WorldWideScience

Sample records for coprinus radians secretes

  1. The Coprophilous Mushroom Coprinus radians Secretes a Haloperoxidase That Catalyzes Aromatic Peroxygenation▿

    Science.gov (United States)

    Anh, Dau Hung; Ullrich, René; Benndorf, Dirk; Svatoś, Aleś; Muck, Alexander; Hofrichter, Martin

    2007-01-01

    Coprophilous and litter-decomposing species (26 strains) of the genus Coprinus were screened for peroxidase activities by using selective agar plate tests and complex media based on soybean meal. Two species, Coprinus radians and C. verticillatus, were found to produce peroxidases, which oxidized aryl alcohols to the corresponding aldehydes at pH 7 (a reaction that is typical for heme-thiolate haloperoxidases). The peroxidase of Coprinus radians was purified to homogeneity and characterized. Three fractions of the enzyme, CrP I, CrP II, and CrP III, with molecular masses of 43 to 45 kDa as well as isoelectric points between 3.8 and 4.2, were identified after purification by anion-exchange and size exclusion chromatography. The optimum pH of the major fraction (CrP II) for the oxidation of aryl alcohols was around 7, and an H2O2 concentration of 0.7 mM was most suitable regarding enzyme activity and stability. The apparent Km values for ABTS [2,2′-azinobis(3-ethylbenzthiazolinesulfonic acid)], 2,6-dimethoxyphenol, benzyl alcohol, veratryl alcohol, and H2O2 were 49, 342, 635, 88, and 1,201 μM, respectively. The N terminus of CrP II showed 29% and 19% sequence identity to Agrocybe aegerita peroxidase (AaP) and chloroperoxidase, respectively. The UV-visible spectrum of CrP II was highly similar to that of resting-state cytochrome P450 enzymes, with the Soret band at 422 nm and additional maxima at 359, 542, and 571 nm. The reduced carbon monoxide complex showed an absorption maximum at 446 nm, which is characteristic of heme-thiolate proteins. CrP brominated phenol to 2- and 4-bromophenols and selectively hydroxylated naphthalene to 1-naphthol. Hence, after AaP, CrP is the second extracellular haloperoxidase-peroxygenase described so far. The ability to extracellularly hydroxylate aromatic compounds seems to be the key catalytic property of CrP and may be of general significance for the biotransformation of poorly available aromatic substances, such as lignin, humus

  2. The coprophilous mushroom Coprinus radians secretes a haloperoxidase that catalyzes aromatic peroxygenation.

    Science.gov (United States)

    Anh, Dau Hung; Ullrich, René; Benndorf, Dirk; Svatos, Ales; Muck, Alexander; Hofrichter, Martin

    2007-09-01

    Coprophilous and litter-decomposing species (26 strains) of the genus Coprinus were screened for peroxidase activities by using selective agar plate tests and complex media based on soybean meal. Two species, Coprinus radians and C. verticillatus, were found to produce peroxidases, which oxidized aryl alcohols to the corresponding aldehydes at pH 7 (a reaction that is typical for heme-thiolate haloperoxidases). The peroxidase of Coprinus radians was purified to homogeneity and characterized. Three fractions of the enzyme, CrP I, CrP II, and CrP III, with molecular masses of 43 to 45 kDa as well as isoelectric points between 3.8 and 4.2, were identified after purification by anion-exchange and size exclusion chromatography. The optimum pH of the major fraction (CrP II) for the oxidation of aryl alcohols was around 7, and an H2O2 concentration of 0.7 mM was most suitable regarding enzyme activity and stability. The apparent Km values for ABTS [2,2'-azinobis(3-ethylbenzthiazolinesulfonic acid)], 2,6-dimethoxyphenol, benzyl alcohol, veratryl alcohol, and H2O2 were 49, 342, 635, 88, and 1,201 microM, respectively. The N terminus of CrP II showed 29% and 19% sequence identity to Agrocybe aegerita peroxidase (AaP) and chloroperoxidase, respectively. The UV-visible spectrum of CrP II was highly similar to that of resting-state cytochrome P450 enzymes, with the Soret band at 422 nm and additional maxima at 359, 542, and 571 nm. The reduced carbon monoxide complex showed an absorption maximum at 446 nm, which is characteristic of heme-thiolate proteins. CrP brominated phenol to 2- and 4-bromophenols and selectively hydroxylated naphthalene to 1-naphthol. Hence, after AaP, CrP is the second extracellular haloperoxidase-peroxygenase described so far. The ability to extracellularly hydroxylate aromatic compounds seems to be the key catalytic property of CrP and may be of general significance for the biotransformation of poorly available aromatic substances, such as lignin

  3. Guanacastane-type diterpenoids from Coprinus radians.

    Science.gov (United States)

    Ou, Yi-xin; Li, Yao-yao; Qian, Xiao-ming; Shen, Yue-mao

    2012-06-01

    Thirteen diterpenoids, named radianspenes A-M (1-13), including three lactams radianspenes J (10), K (11) and L (12) and one dimer radianspene M (13), were isolated from fermentation products of the higher fungal strain Coprinus radians M65. All these compounds possessing guanacastane skeleton were evaluated for antitumor activity using MDA-MB-435 cell line. Radianspene C exhibited inhibitory activity with IC(50) of 0.91 μM. Copyright © 2012 Elsevier Ltd. All rights reserved.

  4. Experiencing Radians.

    Science.gov (United States)

    Eggleton, Patrick J.

    1999-01-01

    Presents an activity that uses an inexpensive paper plate and a short length of adding machine tape to help students experience the concept of radian measure. Focuses on radian measurement and connects many geometric concepts regarding the circle. (ASK)

  5. Walking a Radian

    Science.gov (United States)

    Touval, Ayana

    2009-01-01

    The kinematics teaching strategy is a teaching method that stimulates kinesthetic intelligence and thus offers students an unconventional approach for exploring mathematical ideas through movement. This article describes how to use the kinesthetic approach to introduce radian measure. The article includes detailed descriptions of easy-to-use…

  6. Studies in Coprinus IV — Coprinus section coprinus. Subdivision and revision of subsection Alachuani

    OpenAIRE

    Uljé, C.B.; Noordeloos, M.E.

    1997-01-01

    Coprinus section Coprinus is defined and delimited to comprise four subsections: Atramentarii, Coprinus, Lanatuli and Alachuani. A key to the subsections is given as well as a key to the species of subsection Alachuani known from the Netherlands or to be expected in the Netherlands on account of records from neighbouring countries. Three new species, Coprinus epichloeus, Coprinus fluvialis and Coprinus sclerotiorum are described as well as a new variety of C. urticicola: var. salicicola. In a...

  7. The Radian--That Troublesome Unit.

    Science.gov (United States)

    American Association of Physics Teachers

    1993-01-01

    Defines and discusses the proper use of radians in physics equations and problem solving. The article suggests that many physics instructors inadequately explain how and when to use radians to their students. (MVL)

  8. Radian: Visual Exploration of Traceroutes.

    Science.gov (United States)

    Candela, Massimo; Di Bartolomeo, Marco; Di Battista, Giuseppe; Squarcella, Claudio

    2017-06-19

    Several projects deploy probes in the Internet. Probes are systems that continuously perform traceroutes and other networking measurements (e.g. ping) towards selected targets. Measurements can be stored and analyzed to gain knowledge on several aspects of the Internet, but making sense of such data requires suitable methods and tools for exploration and visualization. We present Radian, a tool that allows to visualize traceroute paths at different levels of detail and to animate their evolution during a selected time interval. We also describe extensive tests of the tool using traceroutes performed by RIPE Atlas Internet probes.

  9. Studies in Coprinus V — Coprinus section Coprinus. Revision of subsection Lanatuli Sing

    NARCIS (Netherlands)

    Uljé, C.B.; Noordeloos, M.E.

    1999-01-01

    A key is given to the species of subsection Lanatuli known from the Netherlands or to be expected in the Netherlands on account of records from neighbouring countries. For a key to the subsections in Coprinus section Coprinus see Uljé & Noordel., Persoonia 16 (1997) 267. Coprinus bicornis and C.

  10. Studies in Coprinus IV — Coprinus section coprinus. Subdivision and revision of subsection Alachuani

    NARCIS (Netherlands)

    Uljé, C.B.; Noordeloos, M.E.

    1997-01-01

    Coprinus section Coprinus is defined and delimited to comprise four subsections: Atramentarii, Coprinus, Lanatuli and Alachuani. A key to the subsections is given as well as a key to the species of subsection Alachuani known from the Netherlands or to be expected in the Netherlands on account of

  11. Studies in Coprinus V — Coprinus section Coprinus. Revision of subsection Lanatuli Sing

    OpenAIRE

    Uljé, C.B.; Noordeloos, M.E.

    1999-01-01

    A key is given to the species of subsection Lanatuli known from the Netherlands or to be expected in the Netherlands on account of records from neighbouring countries. For a key to the subsections in Coprinus section Coprinus see Uljé & Noordel., Persoonia 16 (1997) 267. Coprinus bicornis and C. spelaiophilus are described as new species. In addition the following species are fully described: C. ammophilae, C. calosporus, C. cinereus, C. erythrocephalus, C. geesterani, C. jonesii, C. krieglst...

  12. Using Technology To Introduce Radian Measure.

    Science.gov (United States)

    Klein, Raymond J.; Hamilton, Ilene

    1997-01-01

    Describes a method of effectively introducing the concept of radian measure that uses Cabri Geometry II software to construct a circle of arbitrary radius and to measure that radius. The goal is to determine how many of these radii fit around a circle. (DDR)

  13. First report of Coprinus spadiceisporus in Europe

    NARCIS (Netherlands)

    Uljé, C.B.; Gennari, A.; Doveri, F.; Cacialli, G.; Caroti, V.

    1998-01-01

    The first record of Coprinus spadiceisporus Van De Bogart in Europe is described. A study of the type and that of Coprinus roseistipitatus Van De Bogart revealed that both are conspecific, and accordingly the latter name is considered synonymous.

  14. Type studies in Coprinus subsection Lanatuli

    NARCIS (Netherlands)

    Uljé, C.B.; Noordeloos, M.E.

    2001-01-01

    As a prelude to a monograph of the genus Coprinus, types were studied of a number of species said to belong to Coprinus subsection Lanatuli (Coprinus alnivorus. C. alutaceivelatus, C. ammophilae, C. arachnoideus, C. asterophoroides, C. brunneistragulatus, C. bubalinus, C. citrinovelatus, C.

  15. Pre-Service Mathematics Teachers' Concept Images of Radian

    Science.gov (United States)

    Akkoc, Hatice

    2008-01-01

    This study investigates pre-service mathematics teachers' concept images of radian and possible sources of such images. A multiple-case study was conducted for this study. Forty-two pre-service mathematics teachers completed a questionnaire, which aims to assess their understanding of radian. Six of them were selected for individual interviews on…

  16. Additional studies in Coprinus subsection Glabri

    NARCIS (Netherlands)

    Uljé, C.B.; Bender, H.

    1997-01-01

    First Coprinus lilatinctus, belonging to subsect. Glabri, is described as new. Secondly, nomenclatural reasons are given for Coprinus nudiceps P.D. Orton to be replaced by the older name C. schroeteri P. Karst. Type studies of both taxa are given and their synonymy is discussed. To facilitate

  17. A new species in Coprinus subsection Setulosi

    NARCIS (Netherlands)

    Uljé, C.B.; Verbeken, A.

    2002-01-01

    Coprinus canistri spec. nov. is proposed. It belongs to the subsection Setulosi because of the presence of pileo- and caulocystidia. A comparison is given with C. subimpatiens and C. congregatus, on account of similar microscopical characters.

  18. Some new species of Coprinus from the Netherlands

    NARCIS (Netherlands)

    Uljé, C.B.; Bas, C.

    1993-01-01

    Four new species of Coprinus are illustrated and described as C. calosporus and C. ochraceolanatus in sect. Coprinus subsect. Lanatuli (Fr.) Sing. and C. goudensis and C. piepenbroekii in sect. Coprinus subsect. Alachuani Sing. (= Impexi s. Kühn. & Rom., 1953).

  19. Meiotic cytogenetics in Coprinus cinereus.

    Science.gov (United States)

    Zolan, Miriam E; Pukkila, Patricia J

    2009-01-01

    The basidiomycete fungus Coprinus cinereus has naturally synchronous meiosis and is amenable to analysis using an array of well-developed genetic and molecular tools. In this chapter, we explain in detail the two methods most commonly employed for C. cinereus, staining of intact gill segments and chromosome spreads, with an example of the application of each. We describe iron-hematoxylin staining of intact gill segments for the brightfield examination of meiotic progression, and the use of surface spreads and fluorescence in situ hybridization (FISH) to investigate meiotic chromosome pairing. Gill segments can alternatively be stained with DAPI for the determination of meiotic stage, or propidium iodide for the quantitation of nuclear DNA content, and the chromosome fixation and spreading techniques used for FISH are also suitable for immunolocalization studies of chromosomal proteins.

  20. [Expression and characterization of Coprinus cinereus peroxidase].

    Science.gov (United States)

    Dong, Bingxue; Niu, Qiuhong; Zhang, Wei; Geng, Sanchun; Li, Peng; Yuan, Wanli; Gong, Yifeng; Liang, Kewei

    2015-03-04

    The aim of our study is to express Coprinus cinereus peroxidase (CIP) in Pichia Pastori efficiently. We synthesized CIP gene with P. pastori codon bias by our Gene Synthesis and site-specific mutagenesis platform, using DNAWorks 3.1 program to design and optimize primers. Then, we sequenced the PCR products, inserted the correct gene into expression vector pPICZαA and transformed the linearized pPICZαA-Cip DNA into P. pastori GS115. We integrated CIP gene into the genome of P. pastori, using the α-mating factor from Sacchoramyces cerevisiae as signal peptide to direct the secretion of the recombinant protein. To obtain transformants with high CIP activity, we checked transformants by nested PCR and stained 82 positive ones on YPD agar plate with 1000 mg/L Zeocin. Then, we got 6 transforments with high resistance to Zeocin and expressed them in small scale; the one exhibiting the highest activity was chosen as engineered strain and named CIP/Gs115. We purified CIP from culture medium after induction with ethanol, the maximum activity reached 487.5 U/mL on the 4th day. The purified CIP exhibited maximal activity at pH 5.0 and 25 degrees C with ABTS as substrate. The enzyme had 61.5% of the maximal activity at 45 degrees C and was stable below 40 degrees C. However, the stability was drastically reduced above 45 degrees C. The recombinant CIP remained stable between pH 4.5 and 6.5. We studied the substrate specificity on different substrates with the purified enzyme, and the optimal substrates were in the order of ABTS > 2, 6-Dimethoxyphenol > guaiacol > 2, 4-Dichlorophenol > phenol. The highly secretory expression of CIP and high special activity lay the good foundation for it' s industrial applications in waste water treatment, decolouration of dyestuffs.

  1. Studies in Coprinus III — Coprinus section Veliformes. Subdivision and revision of subsection Nivei emend

    NARCIS (Netherlands)

    Uljé, C.B.; Noordeloos, M.E.

    1993-01-01

    Coprinus section Veliformes is defined and delimited to comprise four subsections: subsection Micacei, subsection Domestici, subsection Nivei, and subsection Narcotici, subsection nov. A key to the subsections is given. Subsection Nivei is emended, including also most taxa of subsection Flocculosi

  2. Introducing Radian: A Community-curated Hub for All Things Data

    OpenAIRE

    Mannheimer, Sara; Champieux, Robin; Young, Scott W. H.; Robinson, Danielle; Banta, Ryer

    2017-01-01

    Presentation given at the Force 2017 conference in Berlin, Germany on Radian, a community effort to increase the discovery and reuse of data resources. Use Radian to discover ideas and share resources about data science, visualization, data management, data policy, and applications of data for the public good.

  3. [Fourier transform infrared spectroscopic study of Coprinus comatus].

    Science.gov (United States)

    Zhou, Ji-Guo; Liu, Gang; Liu, Jian-Hong; Song, Ding-Shan; Shi, Yong-Ming

    2008-02-01

    In the paper, Fourier transform infrared (FTIR) spectroscopy was used to study the fruiting body of wild growing and cultivated Coprinus comatus and the cap of the mushroom before and after deliquescing into inky liquid. The results show that the infrared spectra of Coprinus comatus are mainly composed of the absorption bands of protein and polysaccharide. Remarkable differences are observed in the absorption bands of polysaccharide between the spectra of the wild growing and cultivated mushrooms, by which the wild growing and cultivated Coprinus comatus can be discriminated. It was also found that the absorption bands of polysaccharide become weaker obviously in the black-inky samples of deliquescing cap compared with the un-deliquescing cap, indicating that the polysaccharides in the cap of Coprinus comatus are transformed during the process of cap deliquescing into a black, inky liquid. The spectral results can offer useful information for a further study of Coprinus comatus.

  4. Optimization of extracellular fungal peroxidase production by 2 Coprinus species.

    Science.gov (United States)

    Ikehata, Keisuke; Pickard, Michael A; Buchanan, Ian D; Smith, Daniel W

    2004-12-01

    Optimum culture conditions for the batch production of extracellular peroxidase by Coprinus cinereus UAMH 4103 and Coprinus sp. UAMH 10067 were explored using 2 statistical experimental designs, including 2-level, 7-factor fractional factorial design and 2-factor central composite design. Of the 7 factors examined in the screening study, the concentrations of carbon (glucose) and nitrogen (peptone or casitone) sources showed significant effects on the peroxidase production by Coprinus sp. UAMH 10067. The optimum glucose and peptone concentrations were determined as 2.7% and 0.8% for Coprinus sp. UAMH 10067, and 2.9% and 1.4% for C. cinereus UAMH 4103, respectively. Under the optimized culture condition the maximum peroxidase activity achieved in this study was 34.5 U x mL(-1) for Coprinus sp. UAMH 10067 and 68.0 U x mL(-1) for C. cinereus UAMH 4103, more than 2-fold higher than the results of previous studies.

  5. Type studies in the genus Coprinus (Coprinaceae, Agaricales) Coprinus xerophilus a new record in Europe

    NARCIS (Netherlands)

    Moreno, G.; Heykoop, M.

    1998-01-01

    In the present study a comparison of the holotypes of Coprinus calyptratus, C. vosoustii, C. asterophorus and C. asterophoroides has been carried out. As a conclusion of the latter we can state that all of them are conspecific, the correct name of this taxon being C. calyptratus. On the other hand

  6. New bioactive natural products from Coprinus micaceus.

    Science.gov (United States)

    Zahid, Shamsulhaq; Udenigwe, Chibuike C; Ata, Athar; Eze, Michael O; Segstro, Edward P; Holloway, Paul

    2006-12-01

    Chemical studies on the crude methanolic extract of Coprinus micaceus, collected from the Canadian Prairie region, have resulted in the isolation of two new natural products, micaceol (1), a sterol and (Z,Z)-4-oxo-2,5-hetpadienedioic acid (2). Compound 2 was isolated for the first time as a natural product. Structures of these new compounds were established with the aid of extensive NMR spectroscopic studies. Compound 1 has shown anti-bacterial activity against Corynebacterium xerosis and Staphylococcus aureus, whereas compound 2 exhibited glutathione S-transferase inhibition.

  7. Inheritance of DNA methylation in Coprinus cinereus.

    Science.gov (United States)

    Zolan, M E; Pukkila, P J

    1986-01-01

    We examined the inheritance of 5-methylcytosine residues at a centromere-linked locus in the basidiomycete Coprinus cinereus. Although methylated and unmethylated tracts were inherited both mitotically and meiotically the lengths of these tracts were variable. This variation was not confined to any one phase of the life cycle of the organism, and it usually involved the simultaneous de novo methylation of at least four HpaII-MspI sites. We also found that the higher levels of methylation at this locus were transmitted through meiosis, regardless of the level of methylation of the homologous chromosome. Images PMID:3785146

  8. Tolerance of two Bifora radians bieb populations to ALS inhibitors in winter wheat

    DEFF Research Database (Denmark)

    Mennan, Husrev; Streibig, Jens Carl; Ngouajio, Mathieu

    2012-01-01

    allegedly sensitive population was estimated at the ED50 and ED90 response levels. The recommended rates of herbicides controlled 90% of the weed (ED90) in the sensitive population at the early stage of B. radians development, but not in the tolerant population. The relative potencies (EDx......BACKGROUND:Bifora radians, an annual weed in winter wheat, is distributed mainly in the Mediterranean area, Asia Minor and the Caucasus. It infests winter-sown crops of the Central Anatolia and Middle Black Sea regions of Turkey. Field experiments in heavily B. radians-infested fields were...... conducted over 3 years in Samsun, Turkey, to determine the response of B. radians to ALS-inhibiting herbicides, because growers had complained of a decrease in herbicide effect. RESULTS: The efficacy of ALS inhibitors on a putatively tolerant population sprayed annually with ALS inhibitors and an adjacent...

  9. Secretory production of Aspergillus oryzae xylanase XynF1, xynF1 cDNA product, in the basidiomycete Coprinus cinereus.

    Science.gov (United States)

    Kikuchi, M; Kitamoto, N; Shishido, K

    2004-02-01

    The signal peptide of Aspergillus oryzae endo-(1,4)-beta-xylanase XynF1 contains a C-terminal serine-arginine that directs efficient secretion of the enzyme into the culture medium. In the basidiomycete Coprinus cinereus, however, there is little secretion of XynF1 into the culture medium. Modification of the C-terminal sequence of the signal peptide to lysine-arginine resulted in efficient secretion of C. cinereus XynF1, suggesting the presence of a KEX2-like protease in this fungus.

