Activity Database

    Hojná, Silvie; Kuneš, Jaroslav; Zicha, Josef

    2007-01-01

    Roč. 25, Suppl. 2 (2007), S 338-S 338 ISSN 0263-6352. [European Meeting on Hypertension /17./. 15.06.2007-19.06.2007, Milan] R&D Projects: GA MŠk(CZ) 1M0510 Institutional research plan: CEZ:AV0Z50110509 Keywords : nitric oxide synthase isoforms * spontaneous and salt hypertension Subject RIV: FA - Cardiovascular Diseases incl. Cardiotharic Surgery

  10. Use of linalool synthase in genetic engineering of scent production

    Science.gov (United States)

    Pichersky, Eran

    1998-01-01

    A purified S-linalool synthase polypeptide from Clarkia breweri is disclosed as is the recombinant polypeptide and nucleic acid sequences encoding the polypeptide. Also disclosed are antibodies immunoreactive with the purified peptide and with recombinant versions of the polypeptide. Methods of using the nucleic acid sequences, as well as methods of enhancing the smell and the flavor of plants expressing the nucleic acid sequences are also disclosed.

  11. Trypanosoma brucei solanesyl-diphosphate synthase localizes to the mitochondrion

    Czech Academy of Sciences Publication Activity Database

    Lai, D.-H.; Bontempi, E. J.; Lukeš, Julius

    2012-01-01

    Roč. 183, č. 2 (2012), s. 189-192 ISSN 0166-6851 R&D Projects: GA ČR(CZ) GAP305/11/2179 Institutional support: RVO:60077344 Keywords : Trypanosoma brucei * Sleeping sickness * Ubiquinone * Solanesyl-diphosphate synthase * Digitonin permeabilization * In situ tagging Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 2.734, year: 2012 http://www.sciencedirect.com/science/article/pii/S0166685112000539

  12. From bacterial to human dihydrouridine synthase: automated structure determination

    Energy Technology Data Exchange (ETDEWEB)

    Whelan, Fiona, E-mail: fiona.whelan@york.ac.uk; Jenkins, Huw T., E-mail: fiona.whelan@york.ac.uk [The University of York, Heslington, York YO10 5DD (United Kingdom); Griffiths, Samuel C. [University of Oxford, Headington, Oxford OX3 7BN (United Kingdom); Byrne, Robert T. [Ludwig-Maximilians-University Munich, Feodor-Lynen-Strasse 25, 81377 Munich (Germany); Dodson, Eleanor J.; Antson, Alfred A., E-mail: fiona.whelan@york.ac.uk [The University of York, Heslington, York YO10 5DD (United Kingdom)

    2015-06-30

    The crystal structure of a human dihydrouridine synthase, an enzyme associated with lung cancer, with 18% sequence identity to a T. maritima enzyme, has been determined at 1.9 Å resolution by molecular replacement after extensive molecular remodelling of the template. The reduction of uridine to dihydrouridine at specific positions in tRNA is catalysed by dihydrouridine synthase (Dus) enzymes. Increased expression of human dihydrouridine synthase 2 (hDus2) has been linked to pulmonary carcinogenesis, while its knockdown decreased cancer cell line viability, suggesting that it may serve as a valuable target for therapeutic intervention. Here, the X-ray crystal structure of a construct of hDus2 encompassing the catalytic and tRNA-recognition domains (residues 1–340) determined at 1.9 Å resolution is presented. It is shown that the structure can be determined automatically by phenix.mr-rosetta starting from a bacterial Dus enzyme with only 18% sequence identity and a significantly divergent structure. The overall fold of the human Dus2 is similar to that of bacterial enzymes, but has a larger recognition domain and a unique three-stranded antiparallel β-sheet insertion into the catalytic domain that packs next to the recognition domain, contributing to domain–domain interactions. The structure may inform the development of novel therapeutic approaches in the fight against lung cancer.

  13. From bacterial to human dihydrouridine synthase: automated structure determination

    International Nuclear Information System (INIS)

    Whelan, Fiona; Jenkins, Huw T.; Griffiths, Samuel C.; Byrne, Robert T.; Dodson, Eleanor J.; Antson, Alfred A.

    2015-01-01

    The crystal structure of a human dihydrouridine synthase, an enzyme associated with lung cancer, with 18% sequence identity to a T. maritima enzyme, has been determined at 1.9 Å resolution by molecular replacement after extensive molecular remodelling of the template. The reduction of uridine to dihydrouridine at specific positions in tRNA is catalysed by dihydrouridine synthase (Dus) enzymes. Increased expression of human dihydrouridine synthase 2 (hDus2) has been linked to pulmonary carcinogenesis, while its knockdown decreased cancer cell line viability, suggesting that it may serve as a valuable target for therapeutic intervention. Here, the X-ray crystal structure of a construct of hDus2 encompassing the catalytic and tRNA-recognition domains (residues 1–340) determined at 1.9 Å resolution is presented. It is shown that the structure can be determined automatically by phenix.mr-rosetta starting from a bacterial Dus enzyme with only 18% sequence identity and a significantly divergent structure. The overall fold of the human Dus2 is similar to that of bacterial enzymes, but has a larger recognition domain and a unique three-stranded antiparallel β-sheet insertion into the catalytic domain that packs next to the recognition domain, contributing to domain–domain interactions. The structure may inform the development of novel therapeutic approaches in the fight against lung cancer

  14. Dynamics of meso and thermo citrate synthases with implicit solvation

    Science.gov (United States)

    Cordeiro, J. M. M.

    The dynamics of hydration of meso and thermo citrate synthases has been investigated using the EEF1 methodology implemented with the CHARMM program. The native enzymes are composed of two identical subunits, each divided into a small and large domain. The dynamics behavior of both enzymes at 30°C and 60°C has been compared. The results of simulations show that during the hydration process, each subunit follows a different pathway of hydration, in spite of the identical sequence. The hydrated structures were compared with the crystalline structure, and the root mean square deviation (RMSD) of each residue along the trajectory was calculated. The regions with larger and smaller mobility were identified. In particular, helices belonging to the small domain are more mobile than those of the large domain. In contrast, the residues that constitute the active site show a much lower displacement compared with the crystalline structure. Hydration free energy calculations point out that Thermoplasma acidophilum citrate synthase (TCS) is more stable than chicken citrate synthase (CCS), at high temperatures. Such result has been ascribed to the higher number of superficial charges in the thermophilic homologue, which stabilizes the enzyme, while the mesophilic homologue denatures. These results are in accord with the experimental found that TCS keeps activity at temperatures farther apart from the catalysis regular temperature than the CCS.

  15. Phytochelatin synthase activity as a marker of metal pollution

    Energy Technology Data Exchange (ETDEWEB)

    Zitka, Ondrej; Krystofova, Olga; Sobrova, Pavlina [Department of Chemistry and Biochemistry, Faculty of Agronomy, Mendel University in Brno, Zemedelska 1, CZ-613 00 Brno (Czech Republic); Adam, Vojtech [Department of Chemistry and Biochemistry, Faculty of Agronomy, Mendel University in Brno, Zemedelska 1, CZ-613 00 Brno (Czech Republic); Central European Institute of Technology, Brno University of Technology, Technicka 3058/10, CZ-616 00 Brno (Czech Republic); Zehnalek, Josef; Beklova, Miroslava [Department of Chemistry and Biochemistry, Faculty of Agronomy, Mendel University in Brno, Zemedelska 1, CZ-613 00 Brno (Czech Republic); Kizek, Rene, E-mail: kizek@sci.muni.cz [Department of Chemistry and Biochemistry, Faculty of Agronomy, Mendel University in Brno, Zemedelska 1, CZ-613 00 Brno (Czech Republic); Central European Institute of Technology, Brno University of Technology, Technicka 3058/10, CZ-616 00 Brno (Czech Republic)

    2011-08-30

    Highlights: {yields} New tool for determination of phytochelatin synthase activity. {yields} The optimization of experimental condition for determination of the enzyme activity. {yields} First evaluation of K{sub m} for the enzyme. {yields} The effects of cadmium (II) not only on the activity of the enzyme but also on K{sub m}. -- Abstract: The synthesis of phytochelatins is catalyzed by {gamma}-Glu-Cys dipeptidyl transpeptidase called phytochelatin synthase (PCS). Aim of this study was to suggest a new tool for determination of phytochelatin synthase activity in the tobacco BY-2 cells treated with different concentrations of the Cd(II). After the optimization steps, an experiment on BY-2 cells exposed to different concentrations of Cd(NO{sub 3}){sub 2} for 3 days was performed. At the end of the experiment, cells were harvested and homogenized. Reduced glutathione and cadmium (II) ions were added to the cell suspension supernatant. These mixtures were incubated at 35 {sup o}C for 30 min and analysed using high performance liquid chromatography coupled with electrochemical detector (HPLC-ED). The results revealed that PCS activity rises markedly with increasing concentration of cadmium (II) ions. The lowest concentration of the toxic metal ions caused almost three fold increase in PCS activity as compared to control samples. The activity of PCS (270 fkat) in treated cells was more than seven times higher in comparison to control ones. K{sub m} for PCS was estimated as 2.3 mM.

  16. Phytochelatin synthase activity as a marker of metal pollution

    International Nuclear Information System (INIS)

    Zitka, Ondrej; Krystofova, Olga; Sobrova, Pavlina; Adam, Vojtech; Zehnalek, Josef; Beklova, Miroslava; Kizek, Rene

    2011-01-01

    Highlights: → New tool for determination of phytochelatin synthase activity. → The optimization of experimental condition for determination of the enzyme activity. → First evaluation of K m for the enzyme. → The effects of cadmium (II) not only on the activity of the enzyme but also on K m . -- Abstract: The synthesis of phytochelatins is catalyzed by γ-Glu-Cys dipeptidyl transpeptidase called phytochelatin synthase (PCS). Aim of this study was to suggest a new tool for determination of phytochelatin synthase activity in the tobacco BY-2 cells treated with different concentrations of the Cd(II). After the optimization steps, an experiment on BY-2 cells exposed to different concentrations of Cd(NO 3 ) 2 for 3 days was performed. At the end of the experiment, cells were harvested and homogenized. Reduced glutathione and cadmium (II) ions were added to the cell suspension supernatant. These mixtures were incubated at 35 o C for 30 min and analysed using high performance liquid chromatography coupled with electrochemical detector (HPLC-ED). The results revealed that PCS activity rises markedly with increasing concentration of cadmium (II) ions. The lowest concentration of the toxic metal ions caused almost three fold increase in PCS activity as compared to control samples. The activity of PCS (270 fkat) in treated cells was more than seven times higher in comparison to control ones. K m for PCS was estimated as 2.3 mM.

  17. Longevity in vivo of primary cell wall cellulose synthases.

    Science.gov (United States)

    Hill, Joseph Lee; Josephs, Cooper; Barnes, William J; Anderson, Charles T; Tien, Ming

    2018-02-01

    Our work focuses on understanding the lifetime and thus stability of the three main cellulose synthase (CESA) proteins involved in primary cell wall synthesis of Arabidopsis. It had long been thought that a major means of CESA regulation was via their rapid degradation. However, our studies here have uncovered that AtCESA proteins are not rapidly degraded. Rather, they persist for an extended time in the plant cell. Plant cellulose is synthesized by membrane-embedded cellulose synthase complexes (CSCs). The CSC is composed of cellulose synthases (CESAs), of which three distinct isozymes form the primary cell wall CSC and another set of three isozymes form the secondary cell wall CSC. We determined the stability over time of primary cell wall (PCW) CESAs in Arabidopsis thaliana seedlings, using immunoblotting after inhibiting protein synthesis with cycloheximide treatment. Our work reveals very slow turnover for the Arabidopsis PCW CESAs in vivo. Additionally, we show that the stability of all three CESAs within the PCW CSC is altered by mutations in individual CESAs, elevated temperature, and light conditions. Together, these results suggest that CESA proteins are very stable in vivo, but that their lifetimes can be modulated by intrinsic and environmental cues.

  18. The primary defect in glycogen synthase activity is not based on increased glycogen synthase kinase-3a activity in diabetic myotubes

    DEFF Research Database (Denmark)

    Gaster, Michael; Brusgaard, Klaus; Handberg, Aa.

    2004-01-01

    The mechanism responsible for the diminished activation of glycogen synthase (GS) in diabetic myotubes remains unclear, but may involve increased activity and/or expression of glycogen synthase kinase-3 (GSK-3). In myotubes established from type 2 diabetic and healthy control subjects we determined...

  19. [BIOINFORMATIC SEARCH AND PHYLOGENETIC ANALYSIS OF THE CELLULOSE SYNTHASE GENES OF FLAX (LINUM USITATISSIMUM)].

    Science.gov (United States)

    Pydiura, N A; Bayer, G Ya; Galinousky, D V; Yemets, A I; Pirko, Ya V; Podvitski, T A; Anisimova, N V; Khotyleva, L V; Kilchevsky, A V; Blume, Ya B

    2015-01-01

    A bioinformatic search of sequences encoding cellulose synthase genes in the flax genome, and their comparison to dicots orthologs was carried out. The analysis revealed 32 cellulose synthase gene candidates, 16 of which are highly likely to encode cellulose synthases, and the remaining 16--cellulose synthase-like proteins (Csl). Phylogenetic analysis of gene products of cellulose synthase genes allowed distinguishing 6 groups of cellulose synthase genes of different classes: CesA1/10, CesA3, CesA4, CesA5/6/2/9, CesA7 and CesA8. Paralogous sequences within classes CesA1/10 and CesA5/6/2/9 which are associated with the primary cell wall formation are characterized by a greater similarity within these classes than orthologous sequences. Whereas the genes controlling the biosynthesis of secondary cell wall cellulose form distinct clades: CesA4, CesA7, and CesA8. The analysis of 16 identified flax cellulose synthase gene candidates shows the presence of at least 12 different cellulose synthase gene variants in flax genome which are represented in all six clades of cellulose synthase genes. Thus, at this point genes of all ten known cellulose synthase classes are identify in flax genome, but their correct classification requires additional research.