  10. DNA-mediated transformation of the basidiomycete Coprinus cinereus.

    Science.gov (United States)

    Binninger, D M; Skrzynia, C; Pukkila, P J; Casselton, L A

    1987-01-01

    We have developed a simple and efficient transformation system for the agaric fungus, Coprinus cinereus. Protoplasts were prepared from asexual spores that harbor one or two mutations in the structural gene for tryptophan synthetase. The protoplasts can be stably transformed using the cloned Coprinus gene at a frequency of 1 in 10(4) viable protoplasts. A variety of molecular events accompanies the formation of stable transformants, including insertion of the transforming DNA at the homologous locus. The transforming DNA is stable through cell division, mating, fruiting body formation, and meiosis. Images Fig. 2. Fig. 3. Fig. 4. PMID:3595558

  11. DNA polymerase of a basidiomycete fungus, Coprinus cinereus.

    Science.gov (United States)

    Lowe, D; Lewis, D

    1981-01-01

    DNA polymerase activity was studied in Coprinus cinereus, a basidiomycete fungus. Only one from of the enzyme could be demonstrated, whether by affinity or ion-exchange chromatography; this enzyme had a molecular weight of 185000 on Sephadex G-200, and was inhibited by mercaptoethanol. Coprinus, a representative of the most advanced type of the filamentous fungi, resembles other eukaryotic micro-organisms in its lack of a mammalian beta-type DNA polymerase. The properties of the polymerase are compared with those of two other fungi, and found to resemble most closely the yeast polymerase A in Mg2+ requirements and template preference. PMID:7197920

  12. A Conceptual Analysis of the Knowledge of Prospective Mathematics Teachers about Degree and Radian

    Science.gov (United States)

    Tuna, Abdulkadir

    2013-01-01

    This study examined the knowledge levels of prospective mathematics teachers about the concepts of degree and radian, which are among the angle measuring units that constitute the basis of trigonometry, and the relationships between those concepts. The study group consisted of 93 prospective mathematics teachers attending a state university in…

  13. Solar heating and cooling demonstration project at Radian Corporation, Austin, Texas

    Science.gov (United States)

    1980-01-01

    The solar heating and cooling system located at the Radian Corporation, Austin, Texas, is discussed. A technical description of the solar system is presented. The costs of the major components and the cost of installing the system are described. Flow diagrams and photographs of the solar system are provided.

  14. Coprinus Idae Uljé (Coprinaceae, Agaricales) found in Spain

    NARCIS (Netherlands)

    Esteve-Raventós, F.; Cruz, de la M.

    1997-01-01

    A new record of Coprinus idae Uljé, this time collected from Spain, is briefly described, and some taxonomic and ecologic observations are made. Up to now, this species has been reported from the Netherlands, only twice from the type-locality.

  15. A new species of Coprinus from the Netherlands

    NARCIS (Netherlands)

    Ujlé, C.B.

    1992-01-01

    Several times a small, unknown species of Coprinus has been found on paths covered with wood-chips and on composted vegetable refuse. The basidiocarps of this species look like miniature fruit-bodies of C. lagopus. As in literature no suitable name could be found, it is described here as new. I

  16. Four new species of Coprinus from the Netherlands

    NARCIS (Netherlands)

    Uljé, C.B.

    1988-01-01

    Three species of Coprinus belonging to the C. niveus-C. cortinatus group are newly described, viz. C. bellulus, C. candidates, and C. iocularis. For purposes of comparison a description of C. cortinatus is given also. In addition C. singularis, belonging to subsection Setulosi, is introduced as a

  17. A new cyclic dipeptide from cultures of Coprinus plicatilis.

    Science.gov (United States)

    Zhao, Jiang-Yuan; Ding, Jian-Hai; Li, Zheng-Hui; Feng, Tao; Zhang, Hong-Bin; Liu, Ji-Kai

    2017-09-01

    Three cyclic dipeptides (1-3), including one new compound (1) were isolated from cultures of the basidiomycetes Coprinus plicatilis. Their structures were elucidated by spectroscopic methods, including extensive 2D NMR techniques. At the same time, all compounds were tested for their cytotoxicities against five human cancer cell lines.

  18. Cultivation of Tanzanian Coprinus cinereus (sisal compost mushroom)

    African Journals Online (AJOL)

    Coprinus cinereus is a Tanzanian wild edible mushroom whose cultivation in the laboratory was performed in solid-state fermentation bioreactors using sisal decortication wastes namely; sisal dusts, sisal fibres and sisal leaves as basal substrates supplemented with chicken manure at various rates. Bioreactors containing ...

  19. Extracellular peroxidase production by Coprinus species from urea-treated soil.

    Science.gov (United States)

    Ikehata, Keisuke; Buchanan, Ian D; Smith, Daniel W

    2004-01-01

    Thirteen strains of inky-cap mushroom Coprinus species were evaluated for the production of extracellular peroxidase. The liquid fermentation was carried out in shake flasks containing 1% glucose, 0.5% peptone, 0.3% yeast extract, and 0.3% malt extract broth at 25 degrees C. Peroxidase activity was detected in the liquid culture of several Coprinus species, including C. echinosporus NBRC 30630; C. macrocephalus NBRC 30117; Coprinus spp. UAMH 10065, UAMH 10066, UAMH 10067, and 074, after 10 days of growth. Peroxidase production by Coprinus sp. UAMH 10067, a Coprinus species isolated from urea-treated soil, was comparable to that of C. cinereus and reached 15 U.mL(-1) after 10 days. In addition, the peroxidase from Coprinus sp. UAMH 10067 was apparently more thermally stable than the enzyme produced by C. cinereus.

  20. Isolation and biological activity of triglycerides of the fermented mushroom of Coprinus Comatus

    OpenAIRE

    Ren Jun; Shi Jin-Li; Han Chun-Chao; Liu Zhen-Quan; Guo Jian-You

    2012-01-01

    Abstract Background Although many physiological functions of Coprinus comatus have been reported, there has been no report on the antinociceptive activity of Coprinus comatus. Therefore, the objective of the present study is to demonstrate the production, isolation, and biological properties of triglycerides (TFC) of the fermented mushroom of Coprinus comatus. Methods The effects of TFC on cytokines levels, total antioxidant activity, antinociceptive effects in vivo, LD50 and tactile hyperalg...

  1. Controllable optical phase shift over one radian from a single isolated atom.

    Science.gov (United States)

    Jechow, A; Norton, B G; Händel, S; Blūms, V; Streed, E W; Kielpinski, D

    2013-03-15

    Fundamental optics such as lenses and prisms work by applying phase shifts of several radians to incoming light, and rapid control of such phase shifts is crucial to telecommunications. However, large, controllable optical phase shifts have remained elusive for isolated quantum systems. We have used a single trapped atomic ion to induce and measure a large optical phase shift of 1.3±0.1 radians in light scattered by the atom. Spatial interferometry between the scattered light and unscattered illumination light enables us to isolate the phase shift in the scattered component. The phase shift achieves the maximum value allowed by atomic theory over the accessible range of laser frequencies, pointing out new opportunities in microscopy and nanophotonics. Single-atom phase shifts of this magnitude open up new quantum information protocols, in particular long-range quantum phase-shift-keying cryptography.

  2. Genetic identification of bucktooth parrotfish Sparisoma radians (Valenciennes, 1840 (Labridae, Scarinae by chromosomal and molecular markers

    Directory of Open Access Journals (Sweden)

    Fabilene Gomes Paim

    2014-12-01

    Full Text Available Parrotfishes (Labridae, Scarinae comprise a large marine fish group of difficult identification, particularly during juvenile phase when the typical morphology and coloration of adults are absent. Therefore, the goal of this study was to test cytogenetic markers and DNA barcoding in the identification of bucktooth parrtotfish Sparisoma radians from the northeastern coast of Brazil. Sequencing of cytochrome c oxidase subunit I (COI confirmed all studied samples as S. radians, and all showed high similarity (99-100% with Caribbean populations. The karyotype of this species was divergent from most marine Perciformes, being composed of 2n = 46 chromosomes. These consisted of a large number of metacentric and submetacentric pairs with small amounts of heterochromatin and GC-rich single nucleolar organizer regions (NORs not syntenic to 5S rDNA clusters. These are the first data about DNA barcoding in parrotfish from the Brazilian province and the first refined chromosomal analysis in Scarinae, providing useful data to a reliable genetic identification of S. radians.

  3. Studies in Coprinus—I. Subsections Auricomi and Glabri of Coprinus section Pseudocoprinus

    NARCIS (Netherlands)

    Uljé, C.B.; Bas, C.

    1988-01-01

    A key is given to the Netherlands’ species of subsect. Auricomi Sing. and Glabri J. Lange of Coprinus sect. Pseudocoprinus (Kühn.) Orton & Watling. All species concerned are concisely described and amply discussed. Coprinus plicatilis var. microsporus Kühn. is raised to species level as C. kuehneri

  4. The co-effect of vanadium and fermented mushroom of Coprinus comatus on glycaemic metabolism.

    Science.gov (United States)

    Zhou, Guangtian; Han, Chunchao

    2008-07-01

    The effect of fermented mushroom of Coprinus comatus rich in vanadium (CCRV) on glycaemic metabolism was studied in this paper. Alloxan-induced hyperglycemic mice were used in this study. The insulin secretion and glycogen synthesis of the mice were analyzed. At the same time, the gluconeogenesis of the normal mice was also determined. The alloxan-damaged pancreatic beta-cells of the mice were also studied in this paper. After the mice were administered (i.g.) with CCRV, the level of insulin secretion and glycogen synthesis of alloxan-induced hyperglycemic mice elevated (p<0.05, p<0.01) and the gluconeogenesis of the normal mice was inhibited (p<0.01). Also, the alloxan-damaged pancreatic beta-cells of the mice were partly recovered gradually after the mice were administered (i.g.) with CCRV 15 days later. These may account for the causes of CCRV-induced significant decreases of the blood glucose in hyperglycemic mice.

  5. Aportación al conocimiento del género Coprinus Pers. en Catalunya, III. Notas sobre Coprinus tuberosus Quél.

    OpenAIRE

    Vila i Garcia, Jordi; Llistosella i Vidal, Jaume

    1999-01-01

    Se describen y comentan tres muestras de Coprinus tuberosus Qué!., prestando especial atención a la presencia de perisporio en la espora. Se aportan micrograffas al microscopio electrónico de barrido.

  6. Plane Angles: why must the radian have a measure equal to the unity?

    OpenAIRE

    Rocco, Héctor Oscar Di; UNCPBA – Argentina

    2009-01-01

    In this article is discussed a subject that generates controversies inthe teaching of Physics: why the radian must be the correct unitfor the measurements of plane angles. The answer is, in ouropinion, in the concepts of Calculus, where the analytical notionsare independent of the geometrical ones. http://dx.doi.org/10.5007/2175-7941.2009v26n1p136En este artículo se trata de un asunto que genera polémicas en la enseñanza de la Física: por qué el radián debe tener asignada una medida igual ...

  7. Extraction and analysis of the width, gray scale and radian in Chinese signature handwriting.

    Science.gov (United States)

    Chen, Xiaohong

    2015-10-01

    Forensic handwriting examination is a relevant identification process in forensic science. This research obtained ideas from the process of features detection and analysis in forensic handwriting examination. A Chinese signature database was developed and comprised original signatures, freehand imitation forgeries, random forgeries and tracing imitation forgeries. The features of width, gray scale and radian combined with stroke orders were automatically extracted after image processing. A correlation coefficient was used to precisely characterize and express the similarities between signatures. To validate the differences between writers, a multivariate analysis of the variance was employed. The canonical discriminant analysis was performed between the original and non-original signatures; the cross-validation estimated the discriminating power of the width, gray scale and radian data. It is suggested that the extraction and analysis of these properties in Chinese signatures is reasonable. Meanwhile, forensic handwriting examination using the quantitative feature extraction and statistical analysis methods in this research could be performed with a satisfactory result in the discriminant analysis. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

  8. Optimization of the functional expression of Coprinus cinereus peroxidase in Pichia pastoris by varying the host and promoter.

    Science.gov (United States)

    Kim, Su-Jin; Lee, Jeong-Ah; Kim, Yong-Hwan; Song, Bong-Keun

    2009-09-01

    Peroxidase from Coprinus cinereus (CiP) has attracted attention for its high specific activity and broad substrate spectrum compared with other peroxidases. In this study, the functional expression of this peroxidase was successfully achieved in the methylotrophic yeast Pichia pastoris. The expression level of CiP was increased by varying the microbial hosts and the expression promoters. Since a signal sequence, such as the alpha mating factor of Saccharomyces cerevisiae, was placed preceding the cDNA of the CiP coding gene, expressed recombinant CiP (rCiP) was secreted into the culture broth. The Mut+ Pichia pastoris host showed a 3-fold higher peroxidase activity, as well as 2-fold higher growth rate, compared with the Muts Pichia pastoris host. Furthermore, the AOX1 promoter facilitated a 5-fold higher expression of rCiP than did the GAP promoter.

  9. Purification, characterization and evaluation of extracellular peroxidase from two Coprinus species for aqueous phenol treatment.

    Science.gov (United States)

    Ikehata, Keisuke; Buchanan, Ian D; Pickard, Michael A; Smith, Daniel W

    2005-11-01

    Non-ligninolytic fungal peroxidases produced by Coprinus cinereus UAMH 4103 and Coprinus sp. UAMH 10067 were purified, characterized and evaluated as cost-effective alternatives to horseradish peroxidase for aqueous phenol treatment. Purified Coprinus peroxidases exhibited a molecular weight of 36 kDa on matrix-assisted laser desorption ionization time-of-flight mass spectrometry. Although the catalytic properties of the two Coprinus peroxidases were nearly identical in both crude and purified forms, the stabilities were substantially different. The peroxidase from Coprinus sp. UAMH 10067 was more stable at 50 degrees C and under basic conditions (up to pH 10) than the enzyme from C. cinereus UAMH 4103. The former enzyme also performed better at pH 9 than the latter one in aqueous phenol treatment. The phenol removal efficiency of the Coprinus peroxidase was comparable to those of previously studied plant peroxidases. The broader working pH and higher thermal and alkaline stability of the peroxidase from Coprinus sp. UAMH 10067 may be advantageous for its application to industrial wastewater treatment.

  10. Inversion of 5S ribosomal RNA genes within the genus Coprinus.

    Science.gov (United States)

    Cassidy, J R; Pukkila, P J

    1987-01-01

    In the basidiomycete fungus Coprinus cinereus, the 5S ribosomal RNA (rRNA) genes are found in the rDNA repeat which encodes the other three rRNAs (18S, 5.8S, and 26S), and all genes are transcribed in the same direction (Cassidy et al. 1984). To facilitate examination of the rRNA gene organization in additional species, we developed a method to permit determination of the direction of transcription of the rRNA genes using crude preparations of genomic DNA. We used this method to determine the organization of the rDNA in five other members of the order Agaricales (Flammulina velutipes, Agaricus bisporus, Coprinus micaceus, Coprinus atramentarius, and Coprinus comatus). In four, the organization is the same as it is in Coprinus cinereus. However, in Coprinus comatus, the 5S RNA gene is present in the rDNA repeat, but is transcribed in the opposite direction. This alteration in rDNA organization within the genus Coprinus indicates that inverted 5S genes can arise and become propagated through the tandem array of genes. The presence of orientation-dependent promoter elements within the spacer DNA (Elion and Warner 1984) would be expected to place constraints on such inversions.

  11. Optimal Conditions for the Mycelial Growth of Coprinus comatus Strains

    Science.gov (United States)

    Lee, Yun-Hae; Liu, Jun-Jie; Ju, Young-Cheol

    2009-01-01

    The principal objective of this study was to acquire basic data regarding the mycelial growth characteristics for the artificial cultivation of Coprinus comatus. 12 URP primers were employed to evaluate the genetic relationships of C. comatus, and the results were divided into three groups. Among six kinds of mushroom media, MYP medium was selected as the most favorable culture medium for C. comatus. The optimal temperature and pH ranges for the mycelial growth of C. comatus were 23~26℃ and pH 6~8, respectively. The carbon and nitrogen sources for optimal mycelial growth were sucrose and tryptone, respectively. PMID:23983517

  12. ’n Bydrae tot die basidiokarpmorfologie van Coprinus comatus

    Directory of Open Access Journals (Sweden)

    A. Winterboer

    1983-03-01

    Full Text Available Die vorm van die pileus van Coprinus comatus (Müll, ex Fr. S.F. Gray verander tydens ontwikkeling van ovaalvormig na klokvormig. Die hifes besit gespeverbindings en daar kom doliporieë in die dwarswande voor. Daar is twee soorte hifes: breë, opgeblase en dunner draadagtige hifes. Die lamelle bestaan uit drie lae: trama, subhimenium en himenium. Die himeniumlaag is diggepak met basidiums wat twee generasies voorstel, en sistidiums wat tussen die basidiums voorkom. Daar kom vesikels en glikogeenkorrels in die basidium voor. Die spoor is ellipsvormig en het ’n dik wand wat uit verskeie lae opgebou is.

  13. Optimal Conditions for the Mycelial Growth of Coprinus comatus Strains

    OpenAIRE

    Jang, Myoung-Jun; Lee, Yun-Hae; Liu, Jun-Jie; Ju, Young-Cheol

    2009-01-01

    The principal objective of this study was to acquire basic data regarding the mycelial growth characteristics for the artificial cultivation of Coprinus comatus. 12 URP primers were employed to evaluate the genetic relationships of C. comatus, and the results were divided into three groups. Among six kinds of mushroom media, MYP medium was selected as the most favorable culture medium for C. comatus. The optimal temperature and pH ranges for the mycelial growth of C. comatus were 23~26? and p...

  14. Studies on mushroom flavours 2. Flavour compounds in coprinus comatus.

    Science.gov (United States)

    Dijkstra, F Y; Wikén, T O

    1976-01-01

    In an aqueous extract of fruit bodies of Coprinus comatus 3-octanone, 3-octanol, 1-octen-3-ol, 1-octanol, 2-methyl-2-penten-4-olide, 1-dodecanol and caprylic acid were identified conclusively and n-butyric and isobutyric acids preliminarily. Amino-acids, nucleotides and sugars were also determined. A mixture of 37 compounds found in the extract had a stronger flavour than the natural extract. 3-Octanol, 1-octen-3-ol, 1-octanol and 2-methyl-2-penten-4-olide were the volatiles with the strongest flavour. Mass and IR spectra of 2-methyl-2-penten-4-olide are presented.

  15. Optimal Conditions for the Mycelial Growth of Coprinus comatus Strains.

    Science.gov (United States)

    Jang, Myoung-Jun; Lee, Yun-Hae; Liu, Jun-Jie; Ju, Young-Cheol

    2009-06-01

    The principal objective of this study was to acquire basic data regarding the mycelial growth characteristics for the artificial cultivation of Coprinus comatus. 12 URP primers were employed to evaluate the genetic relationships of C. comatus, and the results were divided into three groups. Among six kinds of mushroom media, MYP medium was selected as the most favorable culture medium for C. comatus. The optimal temperature and pH ranges for the mycelial growth of C. comatus were 23~26℃ and pH 6~8, respectively. The carbon and nitrogen sources for optimal mycelial growth were sucrose and tryptone, respectively.

  16. Antioxidant Properties of Cap and Stipe from Coprinus comatus

    OpenAIRE

    Haijuan Nan; Xiaomin Suo; Fei Lu; Bo Li; Bin Li

    2010-01-01

    Coprinus comatus, also called chicken drumstick mushroom, is currently commercially available in China. Hot water and ethanolic extracts were prepared from cap and stipe of C. comatus fruit bodies and their antioxidant properties were studied. Ethanolic extract from stipe showed high antioxidant activity (80.6%) at 1 mg/mL. Reducing power of hot water extracts from cap was 1.653 at 10 mg/mL. Extracts from cap showed better scavenging ability on DPPH (57.9% at 1 mg/mL) than stipe ones. Ethanol...

  17. Pharmacodynamic action of a commercial preparation of the mushroom Coprinus comatus in rats.

    Science.gov (United States)

    Sabo, A; Stilinovic, N; Vukmirovic, S; Bukumiric, Z; Capo, I; Jakovljevic, V

    2010-10-01

    The pharmacodynamic effect of a 7-day oral treatment with a suspension of Coprinus comatus at doses of 0.835 and 1.670 g/kg in rats was studied. Changes in body weight, bile secretion and hypoglycaemic action were examined together with antipyretic activity and paw oedema tests. Such treatments resulted in a significantly lower increase in the body weight of tested animals (15.73 ± 8.36 g/rat in the untreated group, 8.44 ± 8.23 g/rat (p < 0.05) and 3.18 ± 7.93 g/rat (p < 0.05), for C. comatus 0.835 and 1.67 g/kg, respectively). Hypoglycaemic action was evident only in the glucose load test (6.79 ± 0.61 to 9.70 ± 1.16 (p < 0.05) in the untreated group and 6.47 ± 0.35 to 7.27 ± 0.76 for C. comatus 1.67 g/kg). Histological examination of pancreas cross-sections suggested certain protective functions of the mushroom suspension in alloxan poisoning. In the antipyretic test, a significantly lower increase in body temperature was observed in the mushroom-pretreated rats. In the paw oedema test, no decrease in oedema induced by formalin injection was observed following treatment with C. comatus. Copyright © 2010 John Wiley & Sons, Ltd.

  18. Diversity of sesquiterpene synthases in the basidiomycete Coprinus cinereus.

    Science.gov (United States)

    Agger, Sean; Lopez-Gallego, Fernando; Schmidt-Dannert, Claudia

    2009-06-01

    Fungi are a rich source of bioactive secondary metabolites, and mushroom-forming fungi (Agaricomycetes) are especially known for the synthesis of numerous bioactive and often cytotoxic sesquiterpenoid secondary metabolites. Compared with the large number of sesquiterpene synthases identified in plants, less than a handful of unique sesquiterpene synthases have been described from fungi. Here we describe the functional characterization of six sesquiterpene synthases (Cop1 to Cop6) and two terpene-oxidizing cytochrome P450 monooxygenases (Cox1 and Cox2) from Coprinus cinereus. The genes were cloned and, except for cop5, functionally expressed in Escherichia coli and/or Saccharomyces cerevisiae. Cop1 and Cop2 each synthesize germacrene A as the major product. Cop3 was identified as an alpha-muurolene synthase, an enzyme that has not been described previously, while Cop4 synthesizes delta-cadinene as its major product. Cop6 was originally annotated as a trichodiene synthase homologue but instead was found to catalyse the highly specific synthesis of alpha-cuprenene. Coexpression of cop6 and the two monooxygenase genes next to it yields oxygenated alpha-cuprenene derivatives, including cuparophenol, suggesting that these genes encode the enzymes for the biosynthesis of antimicrobial quinone sesquiterpenoids (known as lagopodins) that were previously isolated from C. cinereus and other Coprinus species.

  19. Relationship between deficiency of phosphoglucose isomerase in Coprinus macrorhizus and fruiting body formation.

    Science.gov (United States)

    Nyunoya, H; Ishikawa, T

    1980-01-01

    A mutant (pgi) of Coprinus macrorhizus deficient in phosphoglucose isomerase did not grow on fructose and grew poorly on glucose. The pgi mutation inhibited the formation of monokaryotic and dikaryotic fruiting bodies. PMID:7191420

  20. A new Coprinus from Papua New Guinea sporulating in pure culture

    NARCIS (Netherlands)

    Uljé, C.B.; Aptroot, A.; Iperen, van A.

    1998-01-01

    From Papua New Guinea a new Coprinus is described, which forms basidiocarps and basidiospores in pure culture. It was isolated from material collected on two collecting trips to the area, in 1992 and 1995.

  1. Transformation of multi-component ginkgolide into ginkgolide B by Coprinus comatus

    National Research Council Canada - National Science Library

    Ding, HongXue; Zhang, ZhiCai; Cao, ShengNan; Xu, Yin; Yu, JianGuo

    2015-01-01

    .... Therefore, it is urgent to find a new method to prepare GB. In the current study, we studied the ability and mechanism to transform multi-component ginkgolide into GB by Coprinus comatus in order to enhance the GB yield...

  2. Amino acid pool composition of the basidiomycete Coprinus cinereus.

    Science.gov (United States)

    Ulrich, Cynthia E; Gathman, Allen C; Lilly, Walt W

    2007-11-01

    The leaf-litter fungus Coprinus cinereus maintains a pool of free amino acid in its mycelium. When the organism is grown under conditions of high nitrogen availability with 13.2 mmol.L-1 L-asparagine as the nitrogen source, the primary constituents of this pool are glutamine, alanine, and glutamic acid. Together these 3 amino acids comprise approximately 70% of the pool. Nitrogen deprivation reduces the size of the free amino acid pool by 75%, and neither a high concentration of ammonium nor a protein nitrogen source support a similar pool size as L-asparagine. Nitrogen deprivation also reduces the concentration of glutamine to the pool while increasing glutamate. Concomitant with this shift is a marked increase in mycelial ammonium.

  3. Guanacastane-type diterpenoids with cytotoxic activity from Coprinus plicatilis.

    Science.gov (United States)

    Liu, Yuanzhen; Li, Yaoyao; Ou, Yixin; Xiao, Shuyan; Lu, Chunhua; Zheng, Zhonghui; Shen, Yuemao

    2012-08-01

    Four new guanacastane-type diterpenoids (1-4), together with the known compound, guanacastepene E (5), were isolated from a basidiomycete of the macro-fungi, Coprinus plicatilis 82. Their structures were elucidated on the basis of extensive spectroscopic analyses, including FT-ICR-MS, UV, IR and 1D and 2D NMR experiments. The in vitro cytotoxic activities of all compounds against the human cancer cell lines HepG2, HeLa, MDA-MB-231, BGC-823, HCT 116, and U2OS were evaluated, only compound 1 exhibited significant cytotoxicities with IC(50) values ranging from 1.2 to 6.0 μM. Copyright © 2012 Elsevier Ltd. All rights reserved.

  4. Natural units in physics, and the curious case of the radian

    Science.gov (United States)

    Quincey, Paul

    2016-11-01

    Sets of natural units, like ‘atomic units’, are sometimes used to simplify the equations of physics. This choice of units can be seen as a way of showing the relationships between quantities in their simplest form, in specialised situations, while still being correct. The unit system used for teaching, the SI, is instead designed primarily to be a set of units that can be applied consistently across all areas of science, commerce and industry. The SI also gives priority to emphasising the distinction between different quantities, by (most of the time) giving different units to different quantities, rather than producing the simplest possible equations. It is not widely appreciated that the SI treats the radian as the natural unit for angle, the only unit to be treated this way, with consequent issues for clear distinctions between quantities involving angles. The system that the SI would become if this anomaly were removed is presented. Rather than advocating this major change to the SI itself, it is proposed to highlight the existence of the underlying system to clarify how angles are handled physics, to make it easier to include angles in software calculations, and as an example of how the choice of unit system affects the equations we use.

  5. Magnetic resonance spectral characterization of the heme active site of Coprinus cinereus peroxidase

    Energy Technology Data Exchange (ETDEWEB)

    Lukat, G.S.; Rodgers, K.R.; Jabro, M.N.; Goff, H.M. (Univ. of Iowa, Iowa City (USA))

    1989-04-18

    Examination of the peroxidase isolated from the inkcap Basidiomycete Coprinus cinereus shows that the 42,000-dalton enzyme contains a protoheme IX prosthetic group. Reactivity assays and the electronic absorption spectra of native Coprinus peroxidase and several of its ligand complexes indicate that this enzyme has characteristics similar to those reported for horseradish peroxidase. In this paper, the authors characterize the H{sub 2}O{sub 2}-oxidized forms of Coprinus peroxidase compounds I, II, and III by electronic absorption and magnetic resonance spectroscopies. Electron paramagnetic resonance (EPR) and nuclear magnetic resonance (NMR) studies of this Coprinus peroxidase indicate the presence of high-spin Fe(III) in the native protein and a number of differences between the heme site of Coprinus peroxidase and horseradish peroxidase. Carbon-13 (of the ferrous CO adduct) and nitrogen-15 (of the cyanide complex) NMR studies together with proton NMR studies of the native and cyanide-complexed Caprinus peroxidase are consistent with coordination of a proximal histidine ligand. The EPR spectrum of the ferrous NO complex is also reported. Protein reconstitution with deuterated hemin has facilitated the assignment of the heme methyl resonances in the proton NMR spectrum.

  6. Antioxidant properties of cap and stipe from Coprinus comatus.

    Science.gov (United States)

    Li, Bo; Lu, Fei; Suo, Xiaomin; Nan, Haijuan; Li, Bin

    2010-03-09

    Coprinus comatus, also called chicken drumstick mushroom, is currently commercially available in China. Hot water and ethanolic extracts were prepared from cap and stipe of C. comatus fruit bodies and their antioxidant properties were studied. Ethanolic extract from stipe showed high antioxidant activity (80.6%) at 1 mg/mL. Reducing power of hot water extracts from cap was 1.653 at 10 mg/mL. Extracts from cap showed better scavenging ability on DPPH (57.9% at 1 mg/mL) than stipe ones. Ethanolic extracts were more effective in scavenging ability on hydroxyl radicals (57.4-61.3% at 5 mg/mL) than hot water extracts. Ethanolic extracts showed moderate scavenging ability on superoxide radicals (46.3-47.0% at 20 mg/mL). Naturally occurring antioxidant components including total phenols (3.60-20.00 mg/g), tocopherols (0.58-11.93 mg/g), flavonoids (0.19-3.52 mg/g) and polysaccharides (58.52-547.86 mg/g) were found in the extracts. Overall, extracts from cap were more effective for the antioxidant properties assayed.

  7. [Separation, purification and antioxidant activity of polysaccharide from Coprinus comatus].

    Science.gov (United States)

    Xu, Nü; Zhang, Tianzhen; Chen, Xufeng; Zhang, Hao; Wang, Rufu

    2017-05-25

    We compared the ways of deproteinization for crude polysaccharides of Coprinus comatus, and finally selected Sevage method as the optimal method. Two main fractions of Ccp-I-A and Ccp-I-B were obtained after DEAE-52 cellulose and Sephadex G-200 chromatography, both were white-floc, soluble in water, insoluble in absolute ethyl alcohol, acetone and other organic solvents. Additionally, Fehling reagent, CTAB, Sulphuric acid-carbazole, I-KI and FeCl₃ reaction were all negative. GC analysis showed Ccp-I-A was composed of mannitose, glucose and galactose in molar ratios of 2.03:9.52:1, whereas Ccp-I-B was composed of fucose and galactose with molar ratios of 1:5.21. Antioxidant activity test showed that Ccp-I-A and Ccp-I-B had good scavenging abilities on DPPH and ·OH. Compared to Ccp-I-B,the scavenging activity of Ccp-I-A was much stronger, and the scavenging rate could reach 72.1% and 55.3% respectively when the concentration was 300 μg/mL.

  8. The origin of multiple B mating specificities in Coprinus cinereus.

    Science.gov (United States)

    Riquelme, Meritxell; Challen, Michael P; Casselton, Lorna A; Brown, Andrew J

    2005-07-01

    Mushrooms, such as Coprinus cinereus, possess large families of pheromones and G-protein-coupled receptors that are sequestered at the B mating-type locus and whose function is to confer vast numbers of different mating types. This ability results from complex patterns of cognate and noncognate pheromone/receptor pairings, which potentially offer a unique insight into the molecular interaction between receptor and ligand. In this study we have identified many more members of these families by molecular analysis of strains collected worldwide. There are three groups of genes at each B locus. We have identified two alleles of group 1, five alleles of group 2, and seven alleles of group 3, encoding in total 14 different receptors and 29 different pheromones. The specificity of many newly identified alleles was determined by transformation analysis. One striking finding was that receptors fall into groups based on sequence homology but these do not correspond to the groups defined by position, indicating that complex evolutionary processes gave rise to the B loci. While additional allelic versions may occur in nature, the number of B specificities possible by combination of the alleles that we describe is 70, close to previous estimates based on population analysis.

  9. The laccase gene family in Coprinopsis cinerea (Coprinus cinereus).

    Science.gov (United States)

    Hoegger, Patrik J; Navarro-González, Monica; Kilaru, Sreedhar; Hoffmann, Matthias; Westbrook, Elisha D; Kües, Ursula

    2004-02-01

    In this study, we isolated and sequenced eight non-allelic laccase genes from Coprinopsis cinerea ( Coprinus cinereus) homokaryon AmutBmut. These eight genes represent the largest laccase gene family identified so far in a single haploid fungal genome. We analyzed the phylogenetic relationships between these genes by intron positions, amino acid sequence conservation and similarities in promoter sequences. All deduced protein products have the laccase signature sequences L1-L4, the typical conserved cysteine and the ten histidine residues which are ligands in the two laccase copper-binding centers, T1 and T2/T3. Proteins Lcc2 and Lcc3 of Coprinopsis cinerea are most similar to the acidic, membrane-associated laccase CLAC2 from Coprinellus congregatus implicated in neutralization of acidic medium. All other laccases from the saprophyte Coprinopsis cinerea, including the well described enzyme Lcc1, form a cluster separate from these three enzymes and from various laccases of wood-rotting and plant-pathogenic basidiomycetes.

  10. Inheritance of Chromosome-Length Polymorphisms in Coprinus Cinereus

    Science.gov (United States)

    Zolan, M. E.; Heyler, N. K.; Stassen, N. Y.

    1994-01-01

    We have investigated the inheritance of chromosome-length polymorphisms in the basidiomycete Coprinus cinereus. The electrophoretic karyotypes of interfertile strains of C. cinereus are strikingly different, and crosses between strains with different karyotypes yield progeny with chromosomes of new sizes. Repeated backcrossing of a mutant to one parent often stabilizes the mutant chromosome at a unique size; this then becomes a chromosome-length polymorphism marker for that mutant gene. A comparison of mutant strains, their wild-type progenitor, and backcrossed strains revealed that these marker chromosomes are not caused by the initial mutagenic treatment and are found only in progeny of crosses between strains with polymorphic chromosomes. Thus, they are most likely formed by meiotic recombination. For the rad12 gene, the marker chromosome can further recombine to become the size of the homolog of the backcross parent. For the rad3 gene, both ectopic and homologous recombination events are likely involved in the generation of the marker chromosomes. As predicted by a recombination model, a cross to a new wild-type parent can change the size of a mutant marker chromosome. Therefore, changes in chromosome length are a common and prominent feature of the genome of this sexual fungus, and a variety of karyotypes is tolerated by the organism. PMID:7914506

  11. De Novo Methylation of Repeated Sequences in Coprinus Cinereus

    Science.gov (United States)

    Freedman, T.; Pukkila, P. J.

    1993-01-01

    We have examined the stability of duplicated DNA sequences in the sexual phase of the life cycle of the basidiomycete fungus, Coprinus cinereus. We observed premeiotic de novo methylation in haploid nuclei containing either a triplication, a tandem duplication, or an ectopic duplication. Methylation changes were not observed in unique sequences. Repeated sequences underwent methylation changes during the dikaryotic stage. In one cross, 27% of the segregants exhibited methylation-directed gene inactivation. However, all auxotrophs eventually reverted to prototrophy. C to T transition mutations were not observed in this study. Our studies also revealed one inversion that occurred in 50% of the segregants in a single triplication cross, and a single pop-out event that occurred during vegetative growth. These alterations were similar to changes reported in experiments with duplicated sequences in Neurospora crassa and Ascobolus immersus. However, significant differences were also noted. First, the extent of methylation was much less in C. cinereus than in the other two fungi. Second, CpG sequences appeared to be the preferred targets of methylation. PMID:8244000

  12. Inhibition of the androgen receptor activity by Coprinus comatus substances.

    Science.gov (United States)

    Dotan, Nesly; Wasser, Solomon P; Mahajna, Jamal

    2011-11-01

    Prostatic adenocarcinoma is the second leading cause of death from cancer in Western men. The common prostate cancer treatments are effective in the early stages; however, advanced prostate cancer is resilient to most of these treatments. Altered androgen receptor (AR) activity caused by point mutations or signaling mechanisms that regulate AR function has been proposed as a key mechanism in the transition to the androgen-independent stage. Our previous results demonstrated that hexane extract prepared from Coprinus comatus (C. comatus) strain 734 was able to interfere with AR activity. The current study was made to further evaluate the antiandrogenic activity of the C. comatus mushroom strain 734. Activity-guided chromatography was conducted and 2 active fractions, F-32-and F-33, were found to contain substances that were able to inhibit AR-mediated reporter activity and reduce the levels of AR and prostate-specific antigen (PSA) transcripts in LNCaP cells. Fraction F-32 also inhibited the proliferation and clonigenicity of LNCaP cells. Furthermore, F-32 was able to inhibit the binding of AR to the PSA enhancer region and to inhibit Akt-mediated AR phosphorylation at Ser 213. This study illustrated the potential of substances from the C. comatus mushroom to serve as natural antiandrogenic modulators for the treatment of prostatic disorders.

  13. Antioxidant Properties of Cap and Stipe from Coprinus comatus

    Directory of Open Access Journals (Sweden)

    Haijuan Nan

    2010-03-01

    Full Text Available Coprinus comatus, also called chicken drumstick mushroom, is currently commercially available in China. Hot water and ethanolic extracts were prepared from cap and stipe of C. comatus fruit bodies and their antioxidant properties were studied. Ethanolic extract from stipe showed high antioxidant activity (80.6% at 1 mg/mL. Reducing power of hot water extracts from cap was 1.653 at 10 mg/mL. Extracts from cap showed better scavenging ability on DPPH (57.9% at 1 mg/mL than stipe ones. Ethanolic extracts were more effective in scavenging ability on hydroxyl radicals (57.4–61.3% at 5 mg/mL than hot water extracts. Ethanolic extracts showed moderate scavenging ability on superoxide radicals (46.3–47.0% at 20 mg/mL. Naturally occurring antioxidant components including total phenols (3.60–20.00 mg/g, tocopherols (0.58–11.93 mg/g, flavonoids (0.19–3.52 mg/g and polysaccharides (58.52–547.86 mg/g were found in the extracts. Overall, extracts from cap were more effective for the antioxidant properties assayed.

  14. Coprinus comatus and Ganoderma lucidum interfere with androgen receptor function in LNCaP prostate cancer cells.

    Science.gov (United States)

    Zaidman, Ben-Zion; Wasser, Solomon P; Nevo, Eviatar; Mahajna, Jamal

    2008-06-01

    In this study, we screened a total of 201 diethyl ether, ethanol, and ethyl acetate fungal Basidiomycetes extracts for anti-androgenic activity. Based on our screened results in combination with the selective inhibition of prostate cancer LNCaP cells, we selected Coprinus comatus and Ganoderma lucidum for further evaluation. We demonstrated that ethanol and ethyl acetate extracts from C. comatus and G. lucidum, respectively, selectively inhibit dihydrotestosterone-induced LNCaP cell viability, suppress levels of secreted prostate-specific antigen in a dose-dependent manner, and cause a G1 phase arrest in LNCaP, but not in DU 145 and PC-3 cells. For the first time, to the best of our knowledge, we demonstrated that C. comatus and G. lucidum decreased androgen and glucocorticoide receptors transcriptional activity in breast cancer MDA-kb2 cells in a dose-dependent manner, and suppressed androgen receptor (AR) protein level in LNCaP and MDA-kb2 cells. Our findings suggest that AR and non-AR mediated mechanisms underlie the effects of C. comatus and G. lucidum.

  15. [Enhancement of Coprinus cinereus peroxidase in Pichia pastoris by co-expression chaperone PDI and Ero1].

    Science.gov (United States)

    Chen, Fei; Hu, Meirong; Jiang, Xianzhang; Tao, Yong; Huang, Jianzhong

    2015-12-01

    The 1,095 bp gene encoding peroxidase from Coprinus cinereus was synthesized and integrated into the genome of Pichia pastoris with a highly inducible alcohol oxidase. The recombinant CiP (rCiP) fused with the a-mating factor per-pro leader sequence derived from Saccharomyces cerevisiae was secreted into the culture medium and identified as the target protein by mass spectrometry, confirming that a C. cinereus peroxidase (CiP) was successfully expressed in P. pastoris. The endoplasmic reticulum oxidoreductase 1 (Ero1) and protein disulfide isomerase (PDI) were co-expressed with rCiP separately and simultaneously. Compared with the wild type, overexpression of PDI and Erol-PDI increaseed Cip activity in 2.43 and 2.6 fold and their activity reached 316 U/mL and 340 U/mL respectively. The strains co-expressed with Erol-PDI was used to high density fermentation, and their activity reached 3,379 U/mL, which was higher than previously reported of 1,200 U/mL.

  16. Structural investigation of water-soluble polysaccharides extracted from the fruit bodies of Coprinus comatus

    NARCIS (Netherlands)

    Li, Bo; Dobruchowska, Justyna M.; Gerwig, Gerrit J.; Dijkhuizen, Lubbert; Kamerling, Johannis P.

    2013-01-01

    Water-soluble polysaccharide material, extracted from the stipes of the fruit bodies of Coprinus comatus by hot water, was fractionated by sequential weak anion-exchange and size-exclusion chromatography. The relevant fractions were subjected to structural analysis, including (D/L)

  17. Consumption of Coprinus comatus polysaccharide extract causes recovery of alcoholic liver damage in rats

    NARCIS (Netherlands)

    Ozalp, F.O.; Canbek, M.; Yamac, M.; Kanbak, G.; Griensven, van L.J.L.D.; Uyanoglu, M.; Senturk, H.; Karlkava, K.; Oglakci, A.

    2014-01-01

    Excess use of alcohol is known to be associated with liver diseases such as fatty liver, alcoholic hepatitis, and cirrhosis. Various practices may be applied to prevent or treat the damage caused by chronic alcoholism. Coprinus comatus (O.F. Müll.) Pers. (Agaricaceae) is a macrofungus that has been

  18. Life History and Developmental Processes in the Basidiomycete Coprinus cinereus

    Science.gov (United States)

    Kües, Ursula

    2000-01-01

    Coprinus cinereus has two main types of mycelia, the asexual monokaryon and the sexual dikaryon, formed by fusion of compatible monokaryons. Syngamy (plasmogamy) and karyogamy are spatially and temporally separated, which is typical for basidiomycetous fungi. This property of the dikaryon enables an easy exchange of nuclear partners in further dikaryotic-monokaryotic and dikaryotic-dikaryotic mycelial fusions. Fruiting bodies normally develop on the dikaryon, and the cytological process of fruiting-body development has been described in its principles. Within the specialized basidia, present within the gills of the fruiting bodies, karyogamy occurs in a synchronized manner. It is directly followed by meiosis and by the production of the meiotic basidiospores. The synchrony of karyogamy and meiosis has made the fungus a classical object to study meiotic cytology and recombination. Several genes involved in these processes have been identified. Both monokaryons and dikaryons can form multicellular resting bodies (sclerotia) and different types of mitotic spores, the small uninucleate aerial oidia, and, within submerged mycelium, the large thick-walled chlamydospores. The decision about whether a structure will be formed is made on the basis of environmental signals (light, temperature, humidity, and nutrients). Of the intrinsic factors that control development, the products of the two mating type loci are most important. Mutant complementation and PCR approaches identified further genes which possibly link the two mating-type pathways with each other and with nutritional regulation, for example with the cAMP signaling pathway. Among genes specifically expressed within the fruiting body are those for two galectins, β-galactoside binding lectins that probably act in hyphal aggregation. These genes serve as molecular markers to study development in wild-type and mutant strains. The isolation of genes for potential non-DNA methyltransferases, needed for tissue formation

  19. Nutrients and non-nutrients composition and bioactivity of wild and cultivated Coprinus comatus (O.F.Müll.) Pers.

    NARCIS (Netherlands)

    Stojkovic, D.; Reis, F.S.; Barros, L.; Glamoclija, J.; Ciric, A.; Griensven, van L.J.L.D.; Sokovic, M.; Ferreira, I.C.F.R.