  20. Comparison of vibrational noise, between thermal power station (T.P.S.) Jamshoro and thermal power plant (T.P.P.) Pakistan Steel Bin Qasim Karachi

    International Nuclear Information System (INIS)

    Zaheer-ud-Din Memon

    2003-01-01

    Vibrational Noise is one of the major environmental problems in Industrial Plants. The Noise study has been under taken in (Japanese Unit) Thermal Power Station (TPS) Jamshoro, which generates 250 MW since Jan: 1990 and Russian Unit Thermal Power Plant (TPP) Pak Steel Bin Qasim Karachi, generates 165 MW since 1984. The prevailing Noise has been recorded in detail; at Basement area feed pumps, Turbines, Boilers F.D.F, I.D.F and Compressor houses. Comparing these two Plants, the Noise Level found more Intensive i.e. 97- 114 dB(A) at TPP Pak Steel Bin Qasim as against 91.4 -96.3 dB(A) at TPS Jamshoro, which was even higher one in the light of ISO and other National Standards. In the light of permissible Occupational Noise exposure limits, as allowed by the ISO and other National Standards, some recommendations have been made to provide safety measures for workers against high level noise health hazards like head ache, hearing problem, irritation, accidents at work, tension, disturbance to work and so many psychological effects, along with guidelines to improve the efficiency of the plants. (author)

  1. Verification of the precision and accuracy of the TPS dosimetric calculations using the 3.0 version MIRS TECDOC-1583 of the IAEA (2008)

    International Nuclear Information System (INIS)

    Gonzalez Perez, Yelina; Rodriguez Zayas, Michael; Perez Guevara, Adrian; Sola Rodriguez, Yeline; Reyes Gonzalez, Tommy; Sanchez Zamora; Luis; Caballero, Roberto

    2009-01-01

    Radiotherapy is one of the basic therapeutic tools to treat malignant tumors. The treatment of a tumor with ionizing radiation is a continuous process with distinct stages, which is the computerized planning, being a fundamental component within this process, it is in this phase are designed and calculated the patient treatment. Systems for Radiotherapy Treatment Planning (TPS) are the tools to perform the treatment planning. The Radiotherapy Service of the Hospital Hermanos Ameijeiras acquired MIRS software version 3.0, which has potential as conventional radiation therapy planning tools and compliance with 3D, multiple imaging studies and calculation of dose according to patient data and equipment. For the complexity of these calculations and the reliability which must have the same, the software should be subjected to rigorous acceptance testing. We verify the precision and accuracy of the TPS dosimetric calculation by applying the most recent protocol of acceptance of the IAEA for these systems (2008). After implementation of the testing set is unable to verify the dose calculations are within the tolerances allowed. (Author)

  2. Eksperimentasi Model Pembelajaran Kooperatif Tipe Student Team Achievement Divisions (STAD dan Think Pair Share (TPS Terhadap Prestasi Belajar Matematika Ditinjau Dari Motivasi Berprestasi

    Directory of Open Access Journals (Sweden)

    Erny Untari

    2017-06-01

    Full Text Available The purposes of this study are to determine: (1 which one have a better mathematics achievement, students who taught by cooperative learning model STAD or TPS type. (2 which one have a better mathematics achievement, students who have high, middle, or low achievement motivation. (3 is there any interaction between learning model and achievement motivation towards mathematics achievement. This study is a quasi experimental research with 2x3 factorial design. The population of this study is all students XI Grade SMA N 1 Kwadungan in academic year 2016/2017. Sampling was done by random technique. The total of sample is 28 students, with details of 14 students for class experiment one and 14 students for class experiment two. The instruments used to collect data are test of prior knowledge in mathematics, achievement motivation questionnaire and mathematics achievement test. The trial of test instrument includes content validity, difficulty level, discrimination power, and reliability. The testing of hypothesis uses two-way analysis of variance with unequal cell. The testing of hypothesis concludes that (1 Students who taught by cooperative learning model of TPS type have better mathematics achievement than students who taught by cooperative learning model of STAD type. (2 Students who have high achievement motivation have better mathematics achievement than students who have middle and low achievement motivation, also students who have middle achievement motivation have better mathematics achievement than students who have low achievement motivation. (3 There isn’t interaction between learning model and achievement motivation towards mathematics achievement.

  3. EKSPERIMENTASI MODEL PEMBELAJARAN KOOPERATIF TIPE GROUP INVESTIGATION (GI DAN THINK PAIR AND SHARE (TPS PADA MATERI SISTEM PERSAMAAN LINEAR DITINJAU DARI SIKAP MAHASISWA TERHADAP MATEMATIKA

    Directory of Open Access Journals (Sweden)

    Swasti Maharani

    2014-02-01

    Sampel penelitian diambil dengan menggunakan teknik cluster random sampling . Data penelitian dianalisis dengan menggunakan analisis dua arah tidak seimbang varians pada tingkat signifikansi 5 %. Hasil penelitian ini menunjukkan bahwa : ( 1 model pembelajaran GI dan TPS menghasilkan prestasi belajar yang baik yang sama dalam matematika , namun kedua hasil dalam prestasi belajar yang lebih baik dalam matematika dibandingkan dengan model pembelajaran konvensional , ( 2 prestasi belajar matematika dengan sikap positif terhadap matematika adalah lebih baik dari itu dengan sikap netral dan negatif terhadap matematics , bahwa dengan sikap netral terhadap matematics adalah lebih baik dari itu dengan sikap negatif terhadap matematika , (3 pada masing-masing siswa sikap terhadap jenis matematika , GI dan model pembelajaran TPS hasil dalam pencapaian yang sama baik belajar di matematika , tapi baik mengakibatkan prestasi belajar yang lebih baik dalam matematika daripada model pembelajaran konvensional , (4 pada masing-masing model pembelajaran , prestasi belajar dengan sikap positif terhadap matematika matematika adalah lebih baik dari itu dengan netral dan sikap negatif terhadap matematics , bahwa dengan sikap netral terhadap matematics adalah lebih baik dari itu dengan sikap negatif terhadap matematika.

  4. Structure of the human beta-ketoacyl [ACP] synthase from the mitochondrial type II fatty acid synthase

    DEFF Research Database (Denmark)

    Christensen, Caspar Elo; Kragelund, Birthe B; von Wettstein-Knowles, Penny

    2007-01-01

    Two distinct ways of organizing fatty acid biosynthesis exist: the multifunctional type I fatty acid synthase (FAS) of mammals, fungi, and lower eukaryotes with activities residing on one or two polypeptides; and the dissociated type II FAS of prokaryotes, plastids, and mitochondria with individual...... activities encoded by discrete genes. The beta-ketoacyl [ACP] synthase (KAS) moiety of the mitochondrial FAS (mtKAS) is targeted by the antibiotic cerulenin and possibly by the other antibiotics inhibiting prokaryotic KASes: thiolactomycin, platensimycin, and the alpha-methylene butyrolactone, C75. The high...... degree of structural similarity between mitochondrial and prokaryotic KASes complicates development of novel antibiotics targeting prokaryotic KAS without affecting KAS domains of cytoplasmic FAS. KASes catalyze the C(2) fatty acid elongation reaction using either a Cys-His-His or Cys-His-Asn catalytic...

  5. Cloning and characterization of indole synthase (INS) and a putative tryptophan synthase α-subunit (TSA) genes from Polygonum tinctorium.

    Science.gov (United States)

    Jin, Zhehao; Kim, Jin-Hee; Park, Sang Un; Kim, Soo-Un

    2016-12-01

    Two cDNAs for indole-3-glycerol phosphate lyase homolog were cloned from Polygonum tinctorium. One encoded cytosolic indole synthase possibly in indigoid synthesis, whereas the other encoded a putative tryptophan synthase α-subunit. Indigo is an old natural blue dye produced by plants such as Polygonum tinctorium. Key step in plant indigoid biosynthesis is production of indole by indole-3-glycerol phosphate lyase (IGL). Two tryptophan synthase α-subunit (TSA) homologs, PtIGL-short and -long, were isolated by RACE PCR from P. tinctorium. The genome of the plant contained two genes coding for IGL. The short and the long forms, respectively, encoded 273 and 316 amino acid residue-long proteins. The short form complemented E. coli ΔtnaA ΔtrpA mutant on tryptophan-depleted agar plate signifying production of free indole, and thus was named indole synthase gene (PtINS). The long form, either intact or without the transit peptide sequence, did not complement the mutant and was tentatively named PtTSA. PtTSA was delivered into chloroplast as predicted by 42-residue-long targeting sequence, whereas PtINS was localized in cytosol. Genomic structure analysis suggested that a TSA duplicate acquired splicing sites during the course of evolution toward PtINS so that the targeting sequence-containing pre-mRNA segment was deleted as an intron. PtINS had about two to fivefolds higher transcript level than that of PtTSA, and treatment of 2,1,3-benzothiadiazole caused the relative transcript level of PtINS over PtTSA was significantly enhanced in the plant. The results indicate participation of PtINS in indigoid production.

  6. Characterization of Space Shuttle Thermal Protection System (TPS) Materials for Return-to-Flight following the Shuttle Columbia Accident Investigation

    Science.gov (United States)

    Wingard, Doug

    2006-01-01

    During the Space Shuttle Columbia Accident Investigation, it was determined that a large chunk of polyurethane insulating foam (= 1.67 lbs) on the External Tank (ET) came loose during Columbia's ascent on 2-1-03. The foam piece struck some of the protective Reinforced Carbon-Carbon (RCC) panels on the leading edge of Columbia's left wing in the mid-wing area. This impact damaged Columbia to the extent that upon re-entry to Earth, superheGed air approaching 3,000 F caused the vehicle to break up, killing all seven astronauts on board. A paper after the Columbia Accident Investigation highlighted thermal analysis testing performed on External Tank TPS materials (1). These materials included BX-250 (now BX-265) rigid polyurethane foam and SLA-561 Super Lightweight Ablator (highly-filled silicone rubber). The large chunk of foam from Columbia originated fiom the left bipod ramp of the ET. The foam in this ramp area was hand-sprayed over the SLA material and various fittings, allowed to dry, and manually shaved into a ramp shape. In Return-to-Flight (RTF) efforts following Columbia, the decision was made to remove the foam in the bipod ramp areas. During RTF efforts, further thermal analysis testing was performed on BX-265 foam by DSC and DMA. Flat panels of foam about 2-in. thick were sprayed on ET tank material (aluminum alloys). The DSC testing showed that foam material very close to the metal substrate cured more slowly than bulk foam material. All of the foam used on the ET is considered fully cured about 21 days after it is sprayed. The RTF culminated in the successful launch of Space Shuttle Discovery on 7-26-05. Although the flight was a success, there was another serious incident of foam loss fiom the ET during Shuttle ascent. This time, a rather large chunk of BX-265 foam (= 0.9 lbs) came loose from the liquid hydrogen (LH2) PAL ramp, although the foam did not strike the Shuttle Orbiter containing the crew. DMA testing was performed on foam samples taken fiom

  7. Long-term stability in biomass and production of terpene indole alkaloids by hairy root culture of Rauvolfia serpentina and cost approximation to endorse commercial realism.

    Science.gov (United States)

    Pandey, Pallavi; Kaur, Ranjeet; Singh, Sailendra; Chattopadhyay, Sunil Kumar; Srivastava, Santosh Kumar; Banerjee, Suchitra

    2014-07-01

    The effect of 6 years of cultivation and use of table-sugar (TS) on the biomass/terpene alkaloid productivities and rol gene expression were studied in a hairy root (HR) clone of Rauvolfia serpentina. The media cost could be reduced >94 % by replacing sucrose (SUC) with TS—an unexplored avenue for HR cultivation. The overall productivities increased over long-term cultivation with sugar proving superior to SUC for biomass (24.4 ± 2.11 g/l DW after 40 days to 17.31 % higher) and reserpine (0.094 ± 0.008 % DW after 60 days to 193.8 % more) production. The latter however revealed comparatively better yields concerning ajmaline (0.507 ± 0.048 % DW after 60 days to 61.98 % higher) and yohimbine (0.628 ± 0.062 % DW after 60 days to 38.32 % higher), respectively. PCR amplification of rol genes confirmed long-term expression stability.

  8. Enhancement of vascular targeting by inhibitors of nitric oxide synthase

    International Nuclear Information System (INIS)

    Davis, Peter D.; Tozer, Gillian M.; Naylor, Matthew A.; Thomson, Peter; Lewis, Gemma; Hill, Sally A.

    2002-01-01

    Purpose: This study investigates the enhancement of the vascular targeting activity of the tubulin-binding agent combretastatin A4 phosphate (CA4P) by various inhibitors of nitric oxide synthases. Methods and Materials: The syngeneic tumors CaNT and SaS growing in CBA mice were used for this study. Reduction in perfused vascular volume was measured by injection of Hoechst 33342 24 h after drug administration. Necrosis (hematoxylin and eosin stain) was assessed also at 24 h after treatment. Combretastatin A4 phosphate was synthesized by a modification of the published procedure and the nitric oxide synthase inhibitors L-NNA, L-NMMA, L-NIO, L-NIL, S-MTC, S-EIT, AMP, AMT, and L-TC, obtained from commercial sources. Results: A statistically significant augmentation of the reduction in perfused vascular volume by CA4P in the CaNT tumor was observed with L-NNA, AMP, and AMT. An increase in CA4P-induced necrosis in the same tumor achieved significance with L-NNA, L-NMMA, L-NIL, and AMT. CA4P induced little necrosis in the SaS tumor, but combination with the inhibitors L-NNA, L-NMMA, L-NIO, S-EIT, and L-TC was effective. Conclusions: Augmentation of CA4P activity by nitric oxide synthase inhibitors of different structural classes supports a nitric oxide-related mechanism for this effect. L-NNA was the most effective inhibitor studied

  9. Incorporation of phosphate into glycogen by glycogen synthase.

    Science.gov (United States)

    Contreras, Christopher J; Segvich, Dyann M; Mahalingan, Krishna; Chikwana, Vimbai M; Kirley, Terence L; Hurley, Thomas D; DePaoli-Roach, Anna A; Roach, Peter J

    2016-05-01

    The storage polymer glycogen normally contains small amounts of covalently attached phosphate as phosphomonoesters at C2, C3 and C6 atoms of glucose residues. In the absence of the laforin phosphatase, as in the rare childhood epilepsy Lafora disease, the phosphorylation level is elevated and is associated with abnormal glycogen structure that contributes to the pathology. Laforin therefore likely functions in vivo as a glycogen phosphatase. The mechanism of glycogen phosphorylation is less well-understood. We have reported that glycogen synthase incorporates phosphate into glycogen via a rare side reaction in which glucose-phosphate rather than glucose is transferred to a growing polyglucose chain (Tagliabracci et al. (2011) Cell Metab13, 274-282). We proposed a mechanism to account for phosphorylation at C2 and possibly at C3. Our results have since been challenged (Nitschke et al. (2013) Cell Metab17, 756-767). Here we extend the evidence supporting our conclusion, validating the assay used for the detection of glycogen phosphorylation, measurement of the transfer of (32)P from [β-(32)P]UDP-glucose to glycogen by glycogen synthase. The (32)P associated with the glycogen fraction was stable to ethanol precipitation, SDS-PAGE and gel filtration on Sephadex G50. The (32)P-signal was not affected by inclusion of excess unlabeled UDP before analysis or by treatment with a UDPase, arguing against the signal being due to contaminating [β-(32)P]UDP generated in the reaction. Furthermore, [(32)P]UDP did not bind non-covalently to glycogen. The (32)P associated with glycogen was released by laforin treatment, suggesting that it was present as a phosphomonoester. The conclusion is that glycogen synthase can mediate the introduction of phosphate into glycogen, thereby providing a possible mechanism for C2, and perhaps C3, phosphorylation. Copyright © 2016 Elsevier Inc. All rights reserved.

  10. Microsomal prostaglandin E synthase-1 in rheumatic diseases

    Directory of Open Access Journals (Sweden)

    Marina eKorotkova

    2011-01-01

    Full Text Available Microsomal prostaglandin E synthase-1 (mPGES-1 is a well recognized target for the development of novel anti-inflammatory drugs that can reduce symptoms of inflammation in rheumatic diseases and other inflammatory conditions. In this review, we focus on mPGES-1 in rheumatic diseases with the aim to cover the most recent advances in the understanding of mPGES-1 in rheumatoid arthritis, osteoarthritis and inflammatory myopathies. Novel findings regarding regulation of mPGES1 cell expression as well as enzyme inhibitors are also summarized.

  11. Nitric oxide synthase expression and enzymatic activity in multiple sclerosis

    DEFF Research Database (Denmark)

    Broholm, H; Andersen, B; Wanscher, B

    2004-01-01

    We used post-mortem magnetic resonance imaging (MRI) guidance to obtain paired biopsies from the brains of four patients with clinical definite multiple sclerosis (MS). Samples were analyzed for the immunoreactivity (IR) of the three nitric oxide (NO) synthase isoforms [inducible, neuronal......NOS expressing cells in active lesions. NOS IR expressing cells were widely distributed in plaques, in white and gray matter that appeared normal macroscopically, and on MR. Endothelial NOS (eNOS) was highly expressed in intraparenchymal vascular endothelial cells of MS patients. A control group matched for age...

  12. Fatty acid synthase inhibitors isolated from Punica granatum L

    Energy Technology Data Exchange (ETDEWEB)

    Jiang, He-Zhong [School of Life Science and Engineering, Southwest Jiaotong University, Chengdu, (China); Ma, Qing-Yun; Liang, Wen-Juan; Huang, Sheng-Zhuo; Dai, Hao-Fu; Wang, Peng-Cheng; Zhao, You-Xing, E-mail: zhaoyx1011@163.com [Institute of Tropical Bioscience and Biotechnology, Chinese Academy of Tropical Agricultural Sciences, Haikou (China); Fan, Hui-Jin; Ma, Xiao-Feng, E-mail: maxiaofeng@gucas.ac.cn [College of Life Sciences, Graduate University of Chinese Academy of Sciences, Beijing (China)

    2012-05-15

    The aim of this work is the isolation of fatty acid synthase (FAS) inhibitors from the ethyl acetate extracts of fruit peels of Punica granatum L. Bioassay-guided chemical investigation of the fruit peels resulted in the isolation of seventeen compounds mainly including triterpenoids and phenolic compounds, from which one new oleanane-type triterpene (punicaone) along with fourteen known compounds were isolated for the first time from this plant. Seven isolates were evaluated for inhibitory activities of FAS and two compounds showed to be active. Particularly, flavogallonic acid exhibited strong FAS inhibitory activity with IC{sub 50} value of 10.3 {mu}mol L{sup -1}. (author)

  13. Fatty acid synthase inhibitors isolated from Punica granatum L

    International Nuclear Information System (INIS)

    Jiang, He-Zhong; Ma, Qing-Yun; Liang, Wen-Juan; Huang, Sheng-Zhuo; Dai, Hao-Fu; Wang, Peng-Cheng; Zhao, You-Xing; Fan, Hui-Jin; Ma, Xiao-Feng

    2012-01-01

    The aim of this work is the isolation of fatty acid synthase (FAS) inhibitors from the ethyl acetate extracts of fruit peels of Punica granatum L. Bioassay-guided chemical investigation of the fruit peels resulted in the isolation of seventeen compounds mainly including triterpenoids and phenolic compounds, from which one new oleanane-type triterpene (punicaone) along with fourteen known compounds were isolated for the first time from this plant. Seven isolates were evaluated for inhibitory activities of FAS and two compounds showed to be active. Particularly, flavogallonic acid exhibited strong FAS inhibitory activity with IC 50 value of 10.3 μmol L -1 . (author)

  14. Perbedaan Keterampilan Metakognitif dan Motivasi Siswa Putra dan Putri Kelas X SMAN Di Kota Malang Melalui Strategi Pembelajaran Reading Questioning and Answering (RQA Dipadu Think Pair Share (TPS

    Directory of Open Access Journals (Sweden)

    Hindun Syarifah

    2016-03-01

    Full Text Available Student diversity issue in school is one of the subjects in educational research. The quite apparent problem of student diversity in the school is gender differences. Male and female are different in some ways. Several studies have revealed the influence of gender differences on a wide range of students' abilities. Mahanal (2011 revealed that there was an effect of gender differences on metacognition skills and critical thinking abilities of high school students in Malang city. This research is aimed to determine the metacognition skills and motivation differences of male and female student on the biology subject through the implementation of Reading Questioning and Answering (RQA combined with Think Pair Share (TPS learning strategies. This research used quasy-experiment of pre test- post test nonequivalent group design. The research populations were all of

  15. Molecular cloning and functional expression of geranylgeranyl pyrophosphate synthase from Coleus forskohlii Briq

    Directory of Open Access Journals (Sweden)

    Kawamukai Makoto

    2004-11-01

    Full Text Available Abstract Background Isopentenyl diphosphate (IPP, a common biosynthetic precursor to the labdane diterpene forskolin, has been biosynthesised via a non-mevalonate pathway. Geranylgeranyl diphosphate (GGPP synthase is an important branch point enzyme in terpenoid biosynthesis. Therefore, GGPP synthase is thought to be a key enzyme in biosynthesis of forskolin. Herein we report the first confirmation of the GGPP synthase gene in Coleus forskohlii Briq. Results The open reading frame for full-length GGPP synthase encodes a protein of 359 amino acids, in which 1,077 nucleotides long with calculated molecular mass of 39.3 kDa. Alignments of C. forskohlii GGPP synthase amino acid sequences revealed high homologies with other plant GGPP synthases. Several highly conserved regions, including two aspartate-rich motifs were identified. Transient expression of the N-terminal region of C. forskohlii GGPP synthase-GFP fusion protein in tobacco cells demonstrated subcellular localization in the chloroplast. Carotenoid production was observed in Escherichia coli harboring pACCAR25ΔcrtE from Erwinia uredovora and plasmid carrying C. forskohlii GGPP synthase. These results suggested that cDNA encoded functional GGPP synthase. Furthermore, C. forskohlii GGPP synthase expression was strong in leaves, decreased in stems and very little expression was observed in roots. Conclusion This investigation proposed that forskolin was synthesised via a non-mevalonate pathway. GGPP synthase is thought to be involved in the biosynthesis of forskolin, which is primarily synthesised in the leaves and subsequently accumulates in the stems and roots.

  16. Glycogen synthase from the parabasalian parasite Trichomonas vaginalis: An unusual member of the starch/glycogen synthase family.

    Science.gov (United States)

    Wilson, Wayne A; Pradhan, Prajakta; Madhan, Nayasha; Gist, Galen C; Brittingham, Andrew

    2017-07-01

    Trichomonas vaginalis, a parasitic protist, is the causative agent of the common sexually-transmitted infection trichomoniasis. The organism has long been known to synthesize substantial glycogen as a storage polysaccharide, presumably mobilizing this compound during periods of carbohydrate limitation, such as might be encountered during transmission between hosts. However, little is known regarding the enzymes of glycogen metabolism in T. vaginalis. We had previously described the identification and characterization of two forms of glycogen phosphorylase in the organism. Here, we measure UDP-glucose-dependent glycogen synthase activity in cell-free extracts of T. vaginalis. We then demonstrate that the TVAG_258220 open reading frame encodes a glycosyltransferase that is presumably responsible for this synthetic activity. We show that expression of TVAG_258220 in a yeast strain lacking endogenous glycogen synthase activity is sufficient to restore glycogen accumulation. Furthermore, when TVAG_258220 is expressed in bacteria, the resulting recombinant protein has glycogen synthase activity in vitro, transferring glucose from either UDP-glucose or ADP-glucose to glycogen and using both substrates with similar affinity. This protein is also able to transfer glucose from UDP-glucose or ADP-glucose to maltose and longer oligomers of glucose but not to glucose itself. However, with these substrates, there is no evidence of processivity and sugar transfer is limited to between one and three glucose residues. Taken together with our earlier work on glycogen phosphorylase, we are now well positioned to define both how T. vaginalis synthesizes and utilizes glycogen, and how these processes are regulated. Copyright © 2017 Elsevier B.V. and Société Française de Biochimie et Biologie Moléculaire (SFBBM). All rights reserved.

  17. PENERAPAN MODEL PEMBELAJARAN KOOPERATIF TIPE THINK PAIR SHARE (TPS PADA KOMPETENSI DASAR SURAT MENYURAT UNTUK MENINGKATKAN AKTIVITAS BELAJAR SISWA KELAS XI DI SMK PGRI 2 SALATIGA

    Directory of Open Access Journals (Sweden)

    Fatmawati Nur Fadhilah

    2013-11-01

    Full Text Available Tujuan penelitian ini adalah untuk meningkatkan aktivitas belajar siswa kelas XI di SMK PGRI 2 Salatiga. Subjek dalam penelitian ini adalah kelas XI AP-3 yang berjumlah 31 siswa. Penelitian ini dilaksanakan dalam 2 (dua siklus. Kegiatan dalam setiap siklus meliputi kegiatan perencanaan, pelaksanaan, observasi, dan refleksi. Hasil penelitian menunjukkan bahwa aktivitas belajar siswa pada siklus I diperoleh total skor 36 dari 10 aspek yang diamati, dan hasil perhitungan aktivitas belajar siswa diperoleh persentase sebesar 72%, nilai tersebut dalam kategori tinggi. Pada penilaian aktivitas belajar siswa siklus II meningkat dengan total skor 42 dari 10 aspek yang diamati, dan hasil perhitungan aktivitas belajar siswa diperoleh persentase sebesar 84%, nilai tersebut dalam kategori sangat tinggi. Berdasarkan hasil penelitian tersebut, dapat disimpulkan bahwa ada peningkatan aktivitas siswa kelas XI Administrasi Perkantoran 3 pada Kompetensi Dasar Surat Menyurat melalui model pembelajaran kooperatif tipe Think Pair Share (TPS. The purpose of this research is to improve students' learning activities in class XI SMK PGRI 2 Salatiga. Subjects in this study were class XI AP-3 is 31 students. This study was conducted in two (2 cycles. Activities in each cycle includes planning, implementation, observation, and reflection. The results showed that students' learning activities in the first cycle obtained a total score of 36 out of 10 aspects were observed, and the calculation results obtained by the percentage of student learning activity by 72%, the value is in the high category. In the assessment of student learning activities second cycle increased by a total score of 42 out of 10 aspects are observed, and the calculation results obtained by the percentage of students' learning activities by 84%, the value is in the very high category. Based on these results, we can conclude that there is an increase in the activity of the students of class XI

  18. Stereochemical course of enzyme-catalyzed aminopropyl transfer: spermidine synthase

    International Nuclear Information System (INIS)

    Kullberg, D.W.; Orr, G.R.; Coward, J.K.