    2013-01-01

    Mushrooms have been reported as sources of biomolecules with various potential. Coprinus comatus was studied to obtain information about this species, comparing cultivated and wild samples. Free sugars, fatty acids, tocopherols, organic acids and phenolic acids were analyzed by chromatographic

  20. Vanadium uptake by biomass of Coprinus comatus and their effect on hyperglycemic mice.

    Science.gov (United States)

    Han, Chunchao; Cui, Bo; Wang, Yingzi

    2008-07-01

    The Vanadium uptake by Coprinus comatus and their co-effect on hyperglycemic mice were studied. By fermentation and AAS analysis, the optimal concentration of vanadium in medium was 0.4%, and the content of vanadium accumulated in the mycelia was 3,528.0 microg/g. At the concentration of 0.4%, the vanadium-associated toxicity was reduced, and its anti-diabetic effects were maintained.

  1. Anti-Oxidative Activity of An Aqueous Suspension of Commercial Preparation of The Mushroom Coprinus comatus

    OpenAIRE

    Mira Popović; Saša Vukmirović; Nebojša Stilinović; Ivan Čapo; Vida Jakovljević

    2010-01-01

    In this study the effects of an aqueous suspension of a commercial preparation of the mushroom Coprinus comatus on oxidative stress induced in rats by alloxane and carbon tetrachloride was examined. The effects were estimated from changes in the biochemical parameters (xanthine oxidase, glutathione peroxidase and catalase activity, reduced glutathione content, and extent of lipid peroxidation) of liver homogenate as well as histological changes in the liver of the rats treated with alloxane a...

  2. Viability of mycelial growth of Coprinus Comatus in culture medium based on organic residues

    OpenAIRE

    de Almeida, Lais Benes; Sales-Campos, Ceci; Melo de Carvalho, Cristiane Suely; de Almeida Minhoni, Marli Teixeira [UNESP; Nogueira de Andrade, Meire Cristina

    2012-01-01

    The objective of this work was to evaluate the mycelial growth of the Coprinus comatus strain CCO 01/01 in culture based on organic residues of Saccharum officinarum (sugarcane bagasse), Citrus sinensis (orange bagasse), Ananas comosus (pineapple residues) and Musa sp. (banana leaf), supplemented with wheat bran in the proportions of 0, 10 and 20%, kept at 27 degrees C. The mycelial growth of C. comatus was evaluated daily by measurement of the diameter of the colony during seven days of incu...

  3. Determination of biodiversity of Coprinus comatus using genotyping and metabolic profiling tools.

    Science.gov (United States)

    Pawlik, Anna; Malinowska, Anna; Siwulski, Marek; Frąc, Magdalena; Rogalski, Jerzy; Janusz, Grzegorz

    2015-01-01

    Coprinus comatus strains (CCMs) originating from Poland were identified using ITS region sequencing. Based on the sequences obtained, the genetic relationship between the CCM strains was determined and a clear separation of all strains into two main clusters was obtained. The Coprinus strains were also genetically characterized for the first time by the AFLP technique. The analysis showed that the CCMs separated into four main clusters and a high complication of a UPGMA-based dendrogram was achieved. C. comatus strains included in the analysis displayed an AFLP profile similarity level in the range from 44 to 66%. The highest similarity coefficient, 0.490, was found between CCM12 and CCM13, and the lowest (0.202) between the CCM2 and CCM5 isolates. Biolog FF MicroPlates were applied to obtain data on utilization of 95 carbon sources and mycelial growth. The analysis allowed comparison of the functional diversity of the CCM strains and revealed a broad variability within the analyzed Coprinus species based on substrate utilization profiles. Significant differences (2-48) have been shown in the substrate richness values. The Biolog experiments proved to be a good profiling technology for studying the diversity in shaggy manes due to metabolic differences and demonstrated that all the strains might be considered individually. It is evident that the strain metabolic grouping does not correlate with the grouping based on the ITS sequences and AFLP profiles, however, some similarities may be observed.

  4. Isolation and biological activity of triglycerides of the fermented mushroom of Coprinus Comatus

    Directory of Open Access Journals (Sweden)

    Ren Jun

    2012-04-01

    Full Text Available Abstract Background Although many physiological functions of Coprinus comatus have been reported, there has been no report on the antinociceptive activity of Coprinus comatus. Therefore, the objective of the present study is to demonstrate the production, isolation, and biological properties of triglycerides (TFC of the fermented mushroom of Coprinus comatus. Methods The effects of TFC on cytokines levels, total antioxidant activity, antinociceptive effects in vivo, LD50 and tactile hyperalgesia were analyzed respectively. Results TFC treatment decreased the levels of cytokines and total antioxidant status (TAOS and inhibited the acetic acid-induced abdominal constrictions in mice. In addition, TFC reduced CFA-induced tactile hyperalgesia in a dose-dependent manner and the LD50 of TFC was determined to be 400 mg/kg. However, TFC did not significantly inhibit the reaction time to thermal stimuli in the hot-plate test. Conclusions TFC showed anti-inflammatory, antioxidant, peripheral antinociceptive and antihyperalgesic activity in various models of inflammatory pain. The data suggest that TFC may be a viable treatment option for inflammatory pain.

  5. Isolation and biological activity of triglycerides of the fermented mushroom of Coprinus Comatus.

    Science.gov (United States)

    Ren, Jun; Shi, Jin-Li; Han, Chun-Chao; Liu, Zhen-Quan; Guo, Jian-You

    2012-04-24

    Although many physiological functions of Coprinus comatus have been reported, there has been no report on the antinociceptive activity of Coprinus comatus. Therefore, the objective of the present study is to demonstrate the production, isolation, and biological properties of triglycerides (TFC) of the fermented mushroom of Coprinus comatus. The effects of TFC on cytokines levels, total antioxidant activity, antinociceptive effects in vivo, LD50 and tactile hyperalgesia were analyzed respectively. TFC treatment decreased the levels of cytokines and total antioxidant status (TAOS) and inhibited the acetic acid-induced abdominal constrictions in mice. In addition, TFC reduced CFA-induced tactile hyperalgesia in a dose-dependent manner and the LD50 of TFC was determined to be 400 mg/kg. However, TFC did not significantly inhibit the reaction time to thermal stimuli in the hot-plate test. TFC showed anti-inflammatory, antioxidant, peripheral antinociceptive and antihyperalgesic activity in various models of inflammatory pain. The data suggest that TFC may be a viable treatment option for inflammatory pain.

  6. Decolorization of the anthraquinone dye Cibacron Blue 3G-A with immobilized Coprinus cinereus in fluidized bed bioreactor.

    Science.gov (United States)

    Moutaouakkil, A; Blaghen, M

    2011-01-01

    Coprinus cinereus, which was able to decolorize the anthraquinone dye Cibacron Blue 3G-A (CB) enzymatically, was used as a biocatalyst for the decolorization of synthetic solutions containing this reactive dye. Coprinus cinereus was immobilized in both calcium alginate and polyacrylamide gels, and was used for the decolorization of CB from synthetic water by using a fluidized bed bioreactor. The highest specific decolorization rate was obtained when Coprinus cinereus was entrapped in calcium alginate beads, and was of about 3.84 mg g(-1) h(-1) with a 50% conversion time (t1/2) of about 2.60 h. Moreover, immobilized fungal biomass in calcium alginate continuously decolorized CB even after 7 repeated experiments without significant loss of activity, while polyacrylamide-immobilized fungal biomass retained only 67% of its original activity. The effects of some physicochemical parameters such as temperature, pH and dye concentration on decolorization performance of isolated fungal strain were also investigated.

  7. The role of calcium accumulation and the cytoskeleton in the perception and response of Coprinus cinereus to gravity

    Science.gov (United States)

    Frazer, L. Novak; Moore, D.

    1996-03-01

    The role of Ca^2+ in the gravitropic perception and/or response mechanism of Coprinus cinereus was examined by treating stipes with inhibitors of Ca^2+ transport and calmodulin. Inhibitors had no effect on gravity perception but significantly diminished gravitropism. It is concluded that, under the conditions tested, Ca^2+ is not involved in gravity perception by Coprinus stipes, but does contribute to transduction of the gravitropic impulse. The results would be consistent with regulation of the gravitropic bending process requiring accumulation of Ca^2+ within a membrane-bound compartment. Treatment of stipes with an actin inhibitor caused a significantly delayed response, a result not observed with the Ca^2+ inhibitors. This suggests that cytoskeletal elements may be involved directly in perception of gravity by Coprinus stipes while Ca^2+-mediated signal transduction may be involved in directing growth differentials.

  8. Resíduos de bananeira como substrato base para o cultivo in vitro de Coprinus comatusBanana tree residues as base substrate for the in vitro cultivation of Coprinus comatus

    OpenAIRE

    Almeida, Laís Bentes de; UFAM; Sales-Campos, Ceci; INPA; Carvalho, Cristiane Suely Melo de; UFAM; Minhoni, Marli Teixeira de Almeida; UNESP; Andrade, Meire Cristina Nogueira de; UNESP

    2013-01-01

    O Coprinus comatus é um fungo lignícola e comestível que tem demonstrado grande potencial para sua exploração comercial, por se desenvolver facilmente nos mais diferentes resíduos, entre eles a folha de bananeira. Assim, avaliou-se o crescimento micelial de Coprinus comatus em meios de cultura à base de folhas das diferentes cultivares de bananeira: Thap-Maeo, Prata-Anã Pelipita e Caipira, com 20% de suplementação com farelo de trigo, soja e arroz. Em câmara de fluxo laminar, discos de 7 mm d...

  9. Thermal performance of 625-kg/cu m elastomeric ablative materials on spherically blunted 0.44-radian cones

    Science.gov (United States)

    Champman, A. J.

    1972-01-01

    Spherically blunted 0.44-radian (25 deg) half-angle conical models coated with elastomeric ablative materials were tested in supersonic arc-heated wind tunnels to evaluate performance of the ablators over a range of conditions typical of lifting entry. Four test conditions were combinations of stagnation point-heat transfer rates of 2.3 and 4.5 MW/m2 and stagnation pressures of 20 and 2kN/m2. Afterbody values of heat transfer rate and pressure were 0.05 to 0.20 of stagnation point values. Stagnation enthalpy varied from 4.4 to 25 MJ/kg (1900 to 11000 Btu/lbm) and free-stream Mach number was in a range from 3.5 to 4. Ablative materials retained the spherical nose shape throughout tests at the lower heat transfer level, but receded, assuming a flattened nose shape, during tests at the high heat transfer level. The residue layer that formed on the conical after-body was weak, friable, and extensively cracked. The reference ablative material, which contained phenolic microspheres, generally retained the conical shape on the model afterbody. However, a modified ablator, in which phenolic microspheres were replaced with silica microspheres, deformed and separated from the undegraded material, and thereby produced a very uneven surface. Substrate temperatures and ablator recession were in good agreement with values computed by a numerical analysis.

  10. Anthopleura radians, a new species of sea anemone (Cnidaria: Actiniaria: Actiniidae from northern Chile, with comments on other species of the genus from the South Pacific Ocean

    Directory of Open Access Journals (Sweden)

    Carlos Spano

    2017-05-01

    Full Text Available A new species of sea anemone, Anthopleura radians n. sp., is described from the intertidal zone of northern Chile and the taxonomic status of the other Anthopleura species from the South Pacific are discussed. A. radians n. sp. is characterized by a yellow-whitish and brown checkerboard-like pattern on the oral disc, adhesive verrucae along the entire column and a series of marginal projections, each bearing a brightly-colored acrorhagus on the oral surface. This is the seventh species of Anthopleura described from the South Pacific Ocean; each one distinguished by a particular combination of differences related to their coloration pattern, presence of zooxanthellae, cnidae, and mode of reproduction. Some of these species have not been reported since their original description and thus require to be taxonomically validated. A. hermaphroditica and A. aureoradiata are synonyms considering the lack of differences seen between live specimens, museum collections and published records. A. radians could also be a junior synonym of A. minima, however, no type material was found for testing this hypothesis. Furthermore, it is crucial to designate neotypes for A. inconspicua, A. rosea and A. minima since there are no name-bearing types reported for these species.

  11. Coprinus comatus Cap Inhibits Adipocyte Differentiation via Regulation of PPARγ and Akt Signaling Pathway

    OpenAIRE

    Park, Hyoung Joon; Yun, Jisoo; Jang, Sun-Hee; Kang, Suk Nam; Jeon, Beong-Sam; Ko, Yeoung-Gyu; Kim, Hong-Duck; Won, Chung-Kil; Kim, Gon-Sup; Cho, Jae-Hyeon

    2014-01-01

    This study assessed the effects of Coprinus comatus cap (CCC) on adipogenesis in 3T3-L1 adipocytes and the effects of CCC on the development of diet-induced obesity in rats. Here, we showed that the CCC has an inhibitory effect on the adipocyte differentiation of 3T3-L1 cells, resulting in a significant decrease in lipid accumulation through the downregulation of several adipocyte specific-transcription factors, including CCAAT/enhancer binding protein β, C/EBPδ, and peroxisome proliferator-a...

  12. Xylaria coprinicola, a new species that antagonizes cultivation of Coprinus comatus in China.

    Science.gov (United States)

    Ju, Yu-Ming; Hsieh, Huei-Mei; He, Xin-Sheng

    2011-01-01

    The species known in China as the chicken-claw fungus is described as a new species, Xylaria coprinicola. This species is known as an antagonist of cultivation of the edible mushroom Coprinus comatus. Stromata of X. coprinicola are cylindrical, terminate in a sterile apex and arise in fascicles from a relative large submerged base; perithecia are immersed and have conspicuously conical ostiolar openings; ascospores are minute. Phylogenetic analyses based on combined partial sequences of rpb2, β-tub and α-act genes showed that X. coprinicola is closely related to those Xylaria species exclusively associated with termite nests.

  13. New to Poland species of the broadly defined genus Coprinus (Basidiomycota, Agaricomycotina

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    Błażej Gierczyk

    2014-12-01

    Full Text Available The paper presents a list of 19 coprinoid fungi, found in Poland, which have not been reported earlier from this area: Coprinellus bisporiger, C. dilectus, C. heterothrix, C. radicellus, Coprinopsis annulopora, C. bellulus, C. candidolanata, C. cinereofloccosa, C. coniophora, C. goudensis, C. idae, C. iocularis, C. krieglsteineri, C. pachyderma, C. phlyctidospora, C. rugosobispora, C. scobicola, C. spilospora, Coprinus palmeranus. Illustrations and short descriptions of the species, based on the specimens examined and literature data, are given.

  14. Patterns of Mating and Mitochondrial DNA Inheritance in the Agaric Basidiomycete Coprinus Cinereus

    Science.gov (United States)

    May, G.; Taylor, J. W.

    1988-01-01

    Patterns of mating and mitochondrial DNA (mtDNA) inheritance were investigated for the Basidiomycete, Coprinus cinereus in order to better understand the relationship of reproductive biology and mtDNA evolution in fungi. Results showed that the unique mating system of basidiomycetes can lead to the formation of mitochondrial mosaics (i.e., colonies composed of sectors differing in mtDNA). Mitochondria do not migrate along with nuclei during mating. Intracellular mixed or recombinant mtDNA molecules were not observed. Interestingly, it was found that mating asymmetry, caused by nonreciprocal nuclear migration, may be an important part of the reproductive biology of C. cinereus. PMID:2834264

  15. Structural elucidation of a neutral fucogalactan from the mycelium of Coprinus comatus.

    Science.gov (United States)

    Fan, JunMin; Zhang, JingSong; Tang, QingJiu; Liu, YanFang; Zhang, AnQiang; Pan, YingJie

    2006-07-03

    A water-soluble fucogalactan (CMP3), with a molecular mass of 1.03 x 10(4) Da as determined by high-performance size-exclusion chromatography (HPSEC), was obtained from the crude intracellular polysaccharide of Coprinus comatus mycelium. Its chemical structure was characterized by sugar and methylation analysis along with 1H and 13C NMR spectroscopy, including NOESY and HMBC experiments for linkage and sequence analysis. The polysaccharide is composed of a pentasaccharide repeating unit with the following structure: [structure:see text].

  16. Some morphological and physiological aspects and cultivation of Coprinus comatus (O. F. Mull.) Gray

    OpenAIRE

    Petcharat, V.; Chaiyama, V; Kritsaneepaiboon, P.

    2007-01-01

    Some morphological and physiological aspects and cultivation of Coprinus comatus (O.F. Mull.) Gray were investigated. Malt extract agar turned out to be the best in supporting the mycelial growth of C.comatus. Mannose and maltose were the best carbon sources in supporting mycelial growth. C. comatus utilized peptone and NH4NO3 better than other nitrogen sources. The optimum temperature and pH on maltextract agar were 25oC and 6, respectively. Light retarded mycelial growth of C. comatus. For ...

  17. Mercury bio-extraction by fungus Coprinus comatus: a possible bioindicator and mycoremediator of polluted soils?

    OpenAIRE

    Falandysz, Jerzy

    2015-01-01

    The Shaggy Ink Cap (Coprinus comatus), which is a common in wild in northern hemisphere was examined in field for potential to be used as possible bio-extractor of Hg from polluted grounds but also as possible bioindicator of urban soils (roadside, barren lands, lawns) pollution with Hg. The contents of Hg in caps and stipes of C. comatus from the grounds examined in this study correlated positively with the levels of soil contamination. Analysis of sets of data available worldwide on Hg in C...

  18. Self-compatible B mutants in coprinus with altered pheromone-receptor specificities.

    Science.gov (United States)

    Olesnicky, N S; Brown, A J; Honda, Y; Dyos, S L; Dowell, S J; Casselton, L A

    2000-01-01

    A successful mating in the mushroom Coprinus cinereus brings together a compatible complement of pheromones and G-protein-coupled receptors encoded by multiallelic genes at the B mating-type locus. Rare B gene mutations lead to constitutive activation of B-regulated development without the need for mating. Here we characterize a mutation that arose in the B6 locus and show that it generates a mutant receptor with a single amino acid substitution (R96H) at the intracellular end of transmembrane domain III. Using a heterologous yeast assay and synthetic pheromones we show that the mutation does not make the receptor constitutively active but permits it to respond inappropriately to a normally incompatible pheromone encoded within the same B6 locus. Parallel experiments carried out in Coprinus showed that a F67W substitution in this same pheromone enabled it to activate the normally incompatible wild-type receptor. Together, our experiments show that a single amino acid replacement in either pheromone or receptor can deregulate the specificity of ligand-receptor recognition and confer a self-compatible B phenotype. In addition, we use the yeast assay to demonstrate that different receptors and pheromones found at a single B locus belong to discrete subfamilies within which receptor activation cannot normally occur. PMID:11063682

  19. Anti-Oxidative Activity of An Aqueous Suspension of Commercial Preparation of The Mushroom Coprinus comatus

    Directory of Open Access Journals (Sweden)

    Mira Popović

    2010-06-01

    Full Text Available In this study the effects of an aqueous suspension of a commercial preparation of the mushroom Coprinus comatus on oxidative stress induced in rats by alloxane and carbon tetrachloride was examined. The effects were estimated from changes in the biochemical parameters (xanthine oxidase, glutathione peroxidase and catalase activity, reduced glutathione content, and extent of lipid peroxidation of liver homogenate as well as histological changes in the liver of the rats treated with alloxane and carbon tetrachloride. Two screening doses of alloxane sufficient to induce diabetes in rats did not have any significant effect on the examined biochemical parameters of liver homogenate or on the cytoarchitectonics of liver cross-sections. Treatment with carbon tetrachloride resulted in a significant increase in the intensity of lipid peroxidation and peroxydasis activity, as well as with decrease in catalase activity. Certain changes in liver cross sections were detected, such is lymphocyte infiltration of dilated sinusoid capillaries. Administration of Coprinus comatus suspension thus showed antioxidative potential, evidenced by an increase of antioxidative status of liver homogenate and prevention of histological changes in liver cross sections.

  20. Anti-oxidative activity of an aqueous suspension of commercial preparation of the mushroom Coprinus comatus.

    Science.gov (United States)

    Popović, Mira; Vukmirović, Sasa; Stilinović, Nebojsa; Capo, Ivan; Jakovljević, Vida

    2010-06-24

    In this study the effects of an aqueous suspension of a commercial preparation of the mushroom Coprinus comatus on oxidative stress induced in rats by alloxane and carbon tetrachloride was examined. The effects were estimated from changes in the biochemical parameters (xanthine oxidase, glutathione peroxidase and catalase activity, reduced glutathione content, and extent of lipid peroxidation) of liver homogenate as well as histological changes in the liver of the rats treated with alloxane and carbon tetrachloride. Two screening doses of alloxane sufficient to induce diabetes in rats did not have any significant effect on the examined biochemical parameters of liver homogenate or on the cytoarchitectonics of liver cross-sections. Treatment with carbon tetrachloride resulted in a significant increase in the intensity of lipid peroxidation and peroxydasis activity, as well as with decrease in catalase activity. Certain changes in liver cross sections were detected, such is lymphocyte infiltration of dilated sinusoid capillaries. Administration of Coprinus comatus suspension thus showed antioxidative potential, evidenced by an increase of antioxidative status of liver homogenate and prevention of histological changes in liver cross sections.