    1986-01-01

    The R and S enantionmers of S-adenosyl-3-[ 2 H]3-(methylthio)-1-propylamine (decarboxylated S-adenosylmethionine), previously synthesized in this laboratory, were incubated with [1,4- 2 H 4 ]-putrescine in the presence of spermidine synthase from E. coli. The resulting chiral [ 2 H 5 ]spermidines were isolated and converted to their N 1 ,N 7 -dibocspermidine-N 4 -(1S,4R)-camphanamides. The derivatives were analyzed by 500 MHz 1 H-NMR and the configuration of the chiral center assigned by correlation with the spectra of synthetic chiral [ 2 H 3 ]dibocspermidine camphanamide standards. The enzyme-catalyzed aminopropyl transfer was shown to occur with net retention of configuration, indicative of a double-displacement mechanism. This result concurs with that of a previous steady-state kinetics study of spermidine synthase isolated from E. coli, but contradicts the single-displacement mechanism suggested by a stereochemical analysis of chiral spermidines biosynthesized in E. coli treated with chirally deuterated methionines. It also indicates that this aminopropyltransferase is mechanistically distinct from the methyltransferases, which have been shown to act via a single-displacement mechanism (net inversion at -CH 3 ) in all cases studied to date

  19. In Vitro Biochemical Characterization of All Barley Endosperm Starch Synthases

    Directory of Open Access Journals (Sweden)

    Jose Antonio Cuesta-Seijo

    2016-01-01

    Full Text Available Starch is the main storage polysaccharide in cereals and the major source of calories in the human diet. It is synthesized by a panel of enzymes including five classes of starch synthases (SSs. While the overall starch synthase (SS reaction is known, the functional differences between the five SS classes are poorly understood. Much of our knowledge comes from analyzing mutant plants with altered SS activities, but the resulting data are often difficult to interpret as a result of pleitropic effects, competition between enzymes, overlaps in enzyme activity and disruption of multi-enzyme complexes. Here we provide a detailed biochemical study of the activity of all five classes of SSs in barley endosperm. Each enzyme was produced recombinantly in E. coli and the properties and modes of action in vitro were studied in isolation from other SSs and other substrate modifying activities. Our results define the mode of action of each SS class in unprecedented detail; we analyze their substrate selection, temperature dependence and stability, substrate affinity and temporal abundance during barley development. Our results are at variance with some generally accepted ideas about starch biosynthesis and might lead to the reinterpretation of results obtained in planta. In particular, they indicate that granule bound SS is capable of processive action even in the absence of a starch matrix, that SSI has no elongation limit, and that SSIV, believed to be critical for the initiation of starch granules, has maltoligosaccharides and not polysaccharides as its preferred substrates.

  20. Modulation of hyaluronan synthase activity in cellular membrane fractions.

    Science.gov (United States)

    Vigetti, Davide; Genasetti, Anna; Karousou, Evgenia; Viola, Manuela; Clerici, Moira; Bartolini, Barbara; Moretto, Paola; De Luca, Giancarlo; Hascall, Vincent C; Passi, Alberto

    2009-10-30

    Hyaluronan (HA), the only non-sulfated glycosaminoglycan, is involved in morphogenesis, wound healing, inflammation, angiogenesis, and cancer. In mammals, HA is synthesized by three homologous HA synthases, HAS1, HAS2, and HAS3, that polymerize the HA chain using UDP-glucuronic acid and UDP-N-acetylglucosamine as precursors. Since the amount of HA is critical in several pathophysiological conditions, we developed a non-radioactive assay for measuring the activity of HA synthases (HASs) in eukaryotic cells and addressed the question of HAS activity during intracellular protein trafficking. We prepared three cellular fractions: plasma membrane, cytosol (containing membrane proteins mainly from the endoplasmic reticulum and Golgi), and nuclei. After incubation with UDP-sugar precursors, newly synthesized HA was quantified by polyacrylamide gel electrophoresis of fluorophore-labeled saccharides and high performance liquid chromatography. This new method measured HAS activity not only in the plasma membrane fraction but also in the cytosolic membranes. This new technique was used to evaluate the effects of 4-methylumbeliferone, phorbol 12-myristate 13-acetate, interleukin 1beta, platelet-derived growth factor BB, and tunicamycin on HAS activities. We found that HAS activity can be modulated by post-translational modification, such as phosphorylation and N-glycosylation. Interestingly, we detected a significant increase in HAS activity in the cytosolic membrane fraction after tunicamycin treatment. Since this compound is known to induce HA cable structures, this result links HAS activity alteration with the capability of the cell to promote HA cable formation.

  1. Glycogen synthase kinase 3: more than a namesake.

    Science.gov (United States)

    Rayasam, Geetha Vani; Tulasi, Vamshi Krishna; Sodhi, Reena; Davis, Joseph Alex; Ray, Abhijit

    2009-03-01

    Glycogen synthase kinase 3 (GSK3), a constitutively acting multi-functional serine threonine kinase is involved in diverse physiological pathways ranging from metabolism, cell cycle, gene expression, development and oncogenesis to neuroprotection. These diverse multiple functions attributed to GSK3 can be explained by variety of substrates like glycogen synthase, tau protein and beta catenin that are phosphorylated leading to their inactivation. GSK3 has been implicated in various diseases such as diabetes, inflammation, cancer, Alzheimer's and bipolar disorder. GSK3 negatively regulates insulin-mediated glycogen synthesis and glucose homeostasis, and increased expression and activity of GSK3 has been reported in type II diabetics and obese animal models. Consequently, inhibitors of GSK3 have been demonstrated to have anti-diabetic effects in vitro and in animal models. However, inhibition of GSK3 poses a challenge as achieving selectivity of an over achieving kinase involved in various pathways with multiple substrates may lead to side effects and toxicity. The primary concern is developing inhibitors of GSK3 that are anti-diabetic but do not lead to up-regulation of oncogenes. The focus of this review is the recent advances and the challenges surrounding GSK3 as an anti-diabetic therapeutic target.

  2. Polyketide synthases from poison hemlock (Conium maculatum L.).

    Science.gov (United States)

    Hotti, Hannu; Seppänen-Laakso, Tuulikki; Arvas, Mikko; Teeri, Teemu H; Rischer, Heiko

    2015-11-01

    Coniine is a toxic alkaloid, the biosynthesis of which is not well understood. A possible route, supported by evidence from labelling experiments, involves a polyketide formed by the condensation of one acetyl-CoA and three malonyl-CoAs catalysed by a polyketide synthase (PKS). We isolated PKS genes or their fragments from poison hemlock (Conium maculatum L.) by using random amplification of cDNA ends (RACE) and transcriptome analysis, and characterized three full-length enzymes by feeding different starter-CoAs in vitro. On the basis of our in vitro experiments, two of the three characterized PKS genes in poison hemlock encode chalcone synthases (CPKS1 and CPKS2), and one encodes a novel type of PKS (CPKS5). We show that CPKS5 kinetically favours butyryl-CoA as a starter-CoA in vitro. Our results suggest that CPKS5 is responsible for the initiation of coniine biosynthesis by catalysing the synthesis of the carbon backbone from one butyryl-CoA and two malonyl-CoAs. © 2015 FEBS.

  3. Amaryllidaceae Alkaloids as Potential Glycogen Synthase Kinase-3β Inhibitors

    Directory of Open Access Journals (Sweden)

    Daniela Hulcová

    2018-03-01

    Full Text Available Glycogen synthase kinase-3β (GSK-3β is a multifunctional serine/threonine protein kinase that was originally identified as an enzyme involved in the control of glycogen metabolism. It plays a key role in diverse physiological processes including metabolism, the cell cycle, and gene expression by regulating a wide variety of well-known substances like glycogen synthase, tau-protein, and β-catenin. Recent studies have identified GSK-3β as a potential therapeutic target in Alzheimer´s disease, bipolar disorder, stroke, more than 15 types of cancer, and diabetes. GSK-3β is one of the most attractive targets for medicinal chemists in the discovery, design, and synthesis of new selective potent inhibitors. In the current study, twenty-eight Amaryllidaceae alkaloids of various structural types were studied for their potency to inhibit GSK-3β. Promising results have been demonstrated by alkaloids of the homolycorine-{9-O-demethylhomolycorine (IC50 = 30.00 ± 0.71 µM, masonine (IC50 = 27.81 ± 0.01 μM}, and lycorine-types {caranine (IC50 = 30.75 ± 0.04 μM}.

  4. The crystal structure of human GDP-L-fucose synthase.

    Science.gov (United States)

    Zhou, Huan; Sun, Lihua; Li, Jian; Xu, Chunyan; Yu, Feng; Liu, Yahui; Ji, Chaoneng; He, Jianhua

    2013-09-01

    Human GDP-l-fucose synthase, also known as FX protein, synthesizes GDP-l-fucose from its substrate GDP-4-keto-6-deoxy-d-mannose. The reaction involves epimerization at both C-3 and C-5 followed by an NADPH-dependent reduction of the carbonyl at C-4. In this paper, the first crystal structure of human FX protein was determined at 2.37 Å resolution. The asymmetric unit of the crystal structure contains four molecules which form two homodimers. Each molecule consists of two domains, a Rossmann-fold NADPH-binding motif and a carboxyl terminal domain. Compared with the Escherichia coli GDP-l-fucose synthase, the overall structures of these two enzymes have four major differences. There are four loops in the structure of human FX protein corresponding to two α-helices and two β-sheets in that of the E. coli enzyme. Besides, there are seven different amino acid residues binding with NAPDH comparing human FX protein with that from E. coli. The structure of human FX reveals the key catalytic residues and could be useful for the design of drugs for the treatment of inflammation, auto-immune diseases, and possibly certain types of cancer.

  5. Prostaglandin E(2) synthase inhibition as a therapeutic target.

    Science.gov (United States)

    Iyer, Jitesh P; Srivastava, Punit K; Dev, Rishabh; Dastidar, Sunanda G; Ray, Abhijit

    2009-07-01

    Most NSAIDs function by inhibiting biosynthesis of PGE(2) by inhibition of COX-1 and/or COX-2. Since COX-1 has a protective function in the gastro-intestinal tract (GIT), non-selective inhibition of both cycloxy genases leads to moderate to severe gastro-intestinal intolerance. Attempts to identify selective inhibitors of COX-2, led to the identification of celecoxib and rofecoxib. However, long-term use of these drugs has serious adverse effects of sudden myocardial infarction and thrombosis. Drug-mediated imbalance in the levels of prostaglandin I(2) (PGI(2)) and thromboxane A(2) (TXA(2)) with a bias towards TXA(2) may be the primary reason for these events. This resulted in the drugs being withdrawn from the market, leaving a need for an effective and safe anti-inflammatory drug. Recently, the focus of research has shifted to enzymes downstream of COX in the prosta glandin biosynthetic pathway such as prostaglandin E(2) synthases. Microsomal prostaglandin E(2) synthase-1 (mPGES-1) specifically isomerizes PGH(2) to PGE(2), under inflammatory conditions. In this review, we examine the biology of mPGES-1 and its role in disease. Progress in designing molecules that can selectively inhibit mPGES-1 is reviewed. mPGES-1 has the potential to be a target for anti-inflammatory therapy, devoid of adverse GIT and cardiac effects and warrants further investigation.

  6. Chromosomal localization of the human and mouse hyaluronan synthase genes

    Energy Technology Data Exchange (ETDEWEB)

    Spicer, A.P.; McDonald, J.A. [Mayo Clinic Scottsdale, AZ (United States); Seldin, M.F. [Univ. of California Davis, CA (United States)] [and others

    1997-05-01

    We have recently identified a new vertebrate gene family encoding putative hyaluronan (HA) synthases. Three highly conserved related genes have been identified, designated HAS1, HAS2, and HAS3 in humans and Has1, Has2, and Has3 in the mouse. All three genes encode predicted plasma membrane proteins with multiple transmembrane domains and approximately 25% amino acid sequence identity to the Streptococcus pyogenes HA synthase, HasA. Furthermore, expression of any one HAS gene in transfected mammalian cells leads to high levels of HA biosynthesis. We now report the chromosomal localization of the three HAS genes in human and in mouse. The genes localized to three different positions within both the human and the mouse genomes. HAS1 was localized to the human chromosome 19q13.3-q13.4 boundary and Has1 to mouse Chr 17. HAS2 was localized to human chromosome 8q24.12 and Has2 to mouse Chr 15. HAS3 was localized to human chromosome 16q22.1 and Has3 to mouse Chr 8. The map position for HAS1 reinforces the recently reported relationship between a small region of human chromosome 19q and proximal mouse chromosome 17. HAS2 mapped outside the predicted critical region delineated for the Langer-Giedion syndrome and can thus be excluded as a candidate gene for this genetic syndrome. 33 refs., 2 figs.

  7. Selectivity of the surface binding site (SBS) on barley starch synthase I

    DEFF Research Database (Denmark)

    Wilkens, Casper; Cuesta-Seijo, Jose A.; Palcic, Monica

    2014-01-01

    Starch synthase I (SSI) from various sources has been shown to preferentially elongate branch chains of degree of polymerisation (DP) from 6–7 to produce chains of DP 8–12. In the recently determined crystal structure of barley starch synthase I (HvSSI) a so-called surface binding site (SBS) was ...

  8. Inhibition of the ATP Synthase Eliminates the Intrinsic Resistance of Staphylococcus aureus towards Polymyxins

    DEFF Research Database (Denmark)

    Vestergaard, Martin; Nøhr-Meldgaard, Katrine; Bojer, Martin Saxtorph

    2017-01-01

    , linezolid, daptomycin, and oxacillin were unchanged. ATP synthase activity is known to be inhibited by oligomycin A, and the presence of this compound increased polymyxin B-mediated killing of S. aureus Our results demonstrate that the ATP synthase contributes to intrinsic resistance of S. aureus towards...

  9. Creation of a high-amylose durum wheat through mutagenesis of starch synthase II (SSIIa)

    Science.gov (United States)

    In cereal seeds mutations in one or more starch synthases lead to decreased amylopectin and increased amylose content. Here, the impact of starch synthase IIa (SSIIa or SGP-1) mutations upon durum starch was investigated. A screen of durum accessions identified two lines lacking SGP-A1, the A geno...

  10. Cytidine triphosphate synthase activity and mRNA expression in normal human blood cells

    NARCIS (Netherlands)

    Verschuur, A. C.; van Gennip, A. H.; Muller, E. J.; Voûte, P. A.; Vreken, P.; van Kuilenburg, A. B.

    1999-01-01

    Cytidine triphosphate (CTP) synthase is one of the key enzymes in pyrimidine nucleotide anabolic pathways. The activity of this enzyme is elevated in various malignancies including acute lymphocytic leukemia (ALL). In this study we investigated the activity of CTP synthase in various human blood

  11. Cloning and sequence analysis of putative type II fatty acid synthase ...

    Indian Academy of Sciences (India)

    Prakash

    Cloning and sequence analysis of putative type II fatty acid synthase genes from Arachis hypogaea L. ... acyl carrier protein (ACP), malonyl-CoA:ACP transacylase, β-ketoacyl-ACP .... Helix II plays a dominant role in the interaction ... main distinguishing features of plant ACPs in plastids and ..... synthase component; J. Biol.

  12. Reduced methylation of the thromboxane synthase gene is correlated with its increased vascular expression in preeclampsia.

    Science.gov (United States)

    Mousa, Ahmad A; Strauss, Jerome F; Walsh, Scott W

    2012-06-01

    Preeclampsia is characterized by increased thromboxane and decreased prostacyclin levels, which predate symptoms, and can explain some of the clinical manifestations of preeclampsia, including hypertension and thrombosis. In this study, we examined DNA methylation of the promoter region of the thromboxane synthase gene (TBXAS1) and the expression of thromboxane synthase in systemic blood vessels of normal pregnant and preeclamptic women. Thromboxane synthase is responsible for the synthesis of thromboxane A(2), a potent vasoconstrictor and activator of platelets. We also examined the effect of experimentally induced DNA hypomethylation on the expression of thromboxane synthase in a neutrophil-like cell line (HL-60 cells) and in cultured vascular smooth muscle and endothelial cells. We found that DNA methylation of the TBXAS1 promoter was decreased and thromboxane synthase expression was increased in omental arteries of preeclamptic women as compared with normal pregnant women. Increased thromboxane synthase expression was observed in vascular smooth muscles cells, endothelial cells, and infiltrating neutrophils. Experimentally induced DNA hypomethylation only increased expression of thromboxane synthase in the neutrophil-like cell line, whereas tumor necrosis factor-α, a neutrophil product, increased its expression in cultured vascular smooth muscle cells. Our study suggests that epigenetic mechanisms and release of tumor necrosis factor-α by infiltrating neutrophils could contribute to the increased expression of thromboxane synthase in maternal systemic blood vessels, contributing to the hypertension and coagulation abnormalities associated with preeclampsia.

  13. Antioxidant Activity of the Essential Oil and its Major Terpenes of Satureja macrostema (Moc. and Sessé ex Benth.) Briq.

    Science.gov (United States)

    Torres-Martínez, Rafael; García-Rodríguez, Yolanda Magdalena; Ríos-Chávez, Patricia; Saavedra-Molina, Alfredo; López-Meza, Joel Edmundo; Ochoa-Zarzosa, Alejandra; Garciglia, Rafael Salgado

    2018-01-01

    The aim of this study was to investigate the in vitro antioxidant activity of Satureja macrostema (Moc. and Sessé ex Benth.) Briq. (Lamiaceae) essential oil, a Mexican medicinal plant known as nurite. Fresh aerial parts of S. macrostema plants cultivated in greenhouse for 3 months were subjected to hydrodistillation in a Clevenger apparatus to obtain essential oil. Volatile compounds were identified by gas chromatography (GC) and GC/mass spectrometry. Antioxidant effectiveness of essential oil and its major terpenes of S. macrostema was examined by three different radical scavenging methods: 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2'-azinobis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS), and total antioxidant capacity (TAC). The concentrations tested were 0.001, 0.01, 0.1, and 1 mg/mL. The major volatile compounds were caryophyllene, limonene, linalool, pulegone, menthone, and thymol. S. macrostema essential oil showed the highest free radical scavenging activity with DPPH and ABTS methods (53.10% and 92.12%, respectively) at 1 mg/mL and 98% with TAC method at 0.1 mg/mL. Thymol exerted the highest antioxidant capacity with 0.1 mg/mL, reaching 83.38%, 96.96%, and 98.57% by DPPH, ABTS, and TAC methods. Caryophyllene, limonene, linalool, pulegone, and menthone exhibited an antioxidant capacity essential oil of S. macrostema and thymol showed a free radical scavenging activity close to that of the synthetic butylated hydroxytoluene. The major volatile compounds of essential oil of Satureja macrostema were caryophyllene, limonene, linalool, pulegone, menthone and thymolThe essential oil of S. macrostema showed a high free radical scavengingThymol exerted the highest antioxidant capacity by DPPH, ABTS and TAC methods. Abbreviations used: GC: Gas Chromatography; DPPH: 2,2-diphenyl-1-picrylhydrazyl; ABTS: 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid; TAC: Total antioxidant capacity.

  14. Functional specificity of cardiolipin synthase revealed by the identification of a cardiolipin synthase CrCLS1 in Chlamydomonas reinhardtii

    Directory of Open Access Journals (Sweden)

    Chun-Hsien eHung

    2016-01-01

    Full Text Available Phosphatidylglycerol (PG and cardiolipin (CL are two essential classes of phospholipid in plants and algae. Phosphatidylglycerophosphate synthase (PGPS and cardiolipin synthase (CLS involved in the biosynthesis of PG and CL belong to CDP-alcohol phosphotransferase and share overall amino acid sequence homology. However, it remains elusive whether PGPS and CLS are functionally distinct in vivo. Here, we report identification of a gene encoding CLS in Chlamydomonas reinhardtii, CrCLS1, and its functional compatibility. Whereas CrCLS1 did not complement the growth phenotype of a PGPS mutant of Synechocystis sp. PCC 6803, it rescued the temperature-sensitive growth phenotype, growth profile with different carbon sources, phospholipid composition and enzyme activity of ∆crd1, a CLS mutant of Saccharomyces cerevisiae. These results suggest that CrCLS1 encodes a functional CLS of C. reinhardtii as the first identified algal CLS, whose enzyme function is distinct from that of PGPSs from C. reinhardtii. Comparison of CDP-alcohol phosphotransferase motif between PGPS and CLS among different species revealed a possible additional motif that might define the substrate specificity of these closely related enzymes.

  15. Geranylgeranyl diphosphate synthase from Scoparia dulcis and Croton sublyratus. Plastid localization and conversion to a farnesyl diphosphate synthase by mutagenesis.

    Science.gov (United States)

    Sitthithaworn, W; Kojima, N; Viroonchatapan, E; Suh, D Y; Iwanami, N; Hayashi, T; Noji, M; Saito, K; Niwa, Y; Sankawa, U

    2001-02-01

    cDNAs encoding geranylgeranyl diphosphate synthase (GGPPS) of two diterpene-producing plants, Scoparia dulcis and Croton sublyratus, have been isolated using the homology-based polymerase chain reaction (PCR) method. Both clones contained highly conserved aspartate-rich motifs (DDXX(XX)D) and their N-terminal residues exhibited the characteristics of chloroplast targeting sequence. When expressed in Escherichia coli, both the full-length and truncated proteins in which the putative targeting sequence was deleted catalyzed the condensation of farnesyl diphosphate and isopentenyl diphosphate to produce geranylgeranyl diphosphate (GGPP). The structural factors determining the product length in plant GGPPSs were investigated by constructing S. dulcis GGPPS mutants on the basis of sequence comparison with the first aspartate-rich motif (FARM) of plant farnesyl diphosphate synthase. The result indicated that in plant GGPPSs small amino acids, Met and Ser, at the fourth and fifth positions before FARM and Pro and Cys insertion in FARM play essential roles in determination of product length. Further, when a chimeric gene comprised of the putative transit peptide of the S. dulcis GGPPS gene and a green fluorescent protein was introduced into Arabidopsis leaves by particle gun bombardment, the chimeric protein was localized in chloroplasts, indicating that the cloned S. dulcis GGPPS is a chloroplast protein.

  16. Riboflavin accumulation and characterization of cDNAs encoding lumazine synthase and riboflavin synthase in bitter melon (Momordica charantia).

    Science.gov (United States)

    Tuan, Pham Anh; Kim, Jae Kwang; Lee, Sanghyun; Chae, Soo Cheon; Park, Sang Un

    2012-12-05

    Riboflavin (vitamin B2) is the universal precursor of the coenzymes flavin mononucleotide and flavin adenine dinucleotide--cofactors that are essential for the activity of a wide variety of metabolic enzymes in animals, plants, and microbes. Using the RACE PCR approach, cDNAs encoding lumazine synthase (McLS) and riboflavin synthase (McRS), which catalyze the last two steps in the riboflavin biosynthetic pathway, were cloned from bitter melon (Momordica charantia), a popular vegetable crop in Asia. Amino acid sequence alignments indicated that McLS and McRS share high sequence identity with other orthologous genes and carry an N-terminal extension, which is reported to be a plastid-targeting sequence. Organ expression analysis using quantitative real-time RT PCR showed that McLS and McRS were constitutively expressed in M. charantia, with the strongest expression levels observed during the last stage of fruit ripening (stage 6). This correlated with the highest level of riboflavin content, which was detected during ripening stage 6 by HPLC analysis. McLS and McRS were highly expressed in the young leaves and flowers, whereas roots exhibited the highest accumulation of riboflavin. The cloning and characterization of McLS and McRS from M. charantia may aid the metabolic engineering of vitamin B2 in crops.

  17. Application of a Colorimetric Assay to Identify Putative Ribofuranosylaminobenzene 5'-Phosphate Synthase Genes Expressed with Activity in Escherichia coli

    OpenAIRE

    Bechard, Matthew E.; Chhatwal, Sonya; Garcia, Rosemarie E.; Rasche, Madeline E.

    2003-01-01

    Tetrahydromethanopterin (H4MPT) is a tetrahydrofolate analog originally discovered in methanogenic archaea, but later found in other archaea and bacteria. The extent to which H4MPT occurs among living organisms is unknown. The key enzyme which distinguishes the biosynthetic pathways of H4MPT and tetrahydrofolate is ribofuranosylaminobenzene 5'-phosphate synthase (RFAP synthase). Given the importance of RFAP synthase in H4MPT biosynthesis, the identification of putative RFAP synthase genes and...

  18. Cloning and heterologous expression of a novel subgroup of class IV polyhydroxyalkanoate synthase genes from the genus Bacillus.

    Science.gov (United States)

    Mizuno, Kouhei; Kihara, Takahiro; Tsuge, Takeharu; Lundgren, Benjamin R; Sarwar, Zaara; Pinto, Atahualpa; Nomura, Christopher T

    2017-01-01

    Many microorganisms harbor genes necessary to synthesize biodegradable plastics known as polyhydroxyalkanoates (PHAs). We surveyed a genomic database and discovered a new cluster of class IV PHA synthase genes (phaRC). These genes are different in sequence and operon structure from any previously reported PHA synthase. The newly discovered PhaRC synthase was demonstrated to produce PHAs in recombinant Escherichia coli.

  19. Altering the expression of two chitin synthase genes differentially affects the growth and morphology of Aspergillus oryzae

    DEFF Research Database (Denmark)

    Müller, Christian; Hjort, C.M.; Hansen, K.

    2002-01-01

    In Aspergillus oryzae, one full-length chitin synthase (chsB) and fragments of two other chitin synthases (csmA and chsC) were identified. The deduced amino acid sequence of chsB was similar (87% identity) to chsB from Aspergillus nidulans, which encodes a class III chitin synthase. The sequence...

  20. Resistance to pathogens in terpene down-regulated orange fruits inversely correlates with the accumulation of D-limonene in peel oil glands.