  1. Secrets Law

    Directory of Open Access Journals (Sweden)

    Luz Helena Guamanzara Torres

    2013-01-01

    Full Text Available This paper provides a review of the book The Law of Secrets, of the author Juan Carlos Martínez-Villalba Riofrío studying the secrets and how law does protect. To this end, the author has analyzed the general theory of secrecy, secrets and methodology, its overall rating, essential elements and their different legal dimensions, the secret as a subjective right. It also establishes that professional secrecy is protected by constitutional principles such as the right to privacy.

  2. Efficient GFP expression in the mushrooms Agaricus bisporus and Coprinus cinereus requires introns.

    Science.gov (United States)

    Burns, C; Gregory, K E; Kirby, M; Cheung, M K; Riquelme, M; Elliott, T J; Challen, M P; Bailey, A; Foster, G D

    2005-03-01

    We have developed a "Molecular Toolkit" comprising interchangeable promoters and marker genes to facilitate transformation of homobasidiomycete mushrooms. We describe the evaluation of a range of promoters in the homobasidiomycetes Agaricus bisporus and Coprinus cinereus using green fluorescent protein (GFP) as a reporter gene; the C. cinereus trp1 promoter and A. bisporus trp2 and gpdII promoters proving successful in driving expression in C. cinereus, with the gpdII promoter also functioning in A. bisporus. Our investigations demonstrate that a prerequisite for GFP expression in C. cinereus and A. bisporus is the presence of an intron. This is the first reported expression of GFP in either C. cinereus or A. bisporus.

  3. Comparison of hypoglycemic activity of trace elements absorbed in fermented mushroom of Coprinus comatus.

    Science.gov (United States)

    Lv, Yingtao; Han, Linna; Yuan, Chao; Guo, Jianyou

    2009-11-01

    The effect of fermented mushroom of Coprinus comatus rich in trace elements, including vanadium, chromium, zinc, magnesium, copper, iron, and nickel, on glycemic metabolism was studied in this paper. Alloxan-induced hyperglycemic mice were used in the study. The blood glucose, glycohemoglobin, and glycogen synthesis of the mice were analyzed, respectively. At the same time, the gluconeogenesis of the normal mice was also determined. After the mice were administered (ig) with C. comatus rich in vanadium (CCRV), the blood glucose and the glycohemoglobin of alloxan-induced hyperglycemic mice decreased (p < 0.05, p < 0.01), glycogen synthesis of alloxan-induced hyperglycemic mice elevated (p < 0.01), the gluconeogenesis of the normal mice was inhibited (p < 0.01), and the sugar tolerance of the normal mice was improved. However, the same result did not occur in other groups. Vanadium at lower doses in combination with C. comatus induced significant effect on glycemic metabolism in mice.

  4. Morphometric analysis of cell size patterning involved in gravitropic curvature of the stipe of coprinus cinereus

    Science.gov (United States)

    Greening, J. P.; Moore, D.

    During gravitropic bending of the stipe of Coprinus cinereus the majority of elongation occurred in the apical region of the lower surface of the stipe, although some elongation was seen throughout the stipe. The final rate of elongation was similar at both the upper and lower stipe surfaces but the lower surface achieved this rate first (close to the reaction time 25 min /2/), whilst the upper surface of the stipe only attained its final elongation rate after a period of acceleration of 150 min. Detailed morphometric analysis of cell size patterning in transverse sections revealed no significant differences in cross sectional area, spatial or proportional distribution of different cell types between the upper and lower regions of the gravitropic bend. Measurements of longitudinal cell size revealed significant differences in compartment size between the lower and upper region. Hyphal compartments of lower regions of the bend were on average four to five times longer than those of the upper region.

  5. A comparison of methods for successful triggering of gene silencing in Coprinus cinereus.

    Science.gov (United States)

    Heneghan, Mary N; Costa, Ana M S B; Challen, Michael P; Mills, Peter R; Bailey, Andy; Foster, Gary D

    2007-03-01

    Post-transcriptional gene-silencing methods (PTGS), including RNAi, are becoming increasingly pervasive in functional genomics. To advance analysis of the recently sequenced Coprinus cinereus genome, a high throughput gene silencing method is essential. We have exploited the GFP reporter gene to evaluate and quantify efficacy of three different silencing strategies. Modular constructs that encompassed antisense, untranslatable sense, and RNAi-mediating hairpin sequences, were transformed into a GFP-expressing host strain. Transformants exhibiting strong downregulation and partial suppression of GFP were recovered with all three constructs. Analyses of protein and transcriptional nucleic acids revealed that the antisense and hairpin sequences yielded similar levels of GFP suppression, and were both more efficient than untranslatable sense sequences. Our antisense vectors will expedite functional characterisation of C. cinereus and the modular nature of the constructs should permit exploitation of directional cDNA libraries for high throughput screening.

  6. Hypoglycemic activity of fermented mushroom of Coprinus comatus rich in vanadium.

    Science.gov (United States)

    Han, Chunchao; Yuan, Junhua; Wang, Yingzi; Li, Lingjun

    2006-01-01

    The hypoglycemic activity of fermented mushroom of Coprinus comatus rich in vanadium (CCRV) was studied in this paper. Alloxan and adrenalin induced hyperglycemic mice were used in the study. The blood glucose and the HbA1c of the mice were analyzed, respectively. At the same time, the sugar tolerance of the normal mice was also determined. After the mice were administered (ig) with CCRV, the blood glucose and the HbA1c of alloxan-induced hyperglycemic mice decreased (p < 0.05, p < 0.01), ascension of blood glucose induced by adrenalin was inhibited (p < 0.01) and the sugar tolerance of the normal mice was improved. Also, the body weight of the alloxan-induced hyperglycemic mice was increased gradually. In the fermented mushroom of C. comatus, vanadium at lower doses in combination with C. comatus, induced significant decreases of the blood glucose and HbA1c levels in hyperglycemic mice.

  7. [Production of ligninase by co-fermentation of Coprinus comatus and Trichoderma reesei].

    Science.gov (United States)

    Ge, Chunmei; Xu, Juanjuan; Sun, Qinying; Zhang, Jie; Cai, Jingmin; Pan, Renrui

    2009-12-01

    In order to enhance the utilization efficiency, reduce the environmental pollution of traditional chemical treatment and the agriculture waste incineration; we studied the ligninase production by Coprinus comatus, Aspergillus niger and Trichoderma reesei through the plate screening. The results showed that C. comatus mixed culture with T. reesei have a good compatibility and higher yields of Laccase. On the basis of this pre-experiment, we studied the optimal conditions of mixed culture for enzyme production. Under the optimal conditions: the inoculation proportion of C. comatus and T. reesei (5:2), the interval of time (12 h), the temperature 260C, the shake rotation speed 150 r/min, fermented for 3 days, the Laccase activity reached 3267.1 U/mL, increased by 106% contrasted with single culture of C. commatus.

  8. Treatment of oil refinery wastewater using crude Coprinus cinereus peroxidase and hydrogen peroxide

    Energy Technology Data Exchange (ETDEWEB)

    Ikehata, K.; Buchanan, I. D.; Smith, D. W. [University of Alberta, Dept. of Civil and Environmental Engineering, Edmonton, AB (Canada)

    2003-11-01

    Enzymatic treatment of oil refinery wastewater was investigated using crude peroxidase derived from the fungus Coprinus cinereus (CIP), and hydrogen peroxide. Further objectives were to investigate the effects of residual organic compounds in the crude enzyme, and compare the performance of CIP to those of purified horseradish peroxidase (HRP) and Arthromyces ramosus peroxidase (ARP) in the treatment of a strong refinery wastewater. Phenols in the wastewater were converted to coloured polymeric products and then removed by coagulation with alum. As a result of the enzymatic treatment and alum coagulation of the wastewater containing 6.4 mM total phenol, the chemical oxygen demand and the 5-d biochemical oxygen demand were reduced by 52 per cent and 58 per cent, respectively. Reduction of the oxygen demands notwithstanding, the dissolved organic materials in the crude CIP were not affected by either of these processes and tended to remain in the treated wastewater. 31 refs., 1 tab., 7 figs.

  9. Determination of some biological activities of Tricholoma terreum and Coprinus micaceus

    Directory of Open Access Journals (Sweden)

    Hasan Akgül

    2016-10-01

    Full Text Available The use of mushrooms as food also as medicine is increasing day by day. In this study, an edible mushrooms - Tricholoma terreum (Schaeff. P. Kumm. and Coprinus micaceus (Bull. Fr. – were used as materials. This study was aimed to determine the antioxidant and DNA protective activity of ethanol extract of mushrooms and determine and compare total antioxidant, oxidative stress index with oxidative levels and Fe, Mg, Zn, Cu, Na and Ca contents. The antioxidant activity of mushrooms were analyzed with using DPPH and total antioxidant and oxidative levels were analyzed with using Rel Assay Diagnostics kits. DNA protective activity was determined by using the supercoiled pBR322 DNA. Furthermore, element contents of the mushrooms were determined with wet decomposition method by using atomic absorption spectrophotometry. As a result of the study, antioxidant and DNA protective activities of ethanol extracts of both mushroom species were not showed any notable effect and total oxidant value was determined very high.

  10. Coprinus comatus (shaggy cap) is a potential source of aeroallergen that may provoke atopic dermatitis.

    Science.gov (United States)

    Fischer, B; Yawalkar, N; Brander, K A; Pichler, W J; Helbling, A

    1999-10-01

    Basidiospores are universal components in the air and established as important causes of respiratory allergies. Recent reports indicate that aeroallergens may aggravate eczematous skin lesions in subjects with atopic dermatitis (AD). The aim of the study was to investigate whether spores of Coprinus comatus, a species of basidiomycetes, may elicit delayed-type skin reactions in subjects with an atopic predilection, especially dermatitis. Sixty-six study subjects were categorized in groups having AD or respiratory allergy with regard to the skin prick test (SPT) reactivity to C comatus extract. Twenty nonatopic individuals served as control subjects. Atopy patch tests (APTs) were performed with extract of C comatus spore containing tissue at a concentration of 1. 35 mg of protein per gram of petrolatum (Vaseline) and C comatus cap at a concentration of approximately 5 mg of protein per gram of petroleum jelly. APT reactions were evaluated after 48 and 72 hours. Of the subjects with AD completing the study, 12 (32%) of 38 showed a positive APT reaction, with 8 (57%) also having a positive SPT response to C comatus. Only 1 (9%) of 11 subjects with asthma had a positive SPT and APT response to C comatus. No positive test reaction was observed in the nonatopic control subjects or in subjects with respiratory allergy and negative SPT responses to C comatus. Our results demonstrate that C comatus can induce delayed-type reactions in atopic individuals, particularly in those with AD. Because spores of Coprinus species are ubiquitous, basidiomycetes have to be considered as possible aeroallergens when investigating causes of eczematous skin lesions in AD.

  11. Mercury bio-extraction by fungus Coprinus comatus: a possible bioindicator and mycoremediator of polluted soils?

    Science.gov (United States)

    Falandysz, Jerzy

    2016-04-01

    The Shaggy Ink Cap (Coprinus comatus), which is a common in wild in northern hemisphere was examined in field for potential to be used as possible bio-extractor of Hg from polluted grounds but also as possible bioindicator of urban soils (roadside, barren lands, lawns) pollution with Hg. The contents of Hg in caps and stipes of C. comatus from the grounds examined in this study correlated positively with the levels of soil contamination. Analysis of sets of data available worldwide on Hg in C. comatus and soils beneath-fruiting bodies showed on a positive correlation between degree of soil and mushroom contamination. Hence, C. comatus could be considered as a sensitive species and with bioindication and bioremediation potency for soils polluted with Hg in further studies. Young-fruiting bodies of C. comatus are edible and considered excellent if consumed soon after pick-up. Eating them when foraged from the urban places can provide to a consumer Hg at relatively high dose, while unresolved question is absorption rate of Hg compounds contained in ingested mushroom meal.

  12. Two classes of homeodomain proteins specify the multiple a mating types of the mushroom Coprinus cinereus.

    Science.gov (United States)

    Kües, U; Asante-Owusu, R N; Mutasa, E S; Tymon, A M; Pardo, E H; O'Shea, S F; Göttgens, B; Casselton, L A

    1994-01-01

    The A mating type locus of the mushroom Coprinus cinereus regulates essential steps in sexual development. The locus is complex and contains several functionally redundant, multiallelic genes that encode putative transcription factors. Here, we compare four genes from an A locus designated A42. Overall, the DNA sequences are very different (approximately 50% homology), but two classes of genes can be distinguished on the basis of a conserved homeodomain motif in their predicted proteins (HD1 and HD2). Development is postulated to be triggered by an HD1 and an HD2 gene from different A loci. Thus, proteins encoded by genes of the same locus must be distinguished from those encoded by another locus. Individual proteins of both classes recognize each other using the region N-terminal to the homeodomain. These N-terminal specificity regions (COP1 and COP2) are predicted to be helical and are potential dimerization interfaces. The amino acid composition of the C-terminal regions of HD1 proteins suggests a role in activation, and gene truncations indicate that this region is essential for function in vivo. A corresponding C-terminal region in HD2 proteins can be dispensed with in vivo. We will discuss these predicted structural features of the C. cinereus A proteins, their proposed interactions following a compatible cell fusion, and their similarities to the a1 and alpha 2 mating type proteins of the yeast Saccharomyces cerevisiae. PMID:7994179

  13. Some morphological and physiological aspects and cultivation of Coprinus comatus (O. F. Mull. Gray

    Directory of Open Access Journals (Sweden)

    Petcharat, V.

    2007-03-01

    Full Text Available Some morphological and physiological aspects and cultivation of Coprinus comatus (O.F. Mull. Gray were investigated. Malt extract agar turned out to be the best in supporting the mycelial growth of C.comatus. Mannose and maltose were the best carbon sources in supporting mycelial growth. C. comatus utilized peptone and NH4NO3 better than other nitrogen sources. The optimum temperature and pH on maltextract agar were 25oC and 6, respectively. Light retarded mycelial growth of C. comatus. For cultivation, the method of growing mushroom in autoclavable plastic bags was applied. Three different combinations of agricultural products were used for growing C. comatus. The combination ofpararubber sawdust : kapok waste : boiled sorghum seeds (3:3:1 by volume supported higher yields of basidiocarps. Time required for full colonization of the mycelia on 500 gm substrate at room temperature(28-30oC was 20.3 days. After casing with the mixed soil (loam soil : rice husk : cow manure, 2:2:1 the cultivating bags were incubated in the growth chamber at 20oC and 65% relative humidity. Fructificationbegan after 20.6 days of watering, and the average yield obtained was 165.6 g/bag.

  14. Expression of flap endonuclease-1 during meiosis in a basidiomycete, Coprinus cinereus.

    Science.gov (United States)

    Yamaguchi, Taiki; Namekawa, Satoshi H; Hamada, Fumika N; Kasai, Nobuyuki; Nara, Takayuki; Watanabe, Kei; Iwabata, Kazuki; Ishizaki, Takashi; Ishii, Satomi; Koshiyama, Akiyo; Inagaki, Sachiyo; Kimura, Seisuke; Sakaguchi, Kengo

    2004-05-01

    In the basidiomycete Coprinus cinereus (C. cinereus), which shows a highly synchronous meiotic cell cycle, the meiotic prophase I cells demonstrate flap endonuclease-1 activity. To investigate its role during meiosis, we isolated a C. cinereus cDNA homolog of flap endonuclease-1 (CcFEN-1), 1377bp in length with the open reading frame (ORF) encoding a predicted molecular mass of 51 kDa. At amino-acid residues Glu276-Pro345, a specific inserted sequence composed of 70 amino acids rich in polar forms was found to exist, without sequence identity to other eukaryotic FEN-1 or the polar amino acid rich sequences found in C. cinereus PCNA and C. cinereus DNA ligase IV, although the lengths and percentages of polar amino acids were similar. Northern hybridization analysis indicated CcFEN-1 to be expressed not only in the pre-meiotic S phase but also in meiotic prophase I. The roles of CcFEN-1 during meiosis are discussed.

  15. The comparision of Coprinus cinereus peroxidase enzyme and TiO2 catalyst for phenol removal.

    Science.gov (United States)

    Sarkhanpour, Reza; Tavakoli, Omid; Sarrafzadeh, Mohammad Hossein; Kariminia, Hamid Reza

    2013-01-01

    This article investigates phenol removal from an aqueous solution by using enzymatic and photocatalytic methods and the efficiency of these methods has been compared. In enzymatic and photocatalytic methods, Coprinus cinereus, peroxidase enzyme and commercial TiO(2) powders (Degussa P-25) in aqueous suspension were used, respectively, in ambient temperature. The effects of different operating parameters such as duration of process, catalyst dosage or enzyme concentration, pH of the solution, initial phenol concentration and H(2)O(2) concentration on both processes were examined. In enzymatic method, efficiency of degradation reached 100% within 5 min, while in the photocatalytic method, the efficiency of degradation reached approximately 70% within 60 min. In photocatalytic method, there is an optimum concentration for catalyst dosage (near 2.0 g/L) to gain 80% efficiency, while in the enzymatic method, increasing the amount of enzyme could lead to an increase in the efficiency up to 100%. Moreover, the optimum pH in enzymatic and photocatalytic methods stood at 8.0 and 7.0, respectively. In both methods, the addition of different amounts of H(2)O(2) increased the degradation efficiency to 100%.

  16. Two X family DNA polymerases, lambda and mu, in meiotic tissues of the basidiomycete, Coprinus cinereus.

    Science.gov (United States)

    Sakamoto, Aiko; Iwabata, Kazuki; Koshiyama, Akiyo; Sugawara, Hiroko; Yanai, Takuro; Kanai, Yoshihiro; Takeuchi, Ryo; Daikuhara, Yoko; Takakusagi, Yoichi; Sakaguchi, Kengo

    2007-12-01

    The X family DNA polymerases lambda (CcPollambda) and mu (CcPolmu) were shown to be expressed during meiotic prophase in the basidiomycete, Coprinus cinereus. These two polymerases are the only members of the X family in the C. cinereus genome. The open reading frame of CcPollambda encoded a predicted product of 800 amino acid residues and that of CcPolmicro of 621 amino acid residues. Both CcPollambda and CcPolmicro required Mn(2+) ions for activity, and both were strongly inhibited by dideoxythymidine triphosphate. Unlike their mammalian counterparts, CcPollambda and CcPolmicro had no terminal deoxynucleotidyl transferase activity. Immunostaining analysis revealed that CcPollambda was present at meiotic prophase nuclei in zygotene and pachytene cells, which is the period when homologous chromosomes pair and recombine. CcPolmicro was present in a slightly wider range of cell stages, zygotene to diplotene. In analyses using D-loop recombination intermediate substrates, we found that both CcPollambda and CcPolmicro could promote primer extension of an invading strand in a D-loop structure. Moreover, both polymerases could fully extend the primer in the D-loop substrate, suggesting that D-loop extension is an activity intrinsic to CcPollambda and CcPolmicro. Based on these data, we discuss the possible roles of these polymerases in meiosis.

  17. An mre11 mutant of Coprinus cinereus has defects in meiotic chromosome pairing, condensation and synapsis.

    Science.gov (United States)

    Gerecke, E E; Zolan, M E

    2000-01-01

    The rad11 gene of the basidiomycete Coprinus cinereus is required for the completion of meiosis and for survival after gamma irradiation. We have cloned the rad11 gene and shown that it is a homolog of MRE11, a gene required for meiosis and DNA repair in numerous organisms. The expression of C. cinereus mre11 is induced during prophase I of meiosis and following gamma irradiation. The gene encodes a predicted polypeptide of 731 amino acids, and the mre11-1 (rad11-1) mutation is a single base pair change that results in a stop codon after amino acid 315. The mre11-1 mutant shows enhanced sensitivity to ionizing radiation, but no enhanced sensitivity to UV radiation. It shows a delay in fruitbody formation and a reduction in the number of mushrooms formed per dikaryon. The mre11-1 mutant also has several meiotic defects. Pachytene chromatin condensation is disrupted, and although some meiotic cells appear to achieve metaphase I condensation, no further meiotic progression is observed. The mre11-1 mutant also fails to undergo proper chromosome synapsis; neither axial elements nor mature synaptonemal complexes are complete. Finally, meiotic homolog pairing is reduced in the mre11-1 mutant. Thus, in C. cinereus, Mre11 is required for meiotic DNA metabolism. PMID:10757758

  18. Production and Characterization of Radiation-Sensitive Meiotic Mutants of Coprinus Cinereus

    Science.gov (United States)

    Zolan, M. E.; Tremel, C. J.; Pukkila, P. J.

    1988-01-01

    We have isolated four γ-ray-sensitive mutants of the basidiomycete Coprinus cinereus. When homozygous, two of these (rad 3-1 and rad 9-1) produce fruiting bodies with very few viable basidiospores, the products of meiosis in this organism. A less radiation-sensitive allele of RAD 3, rad 3-2, causes no apparent meiotic defect in homozygous strains. Quantitative measurements of oidial survival of rad 3-1;rad 9-1 double mutants compared to the single mutants indicated that rad 3-1 and rad 9-1 mutants are defective in the same DNA repair pathway. In the few viable basidiospores that are produced by these two strains, essentially normal levels of meiotic recombination can be detected. None of the mutants exhibits increased sensitivity to UV radiation. Cytological examination of meiotic chromosomes from mutant and wild-type fruiting bodies showed that rad 3-1 homozygous strains fail to condense and pair homologous chromosomes during prophase I. Although rad 9-1 strains are successful at chromosome pairing, meiosis is usually not completed in these mutants. PMID:3197952

  19. Improving the synthesis of phenolic polymer using Coprinus cinereus peroxidase mutant Phe230Ala.

    Science.gov (United States)

    Kim, Su Jin; Joo, Jeong Chan; Song, Bong Keun; Yoo, Young Je; Kim, Yong Hwan

    2016-06-01

    The F230A mutant of Coprinus cinereus peroxidase (CiP), which has a high stability against radical-inactivation, was previously reported. In the present study, the radical-robust F230A mutant was applied to the oxidative polymerization of phenol. The F230A mutant exhibited better polymerization activities than the wild-type CiP in the presence of water-miscible alcohols i.e., methanol, ethanol, and isopropanol despite its lower stability against alcohols. In particular, the F230A mutant showed a higher consumption of phenol (40%) and yielded phenolic polymer of larger molecular weight (8850Da) in a 50% (v/v) isopropanol-buffer mixture compared with the wild-type CiP (2% and 1519Da, respectively). In addition, the wild-type CiP and F230A mutant had no significant differences in enzyme inactivation by physical adsorption on the polymeric products or by heat incubation, and showed comparable kinetic parameters. These results indicate that high radical stability of the F230A mutant and improved solubility of phenolic polymers in alcohol-water cosolvent systems may synergistically contribute to the production of the high molecular weight phenolic polymer. Copyright © 2016 Elsevier Inc. All rights reserved.