    Science.gov (United States)

    Rodríguez, Ana; Shimada, Takehiko; Cervera, Magdalena; Redondo, Ana; Alquézar, Berta; Rodrigo, María Jesús; Zacarías, Lorenzo; Palou, Lluís; López, María M; Peña, Leandro

    2015-01-01

    Volatile organic compounds (VOCs) are secondary metabolites acting as a language for the communication of plants with the environment. In orange fruits, the monoterpene D-limonene accumulates at very high levels in oil glands from the peel. Drastic down-regulation of D-limonene synthase gene expression in the peel of transgenic oranges harboring a D-limonene synthase transgene in antisense (AS) configuration altered the monoterpene profile in oil glands, mainly resulting in reduced accumulation of D-limonene. This led to fruit resistance against Penicillium digitatum (Pd), Xanthomonas citri subsp. citri (Xcc) and other specialized pathogens. Here, we analyze resistance to pathogens in independent AS and empty vector (EV) lines, which have low, medium or high D-limonene concentrations and show that the level of resistance is inversely related to the accumulation of D-limonene in orange peels, thus explaining the need of high D-limonene accumulation in mature oranges in nature for the efficient attraction of specialized microorganism frugivores.

  1. Síntese de esteres terpenóides por via enzimática: influência do tamanho da cadeia alifática do ácido graxo e da estrutura do álcool de terpeno Synthesis of terpen esters by enzymatic route: influence of the fatty acid size chain and alcohol structure

    Directory of Open Access Journals (Sweden)

    Heizir F. CASTRO

    1997-12-01

    Full Text Available A especificidade de uma preparação comercial de lipase imobilizada, com relação a molécula ácida e alcoólica do substrato, foi estudada através da síntese de diversos ésteres de terpenóides. Na série de reações do citronelol e ácidos graxos com diferentes tamanho de cadeia alifática (C2 a C18, altas taxas de esterificação (95 a 98% foram alcançadas para ácidos contendo 4 ou mais carbonos. Numa segunda série de experimentos, diferentes álcoois terpenos foram esterificados com ácido butírico, sendo constatado uma influência marcante da estrutura do álcool de terpeno no desempenho desta preparação enzimática. Graus de esterificação maiores que 95% somente foram obtidos para os álcoois primários como citronelol, geraniol e nerol. Álcoois secundários (mentol e terciários (linalol não foram esterificados, sob as condições testadas.The selectivity of a commercial immobilized lipase preparation was tested in two set of esterification reactions. In the first group, synthesis were carried out with citronellol and different organic acids (C2 to C18. For this case, with the exception of acetic acid, the size of the carbon chain showed no significant alteration in the esterification rates. Acids containing four or more carbons, were considered to be excellent acyl donors, resulting in the esterification rates in the range of 95% to 98%. Alternatively, the esterification reactions were carried out with different terpen alcohols and butyric acid. The alcohol structure showed to have great influence on the performance of this enzyme preparation. Esterification degree over 95% were attained for primary alcohols such as citronellol, geraniol and nerol. Secondary (menthol and tertiary (linallol were not esterified under the tested conditions.

  2. Corn silk induces nitric oxide synthase in murine macrophages.

    Science.gov (United States)

    Kim, Kyung A; Choi, Sang Kyu; Choi, Hye Seon

    2004-12-31

    Corn silk has been purified as an anticoagulant previously and the active component is a polysaccharide with a molecular mass of 135 kDa. It activates murine macrophages to induce nitric oxide synthase (NOS) and generate substantial amounts of NO in time and dose-dependent manners. It was detectable first at 15 h after stimulation by corn silk, peaked at 24 h, and undetectable by 48 h. Induction of NOS is inhibited by pyrolidine dithiocarbamate (PDTC) and genistein, an inhibitor of nuclear factor kappa B (NF-kappaB) and tyrosine kinase, respectively, indicating that iNOS stimulated by corn silk is associated with tyrosine kinase and NF-kappaB signaling pathways. IkappaB-alpha degradation was detectible at 10 min, and the level was restored at 120 min after treatment of corn silk. Corn silk induced nuclear translocation of NF-kappaB by phosphorylation and degradation of IkappaB-alpha.

  3. CTP synthase forms the cytoophidium in human hepatocellular carcinoma.

    Science.gov (United States)

    Chang, Chia-Chun; Jeng, Yung-Ming; Peng, Min; Keppeke, Gerson Dierley; Sung, Li-Ying; Liu, Ji-Long

    2017-12-15

    CTP synthase (CTPS) can aggregate into an intracellular macrostructure, the cytoophidium, in various organisms including human cells. Previous studies have shown that assembly of human CTPS cytoophidia may be correlated with the cellular metabolic status, and is able to promote the activity of CTPS. A correlation between the cytoophidium and cancer metabolism has been proposed but not yet been revealed. In the current study we provide clear evidence of the presence of CTPS cytoophidia in various human cancers and some non-cancerous tissues. Moreover, among 203 tissue samples of hepatocellular carcinoma, 56 (28%) samples exhibited many cytoophidia, whereas no cytoophidia were detected in adjacent non-cancerous hepatocytes for all samples. Our findings suggest that the CTPS cytoophidium may participate in the adaptive metabolism of human hepatocellular carcinoma. Copyright © 2017. Published by Elsevier Inc.

  4. Reduced ceramide synthase 2 activity causes progressive myoclonic epilepsy

    DEFF Research Database (Denmark)

    Mosbech, Mai-Britt; Olsen, Anne S B; Neess, Ditte

    2014-01-01

    between genes involved in SL metabolism and epilepsy. METHODS: We used quantitative real-time PCR, Western blotting, and enzymatic assays to determine the mRNA, protein, and activity levels of ceramide synthase 2 (CERS2) in fiibroblasts isolated from parental control subjects and from a patient diagnosed...... with progressive myoclonic epilepsy (PME). Mass spectrometry and fluorescence microscopy were used to examine the effects of reduced CERS2 activity on cellular lipid composition and plasma membrane functions. RESULTS: We identify a novel 27 kb heterozygous deletion including the CERS2 gene in a proband diagnosed...... with PME. Compared to parental controls, levels of CERS2 mRNA, protein, and activity were reduced by ˜50% in fibroblasts isolated from this proband, resulting in significantly reduced levels of ceramides and sphingomyelins containing the very long-chain fatty acids C24:0 and C26:0. The change in SL...

  5. Sphingomyelin Synthase 1 Is Essential for Male Fertility in Mice.

    Directory of Open Access Journals (Sweden)

    Anke Wittmann

    Full Text Available Sphingolipids and the derived gangliosides have critical functions in spermatogenesis, thus mutations in genes involved in sphingolipid biogenesis are often associated with male infertility. We have generated a transgenic mouse line carrying an insertion in the sphingomyelin synthase gene Sms1, the enzyme which generates sphingomyelin species in the Golgi apparatus. We describe the spermatogenesis defect of Sms1-/- mice, which is characterized by sloughing of spermatocytes and spermatids, causing progressive infertility of male homozygotes. Lipid profiling revealed a reduction in several long chain unsaturated phosphatidylcholins, lysophosphatidylcholins and sphingolipids in the testes of mutants. Multi-Spectral Optoacoustic Tomography indicated blood-testis barrier dysfunction. A supplementary diet of the essential omega-3 docosahexaenoic acid and eicosapentaenoic acid diminished germ cell sloughing from the seminiferous epithelium and restored spermatogenesis and fertility in 50% of previously infertile mutants. Our findings indicate that SMS1 has a wider than anticipated role in testis polyunsaturated fatty acid homeostasis and for male fertility.

  6. Cellulose synthases: new insights from crystallography and modeling.

    Science.gov (United States)

    Slabaugh, Erin; Davis, Jonathan K; Haigler, Candace H; Yingling, Yaroslava G; Zimmer, Jochen

    2014-02-01

    Detailed information about the structure and biochemical mechanisms of cellulose synthase (CelS) proteins remained elusive until a complex containing the catalytic subunit (BcsA) of CelS from Rhodobacter sphaeroides was crystalized. Additionally, a 3D structure of most of the cytosolic domain of a plant CelS (GhCESA1 from cotton, Gossypium hirsutum) was produced by computational modeling. This predicted structure contributes to our understanding of how plant CelS proteins may be similar and different as compared with BcsA. In this review, we highlight how these structures impact our understanding of the synthesis of cellulose and other extracellular polysaccharides. We show how the structures can be used to generate hypotheses for experiments testing mechanisms of glucan synthesis and translocation in plant CelS. Copyright © 2013 Elsevier Ltd. All rights reserved.

  7. Two Cycloartenol Synthases for Phytosterol Biosynthesis in Polygala tenuifolia Willd.

    Science.gov (United States)

    Jin, Mei Lan; Lee, Woo Moon; Kim, Ok Tae

    2017-11-15

    Oxidosqualene cyclases (OSCs) are enzymes that play a key role in control of the biosynthesis of phytosterols and triterpene saponins. In order to uncover OSC genes from Polygala tenuifolia seedlings induced by methyl jasmonate (MeJA), RNA-sequencing analysis was performed using the Illumina sequencing platform. A total of 148,488,632 high-quality reads from two samples (control and the MeJA treated) were generated. We screened genes related to phytosterol and triterpene saponin biosynthesis and analyzed the transcriptional changes of differentially expressed unigene (DEUG) values calculated by fragments per kilobase million (FPKM). In our datasets, two full-length cDNAs of putative OSC genes, PtCAS1 , and PtCAS2 , were found, in addition to the PtBS (β-amyrin synthase) gene reported in our previous studies and the two cycloartenol synthase genes of P. tenuifolia . All genes were isolated and characterized in yeast cells. The functional expression of the two PtCAS genes in yeast cells showed that the genes all produce a cycloartenol as the sole product. When qRT-PCR analysis from different tissues was performed, the expressions of PtCAS1 and PtCAS2 were highest in flowers and roots, respectively. After MeJA treatment, the transcripts of PtCAS1 and PtCAS2 genes increased by 1.5- and 2-fold, respectively. Given these results, we discuss the potential roles of the two PtCAS genes in relation to triterpenoid biosynthesis.

  8. Two Cycloartenol Synthases for Phytosterol Biosynthesis in Polygala tenuifolia Willd

    Directory of Open Access Journals (Sweden)

    Mei Lan Jin

    2017-11-01

    Full Text Available Oxidosqualene cyclases (OSCs are enzymes that play a key role in control of the biosynthesis of phytosterols and triterpene saponins. In order to uncover OSC genes from Polygala tenuifolia seedlings induced by methyl jasmonate (MeJA, RNA-sequencing analysis was performed using the Illumina sequencing platform. A total of 148,488,632 high-quality reads from two samples (control and the MeJA treated were generated. We screened genes related to phytosterol and triterpene saponin biosynthesis and analyzed the transcriptional changes of differentially expressed unigene (DEUG values calculated by fragments per kilobase million (FPKM. In our datasets, two full-length cDNAs of putative OSC genes, PtCAS1, and PtCAS2, were found, in addition to the PtBS (β-amyrin synthase gene reported in our previous studies and the two cycloartenol synthase genes of P. tenuifolia. All genes were isolated and characterized in yeast cells. The functional expression of the two PtCAS genes in yeast cells showed that the genes all produce a cycloartenol as the sole product. When qRT-PCR analysis from different tissues was performed, the expressions of PtCAS1 and PtCAS2 were highest in flowers and roots, respectively. After MeJA treatment, the transcripts of PtCAS1 and PtCAS2 genes increased by 1.5- and 2-fold, respectively. Given these results, we discuss the potential roles of the two PtCAS genes in relation to triterpenoid biosynthesis.

  9. The evolution of function in strictosidine synthase-like proteins.

    Science.gov (United States)

    Hicks, Michael A; Barber, Alan E; Giddings, Lesley-Ann; Caldwell, Jenna; O'Connor, Sarah E; Babbitt, Patricia C

    2011-11-01

    The exponential growth of sequence data provides abundant information for the discovery of new enzyme reactions. Correctly annotating the functions of highly diverse proteins can be difficult, however, hindering use of this information. Global analysis of large superfamilies of related proteins is a powerful strategy for understanding the evolution of reactions by identifying catalytic commonalities and differences in reaction and substrate specificity, even when only a few members have been biochemically or structurally characterized. A comparison of >2500 sequences sharing the six-bladed β-propeller fold establishes sequence, structural, and functional links among the three subgroups of the functionally diverse N6P superfamily: the arylesterase-like and senescence marker protein-30/gluconolactonase/luciferin-regenerating enzyme-like (SGL) subgroups, representing enzymes that catalyze lactonase and related hydrolytic reactions, and the so-called strictosidine synthase-like (SSL) subgroup. Metal-coordinating residues were identified as broadly conserved in the active sites of all three subgroups except for a few proteins from the SSL subgroup, which have been experimentally determined to catalyze the quite different strictosidine synthase (SS) reaction, a metal-independent condensation reaction. Despite these differences, comparison of conserved catalytic features of the arylesterase-like and SGL enzymes with the SSs identified similar structural and mechanistic attributes between the hydrolytic reactions catalyzed by the former and the condensation reaction catalyzed by SS. The results also suggest that despite their annotations, the great majority of these >500 SSL sequences do not catalyze the SS reaction; rather, they likely catalyze hydrolytic reactions typical of the other two subgroups instead. This prediction was confirmed experimentally for one of these proteins. Copyright © 2011 Wiley-Liss, Inc.

  10. Intestinal nitric oxide synthase activity changes during experimental colon obstruction.

    Science.gov (United States)

    Palásthy, Zsolt; Kaszaki, József; Lázár, György; Nagy, Sándor; Boros, Mihály

    2006-08-01

    The experiments in this study were designed to follow the time course of nitric oxide (NO) synthesis in the large bowel during acute mechanical ileus. Occlusion of the mid-transverse colon was maintained for 420 min in anesthetized dogs. Strain-gauge transducers were used to analyze motility changes on the hepatic and lienal flexures, respectively. Constitutive NO synthase (cNOS) and inducible NOS (iNOS) activities were determined in tissue biopsies, and plasma nitrite/nitrate (NOx) level was measured in the portal blood. Following completion of the baseline studies, the animals were treated with either 7-nitroindazole (7-NI, selective neuronal NOS inhibitor), or N-nitro-L-arginine (NNA, non-selective NOS inhibitor). In the sham-operated group the cNOS activities differed significantly in the oral and aboral tissue samples (oral: 102.9; versus aboral: 62.1 fmol/mg protein/min). The obstruction elicited a significant increase in portal NOx and elevated tissue inducible NO synthase (iNOS) activity. NNA treatment decreased the motility index in both intestinal segments for 60 min, but 120 min later the motility index was significantly elevated (2.5-fold increase in the oral part, and 1.8-fold enhancement in the aboral segment, respectively). Treatment with 7-NI decreased the cNOS activity in the oral and aboral parts by approximately 40% and 70%, respectively, and suppressed the motility increase in the aboral colon segment. The motility of the colon was either significantly increased or decreased, depending on the type and selectivity of the NOS inhibitor compounds applied. NO of neuronal origin is a transmitter that stimulates peristaltic activity; but an increased iNOS/nNOS ratio significantly moderates the obstruction-induced motility increase.

  11. Glycogen Synthase in Sertoli Cells: More Than Glycogenesis?

    Science.gov (United States)

    Maldonado, Rodrigo; Mancilla, Héctor; Villarroel-Espíndola, Franz; Slebe, Felipe; Slebe, Juan Carlos; Méndez, Raúl; Guinovart, Joan J; Concha, Ilona I

    2016-11-01

    Sertoli cell metabolism actively maintains the nutritional needs of germ cells. It has been described that after glucose incorporation in Sertoli cells, less than 1% is converted to glycogen suggesting low levels of glycogen synthase activity. Phosphorylation of muscle glycogen synthase (MGS) at serine 640 (pS640MGS) decreases its activity, and this form of the enzyme was discovered as a non-ribosomal protein that modulates the translation of a subset of transcripts in HeLa cells. The aim of our study was to functionally characterize MGS in cultured Sertoli cells, as well as to explore this new feature related to RNA molecules. We detected MGS in the cytoplasm of Sertoli cells as well as in the nuclei. The activity rates of the enzyme were extremely low indicating that MGS is expressed but almost inactive. Protein targeting to glycogen (PTG) overexpression was performed to activate MGS by dephosphorylation. PTG induced glycogen synthesis massively, confirming that this enzyme is present but inactive. This finding correlates with high levels of pS640MGS, which were assayed by phosphatase treatment. To explore a putative new function for MGS in Sertoli cells, we performed RNA immunoprecipitation coupled to microarray studies. The results revealed that MGS co-immunoprecipitated with the several mRNAs and also rRNAs. These findings indicate that MGS is expressed Sertoli cells but in an inactive form, and also support a possibly novel feature of this metabolic enzyme associated with RNA-related molecules. J. Cell. Biochem. 117: 2597-2607, 2016. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

  12. Isolation and functional effects of monoclonal antibodies binding to thymidylate synthase.

    Science.gov (United States)

    Jastreboff, M M; Todd, M B; Malech, H L; Bertino, J R

    1985-01-29

    Monoclonal antibodies against electrophoretically pure thymidylate synthase from HeLa cells have been produced. Antibodies (M-TS-4 and M-TS-9) from hybridoma clones were shown by enzyme-linked immunoassay to recognize thymidylate synthase from a variety of human cell lines, but they did not bind to thymidylate synthase from mouse cell lines. The strongest binding of antibodies was observed to enzyme from HeLa cells. These two monoclonal antibodies bind simultaneously to different antigenic sites on thymidylate synthase purified from HeLa cells, as reflected by a high additivity index and results of cross-linked radioimmunoassay. Both monoclonal antibodies inhibit the activity of thymidylate synthase from human cell lines. The strongest inhibition was observed with thymidylate synthase from HeLa cells. Monoclonal antibody M-TS-9 (IgM subclass) decreased the rate of binding of [3H]FdUMP to thymidylate synthase in the presence of 5,10-methylenetetrahydrofolate while M-TS-4 (IgG1) did not change the rate of ternary complex formation. These data indicate that the antibodies recognize different epitopes on the enzyme molecule.

  13. Structure of the dimeric form of CTP synthase from Sulfolobus solfataricus

    DEFF Research Database (Denmark)

    Lauritsen, Iben; Willemoës, Martin; Jensen, Kaj Frank

    2011-01-01

    CTP synthase catalyzes the last committed step in de novo pyrimidine-nucleotide biosynthesis. Active CTP synthase is a tetrameric enzyme composed of a dimer of dimers. The tetramer is favoured in the presence of the substrate nucleotides ATP and UTP; when saturated with nucleotide, the tetramer...... completely dominates the oligomeric state of the enzyme. Furthermore, phosphorylation has been shown to regulate the oligomeric states of the enzymes from yeast and human. The crystal structure of a dimeric form of CTP synthase from Sulfolobus solfataricus has been determined at 2.5 Å resolution...

  14. Application of a Colorimetric Assay to Identify Putative Ribofuranosylaminobenzene 5'-Phosphate Synthase Genes Expressed with Activity in Escherichia coli

    Directory of Open Access Journals (Sweden)

    Bechard Matthew E.

    2003-01-01

    Full Text Available Tetrahydromethanopterin (H4MPT is a tetrahydrofolate analog originally discovered in methanogenic archaea, but later found in other archaea and bacteria. The extent to which H4MPT occurs among living organisms is unknown. The key enzyme which distinguishes the biosynthetic pathways of H4MPT and tetrahydrofolate is ribofuranosylaminobenzene 5'-phosphate synthase (RFAP synthase. Given the importance of RFAP synthase in H4MPT biosynthesis, the identification of putative RFAP synthase genes and measurement of RFAP synthase activity would provide an indication of the presence of H4MPT in untested microorganisms. Investigation of putative archaeal RFAP synthase genes has been hampered by the tendency of the resulting proteins to form inactive inclusion bodies in Escherichia coli. The current work describes a colorimetric assay for measuring RFAP synthase activity, and two modified procedures for expressing recombinant RFAP synthase genes to produce soluble, active enzyme. By lowering the incubation temperature during expression, RFAP synthase from Archaeoglobus fulgidus was produced in E. coli and purified to homogeneity. The production of active RFAP synthase from Methanothermobacter thermautotrophicus was achieved by coexpression of the gene MTH0830 with a molecular chaperone. This is the first direct biochemical identification of a methanogen gene that codes for an active RFAP synthase.

  15. Application of a Colorimetric Assay to Identify Putative Ribofuranosylaminobenzene 5'-Phosphate Synthase Genes Expressed with Activity in Escherichia coli.

    Science.gov (United States)

    Bechard, Matthew E.; Chhatwal, Sonya; Garcia, Rosemarie E.; Rasche, Madeline E.

    2003-01-01

    Tetrahydromethanopterin (H(4)MPT) is a tetrahydrofolate analog originally discovered in methanogenic archaea, but later found in other archaea and bacteria. The extent to which H(4)MPT occurs among living organisms is unknown. The key enzyme which distinguishes the biosynthetic pathways of H(4)MPT and tetrahydrofolate is ribofuranosylaminobenzene 5'-phosphate synthase (RFAP synthase). Given the importance of RFAP synthase in H(4)MPT biosynthesis, the identification of putative RFAP synthase genes and measurement of RFAP synthase activity would provide an indication of the presence of H(4)MPT in untested microorganisms. Investigation of putative archaeal RFAP synthase genes has been hampered by the tendency of the resulting proteins to form inactive inclusion bodies in Escherichia coli. The current work describes a colorimetric assay for measuring RFAP synthase activity, and two modified procedures for expressing recombinant RFAP synthase genes to produce soluble, active enzyme. By lowering the incubation temperature during expression, RFAP synthase from Archaeoglobus fulgidus was produced in E. coli and purified to homogeneity. The production of active RFAP synthase from Methanothermobacter thermautotrophicus was achieved by coexpression of the gene MTH0830 with a molecular chaperone. This is the first direct biochemical identification of a methanogen gene that codes for an active RFAP synthase.

  16. Effect of Transplanting Dates and Spacing on Yield Attributing Character, Productivity and Economics of Potato Cultivation Through True Potato Seed (TPS Technology

    Directory of Open Access Journals (Sweden)

    Debashish Sen

    2010-03-01

    Full Text Available A field experiment in split plot design was conducted during the rabi season of 2001-02 and 2002-03 at Potato Research and Seed Multiplication Farm, Anandapur,West Midnapore,West Bengal to study the effect of dates of transplanting and spacing on yield attributing character, productivity and economics of potato cultivation through true potato seed (TPS technology. The highest number of tubers per plant was recorded in early transplanted (December 3 crop, while, crop spacing did not produce any significant differences in recording tuber number per plant. Early established crop also produced significantly higher tuber weight per plant as compared to intermediate (December 11 and late (December 19 transplanted crops and widely spaced (60 x 15 cm crop recorded higher weight of tuber per plant as compared to the narrowly spaced crop. Early transplanting and wider spacing also showed its superiority in respect of recording harvest index of potato. As such, early transplanted and densely planted crops produced significantly higher yield of seedling tuber, marketable tuber and total tuber than their counterparts. Though cost of production of early transplanted and closely spaced crop was higher, but net profit and return per rupee investment was also higher in those cases.

  17. Targeted isolation, sequence assembly and characterization of two white spruce (Picea glauca BAC clones for terpenoid synthase and cytochrome P450 genes involved in conifer defence reveal insights into a conifer genome

    Directory of Open Access Journals (Sweden)

    Ritland Carol

    2009-08-01

    Full Text Available Abstract Background Conifers are a large group of gymnosperm trees which are separated from the angiosperms by more than 300 million years of independent evolution. Conifer genomes are extremely large and contain considerable amounts of repetitive DNA. Currently, conifer sequence resources exist predominantly as expressed sequence tags (ESTs and full-length (FLcDNAs. There is no genome sequence available for a conifer or any other gymnosperm. Conifer defence-related genes often group into large families with closely related members. The goals of this study are to assess the feasibility of targeted isolation and sequence assembly of conifer BAC clones containing specific genes from two large gene families, and to characterize large segments of genomic DNA sequence for the first time from a conifer. Results We used a PCR-based approach to identify BAC clones for two target genes, a terpene synthase (3-carene synthase; 3CAR and a cytochrome P450 (CYP720B4 from a non-arrayed genomic BAC library of white spruce (Picea glauca. Shotgun genomic fragments isolated from the BAC clones were sequenced to a depth of 15.6- and 16.0-fold coverage, respectively. Assembly and manual curation yielded sequence scaffolds of 172 kbp (3CAR and 94 kbp (CYP720B4 long. Inspection of the genomic sequences revealed the intron-exon structures, the putative promoter regions and putative cis-regulatory elements of these genes. Sequences related to transposable elements (TEs, high complexity repeats and simple repeats were prevalent and comprised approximately 40% of the sequenced genomic DNA. An in silico simulation of the effect of sequencing depth on the quality of the sequence assembly provides direction for future efforts of conifer genome sequencing. Conclusion We report the first targeted cloning, sequencing, assembly, and annotation of large segments of genomic DNA from a conifer. We demonstrate that genomic BAC clones for individual members of multi-member gene

  18. Targeted isolation, sequence assembly and characterization of two white spruce (Picea glauca) BAC clones for terpenoid synthase and cytochrome P450 genes involved in conifer defence reveal insights into a conifer genome.

    Science.gov (United States)

    Hamberger, Björn; Hall, Dawn; Yuen, Mack; Oddy, Claire; Hamberger, Britta; Keeling, Christopher I; Ritland, Carol; Ritland, Kermit; Bohlmann, Jörg

    2009-08-06

    Conifers are a large group of gymnosperm trees which are separated from the angiosperms by more than 300 million years of independent evolution. Conifer genomes are extremely large and contain considerable amounts of repetitive DNA. Currently, conifer sequence resources exist predominantly as expressed sequence tags (ESTs) and full-length (FL)cDNAs. There is no genome sequence available for a conifer or any other gymnosperm. Conifer defence-related genes often group into large families with closely related members. The goals of this study are to assess the feasibility of targeted isolation and sequence assembly of conifer BAC clones containing specific genes from two large gene families, and to characterize large segments of genomic DNA sequence for the first time from a conifer. We used a PCR-based approach to identify BAC clones for two target genes, a terpene synthase (3-carene synthase; 3CAR) and a cytochrome P450 (CYP720B4) from a non-arrayed genomic BAC library of white spruce (Picea glauca). Shotgun genomic fragments isolated from the BAC clones were sequenced to a depth of 15.6- and 16.0-fold coverage, respectively. Assembly and manual curation yielded sequence scaffolds of 172 kbp (3CAR) and 94 kbp (CYP720B4) long. Inspection of the genomic sequences revealed the intron-exon structures, the putative promoter regions and putative cis-regulatory elements of these genes. Sequences related to transposable elements (TEs), high complexity repeats and simple repeats were prevalent and comprised approximately 40% of the sequenced genomic DNA. An in silico simulation of the effect of sequencing depth on the quality of the sequence assembly provides direction for future efforts of conifer genome sequencing. We report the first targeted cloning, sequencing, assembly, and annotation of large segments of genomic DNA from a conifer. We demonstrate that genomic BAC clones for individual members of multi-member gene families can be isolated in a gene-specific fashion. The

  19. Eksperimentasi Pembelajaran Matematika dengan Model Kooperatif Tipe Nht dan Tps terhadap Prestasi Belajar dan Motivasi Berprestasi Siswa Ditinjau dari Kecerdasan Emosional Pokok Materi Persamaan Linear Satu Variabel pada Siswa SMP Negeri di Kota Surakarta

    OpenAIRE

    Aspriyani, Riski; Mardiyana, Mardiyana; Sari S, Dewi Retno

    2014-01-01

    This research was aimed to know: (1) which gave achievement and motivation to do better, students who were subjected to cooperative models of NHT, TPS, or Conventional and students with high, medium, or low emotional intelligence; (2) on each model of learning and at each category of emotional intelligence which provided achievement and motivation to do better. The population was all the students of VII grade of Junior High Schools in Surakarta year of 2013/2014 and obtained a sample of 26...