  20. Purification and characterization of a novel laccase from Coprinus cinereus and decolorization of different chemically dyes.

    Science.gov (United States)

    Lin, Yaqiu; Zhang, Zhen; Tian, Yongsheng; Zhao, Wei; Zhu, Bo; Xu, Zhisheng; Peng, Rihe; Yao, Quanhong

    2013-02-01

    Laccase is a blue copper oxidase with multiple copper ions and widely distributed in higher plant and fungi. To date, numerous fungal laccases have been reported by many researchers. In present work, a new laccase gene, named CcLCC5I, from Coprinus cinereus was synthesized chemically according to the yeast bias codon and integrated into Pichia pastoris GS115 genome by electroporation. SDS-PAGE analysis showed that the recombinant laccase has a molecular mass of approximately 56.8 kDa. Its biochemical properties was carried out using substrate 2-2(')-azino-bis(3-ethylbenzothiazoline-6-sulfonate) (ABTS). It was showed that the optimum pH and temperature of the laccase is 3.0 and 55 °C, respectively. Except for copper ions, most metal ions inhibited the laccase activity at a high concentration about 10 mM. Sodium sulfite can also highly inhibit laccase activity whereas EDTA had no inhibitory effect on the laccase activity. The CcLCC5I have high ability to decolor not only azo but also aryl methane dyes. The recombinant laccase decolored 44.6 % orange G, 54.8 % Crystal Violet, and 87.2 % Malachite green at about 2.6 h. The novel laccase may be a good candidate for breeding engineering strains used in the treatment of industrial effluent containing azo and aryl methane dyes.

  1. Immobilized Coprinus plicatilis Biodegradation of Fluorene in Two Different Packed-Bed Reactors.

    Science.gov (United States)

    Akdogan, Hatice Ardag

    2015-01-01

    The biodegradation of fluorene by immobilized Coprinus plicatilis was studied in pinewood and foam glass bead-packed reactors. The reactors were operated in a sequencing batch system. Removal efficiency increased over time and elevated influent fluorene concentration (85 mg/L) was removed 100% in 24-30 h batch cycles. Increased laccase activity was detected with the introduction of the compounds, and optimum activity corresponded to optimum removal periods. Significantly higher laccase activity (16.7-19 U/L) was detected in the glass bead-packed reactor compared to the pinewood-packed reactor (0.2-5 U/L). The presence of Mn2+ ions in the wood material possibly caused elevated manganese peroxidase activity (0.3-5.8 U/L) compared to low to negligible activity in the glass bead reactor. Reactor performances are discussed in relation to sequencing batch operation and nutrient requirements necessary to induce and sustain fungal enzyme activity in inert-like organic material packed systems. Biodegradation metabolites were detected in samples via GC/MS.

  2. Coprinus cinereus Mer3 is required for synaptonemal complex formation during meiosis.

    Science.gov (United States)

    Sugawara, Hiroko; Iwabata, Kazuki; Koshiyama, Akiyo; Yanai, Takuro; Daikuhara, Yoko; Namekawa, Satoshi H; Hamada, Fumika N; Sakaguchi, Kengo

    2009-02-01

    Mer3 is an evolutionarily conserved DNA helicase that has crucial roles in meiotic recombination and crossover formation. We have identified the MER3 homolog in Coprinus cinereus (Ccmer3) and show that it is expressed in zygotene and pachytene meiocytes. Immunostaining analysis indicated that CcMer3 was localized on chromosomes at zygotene and pachytene and CcMer3 foci were more frequent on paired than unpaired chromosomes. We generated a C. cinereus mer3 mutant (#1) and found that it showed abnormal meiosis progression and underwent apoptosis after prophase I. Basidiospore production in #1 was reduced to 0.8% of the wild-type level; the spores showed slower germination at 25 degrees C but were similar to the wild type at 37 degrees C. Electron microscopic analysis of chromosome spreads revealed that axial elements were formed in the mutant but that synapsis was defective, resulting in a reduction in spore production. Our results demonstrate that CcMer3 is required for synaptonemal complex formation after axial elements align and is thus essential for homologous synapsis.

  3. Antitumor and immunomodulatory activity of a polysaccharide from fungus Coprinus comatus (Mull.:Fr.) Gray.

    Science.gov (United States)

    Jiang, Xiao-Gang; Lian, Ming-Xue; Han, Yan; Lv, She-Min

    2013-07-01

    In our study, a water-soluble polysaccharide (CCPa-1) was successfully purified from the fruiting bodies of Coprinus comatus by DEAE-cellulose and Sepharose CL-6B column chromatography. The molecular weight was evaluated to be 53.6 kDa as determined by high performance size exclusion chromatography (HPSEC). Sugar composition analysis revealed that CCPa-1 consisted primarily of galactose, glucose and arabinose in a molar ratio of 6.6:1.2:2.2. CCPa-1 could not only inhibit the growth of sarcoma 180 (S180) tumor transplanted in mice, but also increase the relative spleen/thymus indexes and body weight of tumor bearing mice. Moreover, Con A- or LPS-induced splenocyte proliferation was also enhanced after CCPa-1 administration in tumor-bearing mice. Furthermore, CCPa-1 significantly enhanced the Con A- or LPS-induced splenocyte proliferation and increased the production of TNF-α and IL-2. All the data demonstrated that CCPa-1 had a potential application as natural antitumor agent with immunomodulatory activity. Copyright © 2013 Elsevier B.V. All rights reserved.

  4. Structural investigation of water-soluble polysaccharides extracted from the fruit bodies of Coprinus comatus.

    Science.gov (United States)

    Li, Bo; Dobruchowska, Justyna M; Gerwig, Gerrit J; Dijkhuizen, Lubbert; Kamerling, Johannis P

    2013-01-02

    Water-soluble polysaccharide material, extracted from the stipes of the fruit bodies of Coprinus comatus by hot water, was fractionated by sequential weak anion-exchange and size-exclusion chromatography. The relevant fractions were subjected to structural analysis, including (d/l) monosaccharide/methylation analysis and 1D/2D NMR spectroscopy. Besides the disaccharide α,α-trehalose [α-D-Glcp-(1↔1)-α-D-Glcp], high-molecular-mass α-D-glucans (the most abundant component) consisting of [→4)-α-D-Glcp-(1→](n) backbones with ~10% branching at C-6 by terminal α-D-Glcp-(1→6)- or α-D-Glcp-(1→6)-α-D-Glcp-(1→6)- units, lower-molecular-mass linear β-D-glucans consisting of [→6)-β-D-Glcp-(1→](m) sequences, and a lower-molecular-mass pentasaccharide-repeating α-L-fuco-α-D-galactan, {→6)-α-D-Galp-(1→6)-[α-L-Fucp-(1→2)-]α-D-Galp-(1→6)-α-D-Galp-(1→6)-α-D-Galp-(1→}(p), were found to be present. Copyright © 2012 Elsevier Ltd. All rights reserved.

  5. Turnover capacity of Coprinus cinereus peroxidase for phenol and monosubstituted phenols

    Energy Technology Data Exchange (ETDEWEB)

    Aitken, M.D.; Heck, P.E. [Univ. of North Carolina, Chapel Hill, NC (United States)

    1998-05-01

    Coprinus cinereus peroxidase (CIP) and other peroxidases are susceptible to mechanism-based inactivation during the oxidation of phenolic substrates. The turnover capacity of CIP was quantified for phenol and 11 monosubstituted phenols under conditions in which enzyme inactivation by mechanisms involving hydrogen peroxide alone were minimized. Turnover capacities varied by nearly 2 orders of magnitude, depending on the substituent. On a mass basis, the enzyme consumption corresponding to the lowest turnover capacities is considerable and may influence the economic feasibility of proposed industrial applications of peroxidases. Within a range of substituent electronegativity values, molar turnover capacities correlated well (r{sup 2} = 0.89) with substituent effects quantified by radical {sigma} values and semiquantitatively with homolytic O-H bond dissociation energies of the phenolic substrates, suggesting that phenoxyl radical intermediates are probably involved in the suicide inactivation of CIP. The correlation range in each case did not include phenols with highly electron-withdrawing (nitro and cyano) substituents because they are not oxidized by CIP, nor phenols with highly electron-donating (hydroxy and amino) substituents because they led to virtually complete inactivation of the enzyme with minimal substrate removal.

  6. Molecular Analysis of the Coprinus Cinereus Mating Type a Factor Demonstrates an Unexpectedly Complex Structure

    Science.gov (United States)

    May, G.; Chevanton, L. L.; Pukkila, P. J.

    1991-01-01

    We report here the molecular cloning of the A43 mating type factor from Coprinus cinereus, a basidiomycetous fungus. Our molecular analyses revealed an unexpected source of variation in the A factor. Though genetic studies have demonstrated that A has two subunits, α and β, we located three nonoverlapping fragments in the A43 region that have A factor function following DNA-mediated transformation. The three fragments demonstrate no similarity to one another as judged by restriction enzyme maps and by hybridization on Southern blots. We conclude that the A43 factor is composed of at least three subunits. When strains carrying different A factors are examined by hybridization to the cloned subunits, extensive polymorphism is seen. Both intensity of hybridization and restriction fragment lengths vary between strains. Some strains fail to show any hybridization to a probe. In contrast, other strains from widely separated geographic locations apparently share very similar subunits. From comparative restriction enzyme mapping of A43 and a mutated A43 factor, we inferred that a 12-kb deletion in the A factor was responsible for the constitutive, dominant phenotype of the mutated A factor. The results of transformation experiments support an activator model for the activity of the A factor in regulating the A pathway. PMID:1678725

  7. Correlation of Genetic and Physical Maps at the a Mating-Type Locus of Coprinus Cinereus

    Science.gov (United States)

    Lukens, L.; Yicun, H.; May, G.

    1996-01-01

    The A mating type locus of Coprinus cinereus is remarkable for its extreme diversity, with over 100 different alleles in natural populations. Classical genetic studies have demonstrated that this hypervariability arises in part from recombination between two subloci of A, alpha and beta, although more recent population genetic data have indicated a third segregating sublocus. In this study, we characterized the molecular basis by which recombination generates nonparental A mating types. We mapped the frequency and location of all recombination events in two crosses and correlated the genetic and physical maps of A. We found that all recombination events were located in 6 kb of noncoding DNA between the alpha and beta subloci and that the rate of recombination in this noncoding region matched that generally observed for this genome. No recombination within gene clusters or within coding regions was observed, and the two alpha and beta subloci described in genetic analyses correlated with the previously characterized alpha and beta gene clusters. We propose that pairs of genes constitute both the sex determining and the hereditary unit of A. PMID:8978036

  8. The culinary-medicinal mushroom Coprinus comatus as a natural antiandrogenic modulator.

    Science.gov (United States)

    Dotan, Nesly; Wasser, Solomon P; Mahajna, Jamal

    2011-06-01

    Prostate cancer is the most common cancer diagnosed in men. Chemotherapy, androgen ablation, and androgen antagonist treatments have proven to have significant effects in the early stages of prostate cancer, whereas advanced prostate cancer is resilient to such treatments. The androgen receptor (AR), a ligand-dependent transcription factor, is the major drug target of prostate cancer therapy. Transition to the androgen-independent stage involves the activation of signaling pathways, AR gene mutations, and other mechanisms. Higher basidiomycetes mushrooms have been used since ancient times in folk medicine to treat a diversity of diseases, including cancer. The present study evaluates the antiandrogenic activity of different Coprinus comatus strains in their ability to interfere with AR function. The authors found that the most active extract was C comatus strain 734 extracted with hexane (CC734-H). This extract was able to (1) inhibit AR-mediated reporter activity, (2) inhibit the proliferation and viability of the LNCaP cell line, and (3) inhibit the colony formation of the LNCaP cell line, in comparison to the DU-145, PC-3, and MDA-Kb2 cells. In addition, CC734-H was able to reduce AR levels and prostate-specific antigen gene expression in the LNCaP-treated cell line. This study illustrates the potential of the C comatus mushroom as a natural antiandrogenic modulator that could serve in the treatment of prostatic diseases.

  9. Coprinus comatus damages nematode cuticles mechanically with spiny balls and produces potent toxins to immobilize nematodes.

    Science.gov (United States)

    Luo, Hong; Liu, Yajun; Fang, Lin; Li, Xuan; Tang, Ninghua; Zhang, Keqin

    2007-06-01

    We reported recently a unique fungal structure, called the spiny ball, on the vegetative hyphae of Coprinus comatus (O. F. Müll.:Fr.) Pers. Although some observations regarding the role of this structure were presented, its function remained largely unknown. In this study, we showed that purified (isolated and washed) spiny balls could immobilize and kill the free-living nematode Panagrellus redivivus Goodey highly efficiently. Scanning electron microscopy studies illustrated that the spiny structure damaged the nematode cuticle, suggesting the presence of a mechanical force during the process of nematode immobilization. Severe injuries on nematode cuticles caused the leakage of inner materials of the nematodes. When these structures were ground in liquid nitrogen, their killing efficacy against nematodes was lost, indicating that the shape and the complete structure of the spiny balls are indispensable for their function. However, extraction with organic solvents never lowered their activity against P. redivivus, and the extracts showed no obvious effect on the nematode. We also investigated whether C. comatus was able to produce toxins which would aid in the immobilization of nematodes. In total, we identified seven toxins from C. comatus that showed activity to immobilize the nematodes P. redivivus and Meloidogyne incognita (Kofoid et White) Chitwood. The chemical structures of these toxins were identified with nuclear magnetic resonance, mass spectrometry, infrared, and UV spectrum analysis. Two compounds were found to be novel. The toxins found in C. comatus are O-containing heterocyclic compounds.

  10. [Purification and activities of an alkaline protein from mushroom Coprinus comatus].

    Science.gov (United States)

    Wu, Liping; Wu, Zujian; Lin, Qiying; Xie, Lianhui

    2003-12-01

    An Alkaline protein, y3, can be purified from the fruiting bodies of mushroom Coprinus comatus by means of CM-sepharose FF ion-exchange column chromatography and Superdex 75 High Resolution molecular sieve chromatography. The protein has a molecular weight of about 14.4kD by SDS-PAGA. Some activities of y3 have been detected, and the result is the following: the inhibition rate against Tobacco Mosaic Virus is 83.0% when the concentration of y3 is 12.5 microg/ mL. y3 is able to agglutinate rabbit and human erythrocytes at the concentration of 1.562 microg/mL and 0.781 microg/mL. Using an assay system based on mach cancer cell line MGC-803, y3 was studied for its inhibitory ability against celles multiplication, and the IC50 is 12 microg/mL. The N-terminal sequence is NRDVAACARFIDDFCDTLTP, which has no homology with other sequences in Genbank.

  11. Coprinus comatus: A basidiomycete fungus forms novel spiny structures and infects nematode.

    Science.gov (United States)

    Luo, Hong; Mo, Minghe; Huang, Xiaowei; Li, Xuan; Zhang, Keqin

    2004-01-01

    Nematophagous basidiomycete fungi kill nematodes by trapping, endoparasitizing and producing toxin. In our studies Coprinus comatus (O.F.Müll. : Fr.) Pers. is found to be a nematode-destroying fungus; this fungus immobilizes, kills and uses free-living nematode Panagrellus redivivus Goodey and root-knot nematode Meloidogyne arenaria Neal. C. comatus produces an unusual structure designated spiny ball. Set on a sporophore-like branch, the spiny ball is a burr-like structure assembled with a large number of tiny tubes. Purified spiny balls exhibit moderate nematicidal activity. Experiments show that spiny balls are not chlamydospores because of the absence of nuclei in the structures and quick formation within 3 d in a young colony. Nematodes added to C. comatus cultures on potato-dextrose agar (PDA) and cornmeal agar (CMA) become inactive in hours. Infection of nematodes by the fungus occurs only after the nematodes are immobilized (feeble or dead), probably by a toxin. Electron micrographs illustrate that C. comatus infect P. redivivus by producing penetration pegs with which hyphae colonize nematode bodies. An infected nematode is digested and consumed within days and hyphae grow out of the nematode.

  12. Coprinus comatus Damages Nematode Cuticles Mechanically with Spiny Balls and Produces Potent Toxins To Immobilize Nematodes▿

    Science.gov (United States)

    Luo, Hong; Liu, Yajun; Fang, Lin; Li, Xuan; Tang, Ninghua; Zhang, Keqin

    2007-01-01

    We reported recently a unique fungal structure, called the spiny ball, on the vegetative hyphae of Coprinus comatus (O. F. Müll.:Fr.) Pers. Although some observations regarding the role of this structure were presented, its function remained largely unknown. In this study, we showed that purified (isolated and washed) spiny balls could immobilize and kill the free-living nematode Panagrellus redivivus Goodey highly efficiently. Scanning electron microscopy studies illustrated that the spiny structure damaged the nematode cuticle, suggesting the presence of a mechanical force during the process of nematode immobilization. Severe injuries on nematode cuticles caused the leakage of inner materials of the nematodes. When these structures were ground in liquid nitrogen, their killing efficacy against nematodes was lost, indicating that the shape and the complete structure of the spiny balls are indispensable for their function. However, extraction with organic solvents never lowered their activity against P. redivivus, and the extracts showed no obvious effect on the nematode. We also investigated whether C. comatus was able to produce toxins which would aid in the immobilization of nematodes. In total, we identified seven toxins from C. comatus that showed activity to immobilize the nematodes P. redivivus and Meloidogyne incognita (Kofoid et White) Chitwood. The chemical structures of these toxins were identified with nuclear magnetic resonance, mass spectrometry, infrared, and UV spectrum analysis. Two compounds were found to be novel. The toxins found in C. comatus are O-containing heterocyclic compounds. PMID:17449690

  13. Perfil electroforético de las proteínas de la seta comestible "Coprinus comatus"

    OpenAIRE

    González Rodríguez, Andrea

    2017-01-01

    Este trabajo consiste en un estudio experimental del perfil proteico del hongo comestible Coprinus comatus, muy relevante por su alto valor económico y gastronómico, debido a sus propiedades nutricionales y medicinales. Los hongos son considerados alimentos funcionales, pues además de sus propiedades nutricionales, se han demostrado sus efectos beneficiosos para la salud que pueden ser utilizados en la prevención o tratamiento de enfermedades. Su acción terapéutica es atribuida a los compuest...

  14. Pluteus aurantiorugosus (Trog Sacc. and Coprinus patouillardii Quél. – new records of macrofungi for Romania

    Directory of Open Access Journals (Sweden)

    Mihai-Iulian Radu

    2013-12-01

    Full Text Available The paper gives a new contribution to the knowledge of macrofungi from Romania. Pluteus aurantiorugosus (Trog Sacc. is considered a rare macrofungus throughout Europe and this is the first report of this species in Romania. Coprinus patouillardii Quél. is a coprinoid species which is rarely mentioned, probably due to its small size, being usually overlooked. The species was uncovered and identified during a larger research regarding the diversity and distribution of macrofungi in the areas near Bucharest, Romania.

  15. A large pheromone and receptor gene complex determines multiple B mating type specificities in Coprinus cinereus.

    Science.gov (United States)

    O'Shea, S F; Chaure, P T; Halsall, J R; Olesnicky, N S; Leibbrandt, A; Connerton, I F; Casselton, L A

    1998-01-01

    Pheromone signaling plays an essential role in the mating and sexual development of mushroom fungi. Multiallelic genes encoding the peptide pheromones and their cognate 7-transmembrane helix (7-TM) receptors are sequestered in the B mating type locus. Here we describe the isolation of the B6 mating type locus of Coprinus cinereus. DNA sequencing and transformation analysis identified nine genes encoding three 7-TM receptors and six peptide pheromone precursors embedded within 17 kb of mating type-specific sequence. The arrangement of the nine genes suggests that there may be three functionally independent subfamilies of genes each comprising two pheromone genes and one receptor gene. None of the nine B6 genes showed detectable homology to corresponding B gene sequences in the genomic DNA from a B3 strain, and each of the B6 genes independently alter B mating specificity when introduced into a B3 host strain. However, only genes in two of the B6 groups were able to activate B-regulated development in a B42 host. Southern blot analysis showed that these genes failed to cross-hybridize to corresponding genes in the B42 host, whereas the three genes of the third subfamily, which could not activate development in the B42 host, did cross-hybridize. We conclude that cross-hybridization identifies the same alleles of a particular subfamily of genes in different B loci and that B6 and B42 share alleles of one subfamily. There are an estimated 79 B mating specificities: we suggest that it is the different allele combinations of gene subfamilies that generate these large numbers. PMID:9539426

  16. Heterologous expression and characterization of a novel laccase isoenzyme with dyes decolorization potential from Coprinus comatus.