  20. The Sesquiterpenes(E-ß-Farnesene and (E-α-Bergamotene Quench Ozone but Fail to Protect the Wild Tobacco Nicotiana attenuata from Ozone, UVB, and Drought Stresses.

    Directory of Open Access Journals (Sweden)

    Evan C Palmer-Young

    Full Text Available Among the terpenes, isoprene (C5 and monoterpene hydrocarbons (C10 have been shown to ameliorate abiotic stress in a number of plant species via two proposed mechanisms: membrane stabilization and direct antioxidant effects. Sesquiterpene hydrocarbons (C15 not only share the structural properties thought to lend protective qualities to isoprene and monoterpene hydrocarbons, but also react rapidly with ozone, suggesting that sesquiterpenes may similarly enhance tolerance of abiotic stresses. To test whether sesquiterpenes protect plants against ozone, UVB light, or drought, we used transgenic lines of the wild tobacco Nicotiana attenuata. The transgenic plants expressed a maize terpene synthase gene (ZmTPS10 which produced a blend of (E-ß-farnesene and (E-α-bergamotene, or a point mutant of the same gene (ZmTPS10M which produced (E-ß-farnesene alone,. (E-ß-farnesene exerted a local, external, and transient ozone-quenching effect in ozone-fumigated chambers, but we found no evidence that enhanced sesquiterpene production by the plant inhibited oxidative damage, or maintained photosynthetic function or plant fitness under acute or chronic stress. Although the sesquiterpenes (E-ß-farnesene and (E-α-bergamotene might confer benefits under intermittent heat stress, which was not tested, any roles in relieving abiotic stress may be secondary to their previously demonstrated functions in biotic interactions.

  1. Molecular size estimation of plasma membrane β-glucan synthase from red beet root

    International Nuclear Information System (INIS)

    Sloan, M.E.; Eiberger, L.L.; Wasserman, B.P.

    1986-01-01

    Cellulose and cell wall β-D-glucans in higher plants are thought to be synthesized by the plasma membrane enzyme, β-glucan synthase. This enzyme has never been purified to homogeneity, hence its subunit composition is unknown. Partial purification of red beet root glucan synthase by glycerol density gradient centrifugation followed by SDS-PAGE yielded a highly enriched subunit of 68 kDa. Radiation inactivation of plasma membranes gave a molecular size the 450 kDa for the holoenzyme complex. This suggests that glucan synthase consists of 6 to 7 subunits and confirms electron microscope studies showing that glucan synthases exist as multi-subunit complexes embedded within the membrane

  2. SCREENING OF 6-PYRUVOYL-TETRAHYDROPTERIN SYNTHASE ACTIVITY DEFICIENCY AMONG HYPERP HENYLALANINEMIC PATIENTS

    Directory of Open Access Journals (Sweden)

    DURDI QUJEQ

    1999-10-01

    Full Text Available A deficiency of the phenylalanine hydroxylase activity or its cofactor tetrahydrobiopterin may"nlead to hyperphenylalamnemia and as a result, loss of IQ, poor school performance, and"nbehavior problems occurs. Deficiency in 6-pyruvoyl-tetrahydropterin synthase activity is the"nmajor cause of tetrahydrobiopterin deficient phenylketonuria. In this study, blood specimens"nfrom 165 healthy volunteers and 127 children with phenylketonuria were used to determine"nthe 6-pyruvoyl-tetrahydropterin synthase activity. It was found that the activity of 6-"npyruvoyl- tetrahydropterin synthase was decreased in comparison with control [23.46 +/-"n2.94, (mean +/- SD, mmol/ ml/h, n=I27 vs. 127.63 +/- 4.52, n=165, p<0.05]. Results of"nthis study indicate that examination of 6-pyruvoyl-tetrahydropterin synthase activity is helpful"nand may lead to the diagnosis cause of hyperphenylalaninemia.

  3. Mechanical Control of ATP Synthase Function: Activation Energy Difference between Tight and Loose Binding Sites

    KAUST Repository

    Beke-Somfai, Tamás; Lincoln, Per; Nordén, Bengt

    2010-01-01

    Despite exhaustive chemical and crystal structure studies, the mechanistic details of how FoF1-ATP synthase can convert mechanical energy to chemical, producing ATP, are still not fully understood. On the basis of quantum mechanical calculations

  4. Eukaryotic beta-alanine synthases are functionally related but have a high degree of structural diversity

    DEFF Research Database (Denmark)

    Gojkovic, Zoran; Sandrini, Michael; Piskur, Jure

    2001-01-01

    no pyrimidine catabolic pathway, it enabled growth on N-carbamyl- beta -alanine as the sole nitrogen source. The D. discoideum and D. melanogaster PYD3 gene products are similar to mammalian beta -alanine synthases. In contrast, the S. kluyveri protein is quite different from these and more similar to bacterial......beta -Alanine synthase (EC 3.5.1.6), which catalyzes the final step of pyrimidine catabolism, has only been characterized in mammals. A Saccharomyces kluyveri pyd3 mutant that is unable to grow on N-carbamy-beta -alanine as the sole nitrogen source and exhibits diminished beta -alanine synthase...... N- carbamyl amidohydrolases. All three beta -alanine synthases are to some degree related to various aspartate transcarbamylases, which catalyze the second step of the de novo pyrimidine biosynthetic pathway. PYD3 expression in yeast seems to be inducible by dihydrouracil and N...

  5. Citric acid production and citrate synthase genes in distinct strains of ...

    African Journals Online (AJOL)

    SAM

    2014-05-28

    May 28, 2014 ... synthase in lactic acid production by A. niger and with the ... A number of microorganisms, including both bacteria and fungi, possess the capacity ..... citric acid production by solid-state fermentation from cassava bagasse and ...

  6. Structure of an RNA dimer of a regulatory element from human thymidylate synthase mRNA

    OpenAIRE

    Dibrov, Sergey; McLean, Jaime; Hermann, Thomas

    2011-01-01

    An oligonucleotide representing a regulatory element of human thymidylate synthase mRNA has been crystallized as a dimer. The structure of the asymmetric dimer has been determined at 1.97 Å resolution.

  7. Improvement in the quality of hematopoietic prostaglandin D synthase crystals in a microgravity environment

    International Nuclear Information System (INIS)

    Tanaka, Hiroaki; Tsurumura, Toshiharu; Aritake, Kosuke; Furubayashi, Naoki; Takahashi, Sachiko; Yamanaka, Mari; Hirota, Erika; Sano, Satoshi; Sato, Masaru; Kobayashi, Tomoyuki; Tanaka, Tetsuo; Inaka, Koji; Urade, Yoshihiro

    2011-01-01

    Crystals of hematopoietic prostaglandin D synthase grown in microgravity show improved quality. Human hematopoietic prostaglandin synthase, one of the better therapeutic target enzymes for allergy and inflammation, was crystallized with 22 inhibitors and in three inhibitor-free conditions in microgravity. Most of the space-grown crystals showed better X-ray diffraction patterns than the terrestrially grown ones, indicating the advantage of a microgravity environment on protein crystallization, especially in the case of this protein

  8. Use of octaketide synthases to produce kermesic acid and flavokermesic acid

    DEFF Research Database (Denmark)

    2017-01-01

    A method for producing an octaketide derived aromatic compound of interest (e.g. carminic acid), wherein the method comprises (I): heterologous expression of a recombinantly introduced Type III polyketide synthase (PKS) gene encoding an octaketide synthase (OKS) to obtain non-reduced octaketide...... in vivo within the recombinant host cell and (II): converting in vivo the non-reduced octaketide of step (I) into a C14-C34 aromatic compound of interest (e.g. carminic acid)....

  9. Use of heterologous expressed polyketide synthase and small molecule foldases to make aromatic and cyclic compounds

    DEFF Research Database (Denmark)

    2016-01-01

    A method for producing individual or libraries of tri- to pentadecaketide-derived aromatic compounds of interest by heterologous expression of polyketide synthase and aromatase/cyclase in a recombinant host cell.......A method for producing individual or libraries of tri- to pentadecaketide-derived aromatic compounds of interest by heterologous expression of polyketide synthase and aromatase/cyclase in a recombinant host cell....

  10. Use of octaketide synthases to produce kermesic acid and flavokermesic acid

    DEFF Research Database (Denmark)

    2016-01-01

    A method for producing an octaketide derived aromatic compound of interest (e.g. carminic acid), wherein the method comprises (I): heterologous expression of a recombinantly introduced Type III polyketide synthase (PKS) gene encoding an octaketide synthase (OKS) to obtain non-reduced octaketide...... in vivo within the recombinant host cell and (II): converting in vivo the non-reduced octaketide of step (I) into a C14-C34 aromatic compound of interest (e.g. carminic acid)....

  11. The subcellular localization of yeast glycogen synthase is dependent upon glycogen content

    OpenAIRE

    Wilson, Wayne A.; Boyer, Michael P.; Davis, Keri D.; Burke, Michael; Roach, Peter J.

    2010-01-01

    The budding yeast, Saccharomyces cerevisiae, accumulates the storage polysaccharide glycogen in response to nutrient limitation. Glycogen synthase, the major form of which is encoded by the GSY2 gene, catalyzes the key regulated step in glycogen storage. Here, we utilize Gsy2p fusions to green fluorescent protein (GFP) to determine where glycogen synthase is located within cells. We demonstrate that the localization pattern of Gsy2-GFP depends upon the glycogen content of the cell. When glyco...

  12. The polyketide components of waxes and the Cer-cqu gene cluster encoding a novel polyketide synthase, the β-diketone synthase, DKS

    DEFF Research Database (Denmark)

    von Wettstein, Penny

    2017-01-01

    The primary function of the outermost, lipophilic layer of plant aerial surfaces, called the cuticle, is preventing non-stomatal water loss. Its exterior surface is often decorated with wax crystals, imparting a blue-grey color. Identification of the barley Cer-c, -q and -u genes forming the 101 kb...... Cer-cqu gene cluster encoding a novel polyketide synthase-the β-diketone synthase (DKS), a lipase/carboxyl transferase, and a P450 hydroxylase, respectively, establishes a new, major pathway for the synthesis of plant waxes. The major product is a β-diketone (14,16-hentriacontane) aliphatic that forms...

  13. Protein modelling of triterpene synthase genes from mangrove plants using Phyre2 and Swiss-model

    Science.gov (United States)

    Basyuni, M.; Wati, R.; Sulistiyono, N.; Hayati, R.; Sumardi; Oku, H.; Baba, S.; Sagami, H.

    2018-03-01

    Molecular cloning of five oxidosqualene cyclases (OSC) genes from Bruguiera gymnorrhiza, Kandelia candel, and Rhizophora stylosa had previously been cloned, characterized, and encoded mono and -multi triterpene synthases. The present study analyzed protein modelling of triterpene synthase genes from mangrove using Phyre2 and Swiss-model. The diversity was noted within protein modelling of triterpene synthases using Phyre2 from sequence identity (38-43%) and residue (696-703). RsM2 was distinguishable from others for template structure; it used lanosterol synthase as a template (PDB ID: w6j.1.A). By contrast, other genes used human lanosterol synthase (1w6k.1.A). The predicted bind sites were correlated with the product of triterpene synthase, the product of BgbAS was β-amyrin, while RsM1 contained a significant amount of β-amyrin. Similarly BgLUS and KcMS, both main products was lupeol, on the other hand, RsM2 with the outcome of taraxerol. Homology modelling revealed that 696 residues of BgbAS, BgLUS, RsM1, and RsM2 (91-92% of the amino acid sequence) had been modelled with 100% confidence by the single highest scoring template using Phyre2. This coverage was higher than Swiss-model (85-90%). The present study suggested that molecular cloning of triterpene genes provides useful tools for studying the protein modelling related regulation of isoprenoids biosynthesis in mangrove forests.

  14. Aspirin inhibits interleukin 1-induced prostaglandin H synthase expression in cultured endothelial cells

    International Nuclear Information System (INIS)

    Wu, K.K.; Sanduja, R.; Tsai, A.L.; Ferhanoglu, B.; Loose-Mitchell, D.S.

    1991-01-01

    Prostaglandin H (PGH) synthase is a key enzyme in the biosynthesis of prostaglandins, thromboxane, and prostacyclin. In cultured human umbilical vein endothelial cells, interleukin 1 (IL-1) is known to induce the synthesis of this enzyme, thereby raising the level of PGH synthase protein severalfold over the basal level. Pretreatment with aspirin at low concentrations inhibited more than 60% of the enzyme mass and also the cyclooxygenase activity in IL-1-induced cells with only minimal effects on the basal level of the synthase enzyme in cells without IL-1. Sodium salicylate exhibited a similar inhibitory action whereas indomethacin had no apparent effect. Similarly low levels of aspirin inhibited the increased L-[ 35 S]methionine incorporation i