    Science.gov (United States)

    Bao, Songyuan; Teng, Zhen; Ding, Shaojun

    2013-02-01

    Two new laccase genes, named lac1 and lac2, were cloned from the edible basidiomycete Coprinus comatus. Comparison of the deduced amino acid sequences revealed two laccases showed 66.12 % identity and clustered with lac2 and lac3 from Coprinopsis cinerea in same phylogenetic group. Lac1 and lac2 encode proteins of 517 and 523 amino acids preceded by 18 and 21-residue signal peptides, respectively. Lac1 was functionally expressed in Pichia pastoris. The optimum pHs of recombinant Lac1 were 3.0, 6.0, 5.5 and 6.0 and the optimum temperatures were 65, 55, 70 and 50 °C for ABTS, guaiacol, 2,6-dimethylphenol and syringaldazine, respectively. The Km values of Lac1 were 34, 4,317, 7,611 and 14 μM, and the corresponding kcat values were 465.79, 7.67, 1.15 and 0.60 (s(-1) mM), for ABTS, guaiacol, 2,6-dimethylphenol and syringaldazine, respectively. The enzyme activity was completely inhibited by sodium azide (NaN(3)) and 1,4-dithiothreitol (DTT) at the concentration of 5 mM. Laccase activity was also inhibited by several metal ions, especially Fe(2+), while K(+) and NH(4) (+) slightly enhanced laccase activity. Twelve synthetic dyes belonging to anthraquinone, azo and triphenylmethane dyes were decolorized by the recombinant Lac1 at different extents. The recombinant Lac1 decolorized azo dye Reactive Dark Blue KR up to 90 % without any mediator and increasing to 96 % with mediator, indicating its potential in the treatment of industrial effluent containing some recalcitrant synthetic dyes.

  17. Cytotoxic protein from the mushroom Coprinus comatus possesses a unique mode for glycan binding and specificity.

    Science.gov (United States)

    Zhang, Peilan; Li, Kunhua; Yang, Guang; Xia, Changqing; Polston, Jane E; Li, Gengnan; Li, Shiwu; Lin, Zhao; Yang, Li-Jun; Bruner, Steven D; Ding, Yousong

    2017-08-22

    Glycans possess significant chemical diversity; glycan binding proteins (GBPs) recognize specific glycans to translate their structures to functions in various physiological and pathological processes. Therefore, the discovery and characterization of novel GBPs and characterization of glycan-GBP interactions are significant to provide potential targets for therapeutic intervention of many diseases. Here, we report the biochemical, functional, and structural characterization of a 130-amino-acid protein, Y3, from the mushroom Coprinus comatus Biochemical studies of recombinant Y3 from a yeast expression system demonstrated the protein is a unique GBP. Additionally, we show that Y3 exhibits selective and potent cytotoxicity toward human T-cell leukemia Jurkat cells compared with a panel of cancer cell lines via inducing caspase-dependent apoptosis. Screening of a glycan array demonstrated GalNAcβ1-4(Fucα1-3)GlcNAc (LDNF) as a specific Y3-binding ligand. To provide a structural basis for function, the crystal structure was solved to a resolution of 1.2 Å, revealing a single-domain αβα-sandwich motif. Two monomers were dimerized to form a large 10-stranded, antiparallel β-sheet flanked by α-helices on each side, representing a unique oligomerization mode among GBPs. A large glycan binding pocket extends into the dimeric interface, and docking of LDNF identified key residues for glycan interactions. Disruption of residues predicted to be involved in LDNF/Y3 interactions resulted in the significant loss of binding to Jurkat T-cells and severely impaired their cytotoxicity. Collectively, these results demonstrate Y3 to be a GBP with selective cytotoxicity toward human T-cell leukemia cells and indicate its potential use in cancer diagnosis and treatment.

  18. The development of a thermostable CiP (Coprinus cinereus peroxidase) through in silico design.

    Science.gov (United States)

    Kim, Su Jin; Lee, Jeong Ah; Joo, Jeong Chan; Yoo, Young Je; Kim, Yong Hwan; Song, Bong Keun

    2010-01-01

    Protein thermostability is a crucial issue in the practical application of enzymes, such as inorganic synthesis and enzymatic polymerization of phenol derivatives. Much attention has been focused on the enhancement and numerous successes have been achieved through protein engineering methods. Despite fruitful results based on random mutagenesis, it was still necessary to develop a novel strategy that can reduce the time and effort involved in this process. In this study, a rapid and effective strategy is described for increasing the thermal stability of a protein. Instead of random mutagenesis, a rational strategy was adopted to theoretically stabilize the thermo labile residues of a protein using computational methods. Protein residues with high flexibility can be thermo labile due to their large range of movement. Here, residue B factor values were used to identify putatively thermo labile residues and the RosettaDesign program was applied to search for stable sequences. Coprinus cinereus (CiP) heme peroxidase was selected as a model protein for its importance in commercial applications, such as the polymerization of phenolic compounds. Eleven CiP residues with the highest B factor values were chosen as target mutation sites for thermostabilization, and then redesigned using RosettaDesign to identify sequences. Eight mutants based on the redesigns, were produced as functional enzymes and two of these (S323Y and E328D) showed increased thermal stability over the wild-type in addition to conserved catalytic activity. Thus, this strategy can be used as a rapid and effective in silico design tool for obtaining thermostable proteins. (c) 2010 American Institute of Chemical Engineers

  19. Transformation of multi-component ginkgolide into ginkgolide B by Coprinus comatus.

    Science.gov (United States)

    Ding, HongXue; Zhang, ZhiCai; Cao, ShengNan; Xu, Yin; Yu, JianGuo

    2015-03-14

    As the strongest antagonist of the platelet activating factor, ginkgolide B (GB) possesses anti-ischemic, anti-oxidant and anti-convulsant properties, and it is used for the treatment of thrombosis in clinical practice. Till now, GB is usually obtained from extraction of Ginkgo biloba leaves through column chromatography with an extremely low yield and high cost, which can not meet clinical requirement. Therefore, it is urgent to find a new method to prepare GB. In the current study, we studied the ability and mechanism to transform multi-component ginkgolide into GB by Coprinus comatus in order to enhance the GB yield. Except for ginkgolide A (GA) and GB, all the other ginkgolides in the extract were transformed by the strain. In the case of culture medium containing 20 g/L glucose, the transformation product was identified as 12% GA and 88% GB by high performance liquid chromatography-Mass spectrometry (HPLC-MS), two stage mass spectrometry (MS/MS) and nuclear magnetic resonance (NMR). Partial GA was also transformed into GB according to the yield (76%) and the content of GA in the raw ginkgolide (28.5%). Glucose was the key factor to transform ginkgolides. When glucose concentration in medium was higher than 40 g/L, all ginkgolides were transformed into the GB. Proteomic analysis showed that C. comatus transformed ginkgolide into GB by producing 5 aldo/keto reductases and catalases, and enhancing the metabolism of glucose, including Embden-Meyerhof pathway (EMP), hexose monophophate pathway (HMP) and tricarboxylic acid (TCA). C. comatus could transform ginkgolides into GB when the medium contained 40 g/L glucose. When the strain transformed ginkgolides, the glucose metabolism was enhanced and the strain synthesized more aldo/keto reductases and catalases. Our current study laid the groundwork for industrial production of GB.

  20. DNA ligase IV from a basidiomycete, Coprinus cinereus, and its expression during meiosis.

    Science.gov (United States)

    Namekawa, Satoshi; Ichijima, Yosuke; Hamada, Fumika; Kasai, Nobuyuki; Iwabata, Kazuki; Nara, Takayuki; Teraoka, Hirobumi; Sugawara, Fumio; Sakaguchi, Kengo

    2003-08-01

    DNA ligase IV is thought to be involved in DNA double-strand break repair and DNA non-homologous end-joining pathways, but these mechanisms are still unclear. To investigate the roles of DNA ligase IV from a biologically functional viewpoint, the authors studied its relationship to meiosis in a basidiomycete, Coprinus cinereus, which shows a highly synchronous meiotic cell cycle. The C. cinereus cDNA homologue of DNA ligase IV (CcLIG4) was successfully cloned. The 3.2 kb clone including the ORF encoded a predicted product of 1025 amino acid residues with a molecular mass of 117 kDa. A specific inserted sequence composed of 95 amino acids rich in aspartic acid and glutamic acid could be detected between tandem BRCT domains. The inserted sequence had no sequence identity with other eukaryotic counterparts of DNA ligase IV or with another aspartic acid and glutamic acid rich sequence inserted in C. cinereus proliferating cell nuclear antigen (CcPCNA), although the length and the percentages of aspartic and glutamic acids were similar. In addition, the recombinant CcLIG4 protein not only showed ATP-dependent ligase activity, but also used (dT)(16)/poly(dA) and (dT)(16)/poly(rA) as substrates, and had double-strand ligation activity, like human DNA ligase IV. Northern hybridization analysis and in situ hybridization indicated that CcLIG4 was expressed not only at the pre-meiotic S phase but also at meiotic prophase I. Intense signals were observed in leptotene and zygotene. Based on these observations, the possible role(s) of C. cinereus DNA ligase IV during meiosis are discussed.

  1. Consumption of Coprinus comatus polysaccharide extract causes recovery of alcoholic liver damage in rats.

    Science.gov (United States)

    Ozalp, Fatma Ozgul; Canbek, Mediha; Yamac, Mustafa; Kanbak, Gungor; Van Griensven, Leo J L D; Uyanoglu, Mustafa; Senturk, Hakan; Kartkaya, Kazım; Oglakci, Aysegul

    2014-08-01

    Excess use of alcohol is known to be associated with liver diseases such as fatty liver, alcoholic hepatitis, and cirrhosis. Various practices may be applied to prevent or treat the damage caused by chronic alcoholism. Coprinus comatus (O.F. Müll.) Pers. (Agaricaceae) is a macrofungus that has been reported to aid the recovery of murine livers damaged by benzopyrene. In this study, the possible therapeutic effects of three different doses (50, 100, and 150 mg/kg) of C. comatus polysaccharide (PS) extract were studied in rats subjected to an alcoholic diet. The histological and biochemical results were compared between the control and experimental groups. Modified Lieber-Decarli's calorie-adjusted liquid alcohol diet was given orally for 60 d. In addition to histopathology, alanine transaminase (ALT), aspartate transaminase (AST), mitochondrial membrane integrity, total cytochrome-c oxidase activity (TotalStCox), total mitochondrial cytochrome-c oxidase activity (TotalMtStCox), and caspase-3 values were used as liver parameters, and liver sections from all experimental groups were examined by electron microscopy. Using histopathological assessment, it was observed that there was a decline in liver hepatocyte vacuolization in the treatment group fed 50 mg PS/kg. The TotalStCox and TotalMtStCox values of this group differed from the EtOH control group (p < 0.05). Daily administration of 50 mg/kg of C. comatus PS extract considerably reduced the negative effects of alcohol on liver structure and function.

  2. Google Secrets

    CERN Document Server

    Davis, Yvette

    2011-01-01

    Become a Google guru with these effective tips, tricks, and techniques Sure, you use Google. But do you really use Google-and everything it has to offer-in the most effective way possible? Wish you could just sit down with a Google expert who would show you how to take your Google savviness to the next level? With Google Secrets, you can! Tech expert Jerri Ledford reveals the ins, outs, and little-known facts about Google to show you how to sharpen your skills so you can get more done, more efficiently. You may already be familiar with Google's most popular applications, but this indispensable

  3. Coprinus comatus cap inhibits adipocyte differentiation via regulation of PPARγ and Akt signaling pathway.

    Science.gov (United States)

    Park, Hyoung Joon; Yun, Jisoo; Jang, Sun-Hee; Kang, Suk Nam; Jeon, Beong-Sam; Ko, Yeoung-Gyu; Kim, Hong-Duck; Won, Chung-Kil; Kim, Gon-Sup; Cho, Jae-Hyeon

    2014-01-01

    This study assessed the effects of Coprinus comatus cap (CCC) on adipogenesis in 3T3-L1 adipocytes and the effects of CCC on the development of diet-induced obesity in rats. Here, we showed that the CCC has an inhibitory effect on the adipocyte differentiation of 3T3-L1 cells, resulting in a significant decrease in lipid accumulation through the downregulation of several adipocyte specific-transcription factors, including CCAAT/enhancer binding protein β, C/EBPδ, and peroxisome proliferator-activated receptor gamma (PPARγ). Moreover, treatment with CCC during adipocyte differentiation induced a significant down-regulation of PPARγ and adipogenic target genes, including adipocyte protein 2, lipoprotein lipase, and adiponectin. Interestingly, the CCC treatment of the 3T3-L1 adipocytes suppressed the insulin-stimulated Akt and GSK3β phosphorylation, and these effects were stronger in the presence of an inhibitor of Akt phosphorylation, LY294002, suggesting that CCC inhibited adipocyte differentiation through the down-regulation of Akt signaling. In the animal study, CCC administration significantly reduced the body weight and adipose tissue weight of rats fed a high fat diet (HFD) and attenuated lipid accumulation in the adipose tissues of the HFD-induced obese rats. The size of the adipocyte in the epididymal fat of the CCC fed rats was significantly smaller than in the HFD rats. CCC treatment significantly reduced the total cholesterol and triglyceride levels in the serum of HFD rats. These results strongly indicated that the CCC-mediated decrease in body weight was due to a reduction in adipose tissue mass. The expression level of PPARγ and phospho-Akt was significantly lower in the CCC-treated HFD rats than that in the HFD obesity rats. These results suggested that CCC inhibited adipocyte differentiation by the down-regulation of major transcription factor involved in the adipogenesis pathway including PPARγ through the regulation of the Akt pathway in 3T3

  4. Coprinus comatus cap inhibits adipocyte differentiation via regulation of PPARγ and Akt signaling pathway.

    Directory of Open Access Journals (Sweden)

    Hyoung Joon Park

    Full Text Available This study assessed the effects of Coprinus comatus cap (CCC on adipogenesis in 3T3-L1 adipocytes and the effects of CCC on the development of diet-induced obesity in rats. Here, we showed that the CCC has an inhibitory effect on the adipocyte differentiation of 3T3-L1 cells, resulting in a significant decrease in lipid accumulation through the downregulation of several adipocyte specific-transcription factors, including CCAAT/enhancer binding protein β, C/EBPδ, and peroxisome proliferator-activated receptor gamma (PPARγ. Moreover, treatment with CCC during adipocyte differentiation induced a significant down-regulation of PPARγ and adipogenic target genes, including adipocyte protein 2, lipoprotein lipase, and adiponectin. Interestingly, the CCC treatment of the 3T3-L1 adipocytes suppressed the insulin-stimulated Akt and GSK3β phosphorylation, and these effects were stronger in the presence of an inhibitor of Akt phosphorylation, LY294002, suggesting that CCC inhibited adipocyte differentiation through the down-regulation of Akt signaling. In the animal study, CCC administration significantly reduced the body weight and adipose tissue weight of rats fed a high fat diet (HFD and attenuated lipid accumulation in the adipose tissues of the HFD-induced obese rats. The size of the adipocyte in the epididymal fat of the CCC fed rats was significantly smaller than in the HFD rats. CCC treatment significantly reduced the total cholesterol and triglyceride levels in the serum of HFD rats. These results strongly indicated that the CCC-mediated decrease in body weight was due to a reduction in adipose tissue mass. The expression level of PPARγ and phospho-Akt was significantly lower in the CCC-treated HFD rats than that in the HFD obesity rats. These results suggested that CCC inhibited adipocyte differentiation by the down-regulation of major transcription factor involved in the adipogenesis pathway including PPARγ through the regulation of the

  5. Heptemerones A-G, seven novel diterpenoids from Coprinus heptemerus: producing organism, fermentation, isolation and biological activities.

    Science.gov (United States)

    Kettering, Melanie; Valdivia, Carola; Sterner, Olov; Anke, Heidrun; Thines, Eckhard

    2005-06-01

    Seven novel diterpenoids, named heptemerones A-G, were isolated from the broth of submerged cultures of Coprinus heptemerus, a basidiomycete which previously had not been known to produce secondary metabolites. The compounds were purified by solid phase extraction and silica gel chromatography followed by preparative HPLC. Among the biological activities the inhibition of fungal germination was the most potent, and depended highly on the composition of the assay medium. In water, inhibition occurred at 5 - 10 fold lower concentrations as compared to complex media. Heptemerone G was the most active compound with MICs starting at 1 microg/ml. Four of the antifungal compounds exhibited plant protective activity in a leaf segment assay using Magnaporthe grisea as the pathogen. Growth of yeasts and bacteria was hardly affected. Cytotoxic activities were moderate and only heptemerone D was phytotoxic.

  6. Primary structure of dihydrofolate reductase and mitochondrial ribosomal protein L36 genes from the basidiomycete Coprinus cinereus.

    Science.gov (United States)

    Aimi, Tadanori; Fukuhara, Shoji; Ishiguro, Maki; Kitamoto, Yutaka; Morinaga, Tsutomu

    2004-08-01

    We amplified and sequenced the dihydrofolate reductase (DHFR) gene of the basidiomycete Coprinus cinereus. Downstream of the DHFR coding region, a mitochondrial (mt) ribosomal protein L36 (RPL36) gene was discovered in the opposite orientation to DHFR gene. Putative polyadenylation signals of the two genes overlapped, both containing the 8-bp palindrome 5'-aatatatt-3'. The finding that C. cinereus DHFR gene is closely clustered with a mt protein gene strongly suggests that C. cinereus DHFR is closely related to mt function and evolution. The amino acid sequence of C. cinereus DHFR is most homologous to eukaryotic proteins such as Cryptococcus neoformans and Pneumocystis carinii DHFRs. However, the sequence of C. cinereus mt RPL36 closely resembles RPL36 of bacteria and cyanobacteria such as Synechocystis sp. and Escherichia coli. This result strongly supports the serial endosymbiotic theory of the development of ancestral eukaryotes, and suggests that C. cinereus mt RPL36 gene originated from the ancestral eubacterial genome.

  7. A constitutively active G-protein-coupled receptor causes mating self-compatibility in the mushroom Coprinus.

    Science.gov (United States)

    Olesnicky, N S; Brown, A J; Dowell, S J; Casselton, L A

    1999-01-01

    In the mushroom Coprinus cinereus, the multiallelic B mating type genes are predicted to encode a large family of seven-transmembrane domain receptors and CaaX-modified pheromones. We have shown that a single amino acid change Q229P in transmembrane domain VI of one receptor confers a self-compatible mating phenotype. Using a heterologous yeast assay, we have demonstrated that this C.cinereus pheromone receptor is a G-protein-coupled receptor and that the Q229P mutation is constitutively activating. A C.cinereus pheromone precursor was processed to an active species specifically in yeast MATa cells and activated the co-expressed wild-type receptor. Yeast cells expressing the wild-type receptor were used to test the activity of synthetic peptides, enabling us to predict the structure of the mature C.cinereus pheromone and to show that the Q229P mutation does not compromise normal receptor function. PMID:10329622

  8. Expression of multi-functional cellulase gene mfc in Coprinus cinereus under control of different basidiomycete promoters.

    Science.gov (United States)

    Cheng, Shujie; Yang, Peizhou; Guo, Liqiong; Lin, Junfang; Lou, Nannan

    2009-10-01

    Multi-functional cellulase gene mfc was expressed in Coprinus cinereus under naturally non-inductive conditions using three heterologous promoters. Endo-beta-1,4-glucanase expression was achieved in solid and liquid media with promoter sequences from the Lentinula edodesgpd gene, the Flammulina velutipes gpd gene and the Volvariella volvaceagpd gene. As measured by enzyme activity in liquid cultures, a 613-bp gpd promoter fragment from L. edodes was most efficient, followed by a 752-bp gpd fragment from F. velutipes. The V. volvacea gpd promoter sequence was less active, in comparison. Irrespective of the promoter used, enzymatic activities increase 34-fold for highly active transformants and 29-fold for less active one by using cellulase-inducing medium. The highest activities of endo-beta-1,4-glucanase (34.234 U/ml) and endo-beta-1,4-xylanase (263.695 U/ml) were reached by using the L. edodesgpd promoter.

  9. Comparison of hypoglycemic activity and toxicity of vanadium (IV) and vanadium (V) absorbed in fermented mushroom of Coprinus comatus.

    Science.gov (United States)

    Ma, Zhaoji; Fu, Qin

    2009-12-01

    This study was designed to evaluate the effect and toxicity of administration of vanadium (IV, V) absorbed by Coprinus comatus (VACC) on alloxan-induced and sucrosefed hyperglycemic mice, respectively. The blood glucose, lipid profile, and the organ masses of the mice were analyzed. After the mice were administered with VACC, the blood glucose and the lipid profile of hyperglycemic mice decreased, irrespective of the VACC produced by vanadium (IV) or vanadium (V). However, the organ masses of the mice were significantly different after the mice were treated with vanadium (IV) and vanadium (V) 9 weeks later. The results indicate both vanadium (IV) and vanadium (V) absorbed in C. comatus have hypoglycemic activity on hyperglycemic mice. However, vanadium (IV) absorbed in C. comatus is less toxic to mice than vanadium (V).

  10. The polysaccharide extracts from the fungi Coprinus comatus and Coprinellus truncorum do exhibit AChE inhibitory activity.

    Science.gov (United States)

    Pejin, Boris; Tešanović, Kristina; Jakovljević, Dragica; Kaišarević, Sonja; Šibul, Filip; Rašeta, Milena; Karaman, Maja

    2017-11-22

    The polysaccharide (PSH) extracts from the edible mushroom species Coprinus comatus and Coprinellus truncorum were screened in liquid for their acetylcholinesterase inhibitory (AChE) activity. Both extracts were found to display inhibition of the aforementioned enzyme reaching similar IC50 values of 0.62 ± 0.07 and 0.61 ± 0.03 mg/mL, respectively. According to the means of FTIR spectroscopy, these PSH extracts mostly contained β-glucans. However, the presence of some proteins and polyphenolics as minor ingredients were also detected. Compared with existing literature data for anti-AChE activity of the sugar samples, the findings within this study may be treated as a profound bioactivity. Consequently, this study puts some light on the possible use of the screened macrofungi in the palliative treatment of Alzheimer's disease.

  11. Production of Oxidative and Hydrolytic Enzymes by Coprinus cinereus (Schaeff.) Gray from Sisal Wastes Supplemented with Cow Dung Manure

    Science.gov (United States)

    Raymond, Prosper; Mshandete, Anthony Manoni; Kajumulo Kivaisi, Amelia

    2015-01-01

    The activity of oxidative and hydrolytic enzymes of the edible and medicinal white rot fungi Coprinus cinereus (Schaeff.) Gray mushroom was observed during mycelia growth and fruiting body development in solid substrate fermentation using sisal waste fractions amended with cow dung manure as supplement. Laccase had the highest titre value among the five detected enzymes. Its activity was higher during mycelia growth compared to fruiting phase, with 10% supplemented substrate formulation unmixed sisal leaf decortication residues [abbreviated SL : SB (100 : 0)] displaying the highest activity of 39.45 ± 12.05 Ug−1. Lignin peroxidase (LiP) exhibited a characteristic wave-like pattern with the highest peaks found either during full mycelia colonization or soon after first flush harvest; the highest activity of 1.93 ± 0.62 Ug−1 was observed on unsupplemented SL : SB (100 : 0) substrate formulation during mycelia colonization. For hydrolytic enzymes, the highest carboxymethyl cellulase (CMCase) activity of 2.03 ± 0.70 Ug−1 was observed on 20% supplemented SL : SB (0 : 100) after first flush; that of pectinase (1.90 ± 0.32 Ug−1) was revealed after third flush on 10% supplemented SL : SB (0 : 100) substrate formulation while 10% supplemented SL : SB (25 : 75) exhibited the highest xylanase activity (1.23 ± 0.12 Ug−1) after first flush. These findings show that the activities of both oxidative and hydrolytic enzymes were regulated in line with developmental phase of growth of Coprinus cinereus. PMID:26664748

  12. Production of Oxidative and Hydrolytic Enzymes by Coprinus cinereus (Schaeff. Gray from Sisal Wastes Supplemented with Cow Dung Manure

    Directory of Open Access Journals (Sweden)

    Prosper Raymond

    2015-01-01

    Full Text Available The activity of oxidative and hydrolytic enzymes of the edible and medicinal white rot fungi Coprinus cinereus (Schaeff. Gray mushroom was observed during mycelia growth and fruiting body development in solid substrate fermentation using sisal waste fractions amended with cow dung manure as supplement. Laccase had the highest titre value among the five detected enzymes. Its activity was higher during mycelia growth compared to fruiting phase, with 10% supplemented substrate formulation unmixed sisal leaf decortication residues [abbreviated SL : SB (100 : 0] displaying the highest activity of 39.45±12.05 Ug−1. Lignin peroxidase (LiP exhibited a characteristic wave-like pattern with the highest peaks found either during full mycelia colonization or soon after first flush harvest; the highest activity of 1.93±0.62 Ug−1 was observed on unsupplemented SL : SB (100 : 0 substrate formulation during mycelia colonization. For hydrolytic enzymes, the highest carboxymethyl cellulase (CMCase activity of 2.03±0.70 Ug−1 was observed on 20% supplemented SL : SB (0 : 100 after first flush; that of pectinase (1.90±0.32 Ug−1 was revealed after third flush on 10% supplemented SL : SB (0 : 100 substrate formulation while 10% supplemented SL : SB (25 : 75 exhibited the highest xylanase activity (1.23±0.12 Ug−1 after first flush. These findings show that the activities of both oxidative and hydrolytic enzymes were regulated in line with developmental phase of growth of Coprinus cinereus.

  13. Screening of Coprinus species for the production of extracellular peroxidase and evaluation of its applicability to the treatment of aqueous phenol

    Energy Technology Data Exchange (ETDEWEB)

    Ikehata, K.; Buchanan, I.D. [Univ. of Alberta, Dept. of Civil and Environmental Engineering, Edmonton, Alberta (Canada)

    2002-06-15

    Twenty-nine strains of Coprinus species comprising 16 strains from 12 identified species and 13 unidentified strains as well as one Arthromyces ramosus strain were screened for the production of extracellular peroxidase. Among the fungi examined, three strains of C. cinereus, UAMH 4103, UAMH 7907 and IFO 30116, as well as one Coprinus sp., UAMH 10067, which was isolated from urea treated soil, were shown to produce large amounts of extracellular peroxidase. The performance of crude peroxidase, obtained from liquid culture of C. cinereus, (CIP) on phenol removal from synthetic wastewater was evaluated and compared with that of purified horseradish peroxidase and A. ramosus peroxidase. Although crude CIP performed better than both purified enzymes, its superiority vanished in the presence of poly(ethylene glycol), a known protective agent of peroxidase. This suggests that the residual soluble substances present in crude CIP have protective effects similar to those of poly(ethylene glycol). (author)

  14. [Cloning of y3 gene encoding a tobacco mosaic virus inhibitor from Coprinus comatus and transformation to Nicotiana tabacum].

    Science.gov (United States)

    Wang, Xueren; He, Tao; Zhang, Gaina; Hao, Jianguo; Jia, Jingfen

    2010-02-01

    The protein Y3 was a TMV inhibitor which was encoded by y3 gene. The aim of this work was to clone the full length of y3 gene from Coprinus comatus and to reveal its inhibitory function to TMV in in vivo conditions. We amplified the unknown 5'- terminal cDNA sequence of y3 gene with 5'- Full RACE Core Set (TaKaRa), obtained the full length of this gene by RT-PCR, constructed the expression plasmid pCAMBIA1301-y3 via inserting gene y3 sequence, CaMV 35 S promoter, and NOS terminator at MCS and transformed it into Nicotiana tabacum via agrobacterium-mediation. The full length of y3 gene was 534 bps including one ORF encoding 130 amino acid residues (GenBank Accession No. GQ859168; EMBL FN546262). The cDNA sequence and its deduced amino acid sequence showed high similarity (94%) to the published fragment of y3 gene sequence. Northern blot analysis proved the transcription of y3 gene in transgenic tobacco plants. The transgenic plants inoculated with TMV expressed the inhibitory activity to TMV. We cloned the full length of y3 gene and obtained transgenic tobacco plants. The expression of y3 gene in transgenic plants improved the inhibitory activity to TMV. The cloning and expression analysis of y3 gene might provide background information for future studying of y3 gene.

  15. Knockdown of LIM15/DMC1 in the mushroom Coprinus cinereus by double-stranded RNA-mediated gene silencing.

    Science.gov (United States)

    Namekawa, Satoshi H; Iwabata, Kazuki; Sugawara, Hiroko; Hamada, Fumika N; Koshiyama, Akiyo; Chiku, Hiroyuki; Kamada, Takashi; Sakaguchi, Kengo

    2005-11-01

    The basidiomycete Coprinus cinereus has many advantages as a model organism for studying sexual development and meiosis, but it has been difficult to investigate using reverse-genetics methods, such as gene disruption by homologous recombination. Here, gene repression by dsRNA-mediated gene silencing was tried as an alternative method for reverse-genetics studies. It was shown that transformation of the LIM15/DMC1 dsRNA expression construct (LIM15dsRNA) resulted in genomic insertion of LIM15dsRNA and paucity of the LIM15/DMC1 transcript. First, LIM15dsRNA was transformed into the homothallic strain AmutBmut to generate a homozygote in which both nuclei had a copy of LIM15dsRNA. The LIM15/DMC1-repressed strain showed abnormal homologous chromosome synapsis during meiosis. Basidiospore production was reduced to 16 % by the induction of dsRNA. However, approximately 60 % of basidiospores were viable. Next, a heterozygote was generated in which one nucleus had a copy of LIM15dsRNA. The phenotype was similar to that of the homozygote. These results are not only the first demonstration of dsRNA-mediated gene silencing in a member of the homobasidiomycete fungi, to which 90 % of mushroom species belong, but also the first successful use of a reverse-genetics approach in C. cinereus research.

  16. A Novel Laccase with Potent Antiproliferative and HIV-1 Reverse Transcriptase Inhibitory Activities from Mycelia of Mushroom Coprinus comatus

    Directory of Open Access Journals (Sweden)

    Shuang Zhao

    2014-01-01

    Full Text Available A novel laccase was isolated and purified from fermentation mycelia of mushroom Coprinus comatus with an isolation procedure including three ion-exchange chromatography steps on DEAE-cellulose, CM-cellulose, and Q-Sepharose and one gel-filtration step by fast protein liquid chromatography on Superdex 75. The purified enzyme was a monomeric protein with a molecular weight of 64 kDa. It possessed a unique N-terminal amino acid sequence of AIGPVADLKV, which has considerably high sequence similarity with that of other fungal laccases, but is different from that of C. comatus laccases reported. The enzyme manifested an optimal pH value of 2.0 and an optimal temperature of 60°C using 2,2′-azinobis(3-ethylbenzothiazolone-6-sulfonic acid diammonium salt (ABTS as the substrate. The laccase displayed, at pH 2.0 and 37°C, Km values of 1.59 mM towards ABTS. It potently suppressed proliferation of tumor cell lines HepG2 and MCF7, and inhibited human immunodeficiency virus type 1 (HIV-1 reverse transcriptase (RT with an IC50 value of 3.46 μM, 4.95 μM, and 5.85 μM, respectively, signifying that it is an antipathogenic protein.

  17. Systemic treatment with vanadium absorbed by Coprinus comatus promotes femoral fracture healing in streptozotocin-diabetic rats.

    Science.gov (United States)

    Wang, Guangbin; Wang, Jiashi; Fu, Yonghui; Bai, Lunhao; He, Ming; Li, Bin; Fu, Qin

    2013-03-01

    The purpose of this study was to analyze the impact of vanadium absorbed by Coprinus comatus (VACC) on fracture healing in streptozotocin-diabetic rats. Forty-five male Wistar rats used were divided into three groups: normal rats (control), diabetic rats, and diabetic rats treated with VACC. A standardized fracture-healing model with a stable plate fixation was established for the rat femoral fracture. After a 4-week stable fixation, callus quality was assessed by microcomputerized tomography and histological and biomechanical examinations. In addition, bone samples were obtained to evaluate the content of mineral substances in bones. Compared with the diabetic group, vanadium treatment significantly increased bone mineral content and biomechanical strength and improved microstructural properties of the callus. The ultimate load was increased by 29.1 % (P<0.05), and the total bone volume of callus enhanced by 11.2 % (P<0.05) at 4 weeks post fracture. Vanadium also promoted callus bone formation, which caused a 35.5 % increase in the total area of callus. However, VACC did not accelerate the fracture repair process in histological analysis. In conclusion, the current study suggests that systemic treatment with vanadium could promote fracture healing in streptozotocin-diabetic rats.

  18. The rad9 gene of Coprinus cinereus encodes a proline-rich protein required for meiotic chromosome condensation and synapsis

    Energy Technology Data Exchange (ETDEWEB)

    Seitz, L.C.; Tang, Keliang; Cummings, W.J.; Zolan, M.E. [Indiana Univ., Bloomington, IN (United States)

    1996-04-01

    The rad9 gene of Coprinus cinereus is essential for the normal completion of meiosis. We examined surface-spread preparations of wild-type and rad9-1 nuclei from the meiotic stages of karyogamy through metaphase I, and we determined the primary sequence, structure, and meiotic expression of the rad9 gene. In wild-type C. cinereus, karyogamy is followed by condensation and alignment of homologous chromosomes. Condensation and axial core development largely precede synapsis, which often initiates at telomeres. A diffuse diplotene phase coincides with dissolution of the synaptonemal complex, and subsequently chromosomes further condense as the cells progress into metaphase I. In contrast, although karyogamy and nucleolar fusion are apparently normal in rad9-1 basidia, only short stretches of synaptonemal complex form. These correlate with stretches of condensed chromatin, mostly at apparent chromosome ends, and regions of presumptive triple synapsis are numerous. rad9-1 basidia enter the diffuse stages of early diplotene, and then 50% of these cells enter metaphase I by the criteria of nucleolar elimination and at least some chromatin condensation. rad9 gene expression is induced after gamma irradiation and during meiosis. The gene has 27 exons and encodes a predicted protein of 2157 amino acids, with a proline-rich amino terminus. 62 refs., 10 figs.

  19. Molecular analysis of pcc1, a gene that leads to A-regulated sexual morphogenesis in Coprinus cinereus.

    Science.gov (United States)

    Murata, Y; Fujii, M; Zolan, M E; Kamada, T

    1998-01-01

    A homokaryotic strain (5337) in our culture stock of Coprinus cinereus produced fertile fruit bodies after prolonged culture. Microscopic examination revealed that hyphae dedifferentiated from the tissues of one of the fruit bodies, as well as all basidiospore derivatives from the fruit body, exhibited pseudoclamps, whereas vegetative hyphae of 5337, from which the fruit body developed, had no clamp connections. Genetic analysis showed that the formation of pseudoclamps results from a recessive mutation in a gene designated pcc1 (pseudoclamp connection formation), which is distinct from the A and B mating type genes. Cloning and sequencing of the pcc1 gene and cDNA identified an ORF of 1683 bp interrupted by one intron. Database searches revealed that pcc1 encodes an SRY-type HMG protein. The HMG box shared 44, 41, and 29% sequence identities (>80 amino acids) to those of FPR1 of Podospora anserina, MAT-Mc of Schizosaccharomyces pombe, and prf1 of Ustilago maydis, respectively. Northern analysis revealed that the level of pcc1 expression is higher in the dikaryon, in homokaryons in which the A and B mating type developmental sequences are individually activated, than in the homokaryon in which these sequences are not active. Sequencing of the pcc1-1 mutant allele revealed that the mutant carries a nonsense mutation at serine 211, a residue located between the HMG box and the C terminus. Based on these results, possible roles of the pcc1 gene in the sexual development of homobasidiomycetes are discussed. PMID:9691034

  20. The dst1 gene involved in mushroom photomorphogenesis of Coprinus cinereus encodes a putative photoreceptor for blue light.

    Science.gov (United States)

    Terashima, Kazuhisa; Yuki, Katsuyuki; Muraguchi, Hajime; Akiyama, Masashi; Kamada, Takashi

    2005-09-01

    The homobasidiomycete Coprinus cinereus exhibits remarkable photomorphogenesis during fruiting-body development. Under proper light conditions, fruiting-body primordia proceed to the maturation phase in which basidia in the pileus undergo meiosis, producing sexual spores, followed by stipe elongation and pileus expansion for efficient dispersal of the spores. In the continuous darkness, however, the primordia do not proceed to the maturation phase but are etiolated: the pileus and stipe tissues at the upper part of the primordium remain rudimentary and the basal part of the primordium elongates, producing "dark stipe." In this study we genetically analyzed five strains that produce dark stipes even if light conditions promoting the maturation are given and then characterized one of them, Uar801 (dst1-1). The dst1 gene was cloned as a DNA fragment that rescues the dst1-1 mutation. Dst1 is predicted to be a protein of 1175 amino acids that contains two PAS domains, a coiled-coil structure, and a putative, glutamine-rich, transcriptional activation domain (AD). One of the PAS domains exhibits significant similarity to the LOV domains of known blue-light receptors, suggesting that Dst1 is a blue-light receptor of C. cinereus. The dst1-1 mutation is predicted to truncate the putative AD in the C-terminal region.

  1. Development of the radical-stable Coprinus cinereus peroxidase (CiP) by blocking the radical attack.

    Science.gov (United States)

    Kim, Su Jin; Joo, Jeong Chan; Kim, Han Sang; Kwon, Inchan; Song, Bong Keun; Yoo, Young Je; Kim, Yong Hwan

    2014-11-10

    Despite the potential use of peroxidases as industrial biocatalysts, their practical application is often impeded due to suicide inactivation by radicals generated in oxidative reactions. Using a peroxidase from Coprinus cinereus (CiP) as a model enzyme, we revealed a dominant factor for peroxidase inactivation during phenol oxidation, and we engineered radical-stable mutants by site-directed mutagenesis of an amino acid residue susceptible to modification by phenoxyl radical. Mass spectrometry analysis of inactivated CiP identified an adduct between F230 and a phenoxyl radical, and subsequently, the F230 residue was mutated to amino acids that resisted radical coupling. Of the F230 mutants, the F230A mutant showed the highest stability against radical inactivation, retaining 80% of its initial activity, while the wild-type protein was almost completely inactivated. The F230A mutant also exhibited a 16-fold higher turnover of the phenol substrate compared with the wild-type enzyme. Furthermore, the F230A mutant was stable during the oxidation of other phenolic compounds, including m-cresol and 3-methoxyphenol. No structural changes were observed by UV-vis and CD spectra of CiP after radical coupling, implying that the F230-phenol radical adduct inactivated CiP by blocking substrate access to the active site. Our novel strategy can be used to improve the stability of other peroxidases inactivated by radicals. Copyright © 2014 Elsevier B.V. All rights reserved.

  2. Strengthening detoxication impacts of Coprinus comatus on nickel and fluoranthene co-contaminated soil by bacterial inoculation.

    Science.gov (United States)

    Tang, Xia; Liu, Bingxin; Deng, Qiusui; Zhang, Ruitao; Li, Xue; Xu, Heng

    2017-11-10

    To develop an efficient and environmental-friendly approach to detoxicate nickel (Ni) and fluoranthene co-contaminated soil, the combined application of Coprinus comatus (C. comatus) with Serratia sp. FFC5 and/or Enterobacter sp. E2 was investigated. The pot experiment tested the influences of bacterial inoculation on the growth of C. comatus, content of Ni in C. comatus, Ni speciation in soil, fluoranthene dissipation, soil enzymatic activities, bacterial population and community structure. With the inoculation of bacteria, the fresh weights of C. comatus, concentration of Ni in C. comatus and the dissipation rates of fluoranthene were increased by 17.73-29.38%, 68.97-204.97% and 34.84-60.90%, respectively. Notably, results illustrated that the co-inoculation of FFC5 and E2 showed better effect in biomass enhancement, Ni accumulation and fluoranthene dissipation than solitary inoculation. Simultaneously, higher soil enzymatic and microbiological activities suggested that the integrated detoxication method of bacteria and C. comatus could improve soil quality. Therefore, we can infer that bacterial inoculation strengthened detoxication effect of C. comatus in Ni-fluoranthene co-contaminated soil, indicating that the combined application of C. comatus and bacteria can be an efficient alternative for detoxicating Ni and fluoranthene co-contaminated soil. Copyright © 2017. Published by Elsevier Ltd.

  3. DNA polymerase mu interacts with a meiosis-specific RecA homolog Lim15 during meiosis in Coprinus cinereus.

    Science.gov (United States)

    Yanai, Takuro; Sakamoto, Aiko; Iwabata, Kazuki; Koshiyama, Akiyo; Sugawara, Hiroko; Nara, Takayuki; Takakusagi, Yoichi; Sakaguchi, Kengo

    2009-12-04

    Meiosis is a fundamental process in eukaryotes. Homologous chromosomes are paired and recombined during meiotic prophase I, which results in variation among the gametes. However, the mechanism of recombination between the maternal and paternal chromosome is unknown. In this study, we report on the identification of interaction between Coprinus cinereus DNA polymerase mu (CcPol mu) and CcLim15/Dmc1, a meiosis-specific RecA-like protein, during meiosis. Interaction between these two proteins was confirmed using a GST-pull down assay. A two-hybrid assay revealed that the N-terminus of CcPol mu, which includes the BRCT domain, is responsible for binding the C-terminus of CcLim15. Furthermore, co-immunoprecipitation experiments indicate that these two proteins also interact in the crude extract of the meiotic cell. A significant proportion of CcPol mu and CcLim15 is shown to co-localize in nuclei from the leptotene/zygotene stage to the early pachytene stage during meiotic prophase I. Moreover, CcLim15 enhances polymerase activity of CcPol mu early in the reaction. These results suggest that CcPol mu might be recruited by CcLim15 and elongate the D-loop structure during homologous recombination in meiosis.

  4. The effects of vanadium (V) absorbed by Coprinus comatus on bone in streptozotocin-induced diabetic rats.

    Science.gov (United States)

    Pei, Yi; Fu, Qin

    2011-09-01

    The purpose of this study was to evaluate the effects of vanadium absorbed by Coprinus comatus (VACC) treatment on bone in streptozotocin (STZ)-induced diabetic rats. Forty-five Wistar female rats used were divided into three groups: (1) normal rats (control), (2) diabetic rats, and (3) diabetic rats treated with VACC. Normal and diabetic rats were given physiological saline, and VACC-treated rats were administered VACC intragastrically at doses of 0.18 mg vanadium/kg body weight once daily. Treatments were performed over a 12-week period. At sacrifice, one tibia and one femur were removed, subjected to micro computed tomography (micro-CT) for determination of trabecular bone structure, and then processed for histomorphometry to assess bone turnover. Another femoral was used for mechanical testing. In addition, bone samples were collected to evaluate the content of mineral substances in bones. Treatment with VACC increased trabecular bone volume fraction in diabetic rats. Vanadium-treated animals had significant increases in ultimate load, trabecular thickness, and osteoblast surface. However, vanadium treatment did not seem to affect bone stiffness, bone energy absorption, trabecular separation, and osteoclast number. P levels in the femurs of diabetic rats treated with VACC were significantly higher than those of diabetic animals. Ca levels in diabetic and diabetic rats treated with vanadium showed no obvious changes. In conclusion, our results provide an important proof of concept that VACC may represent a powerful approach to treating or reversing diabetic osteopathy in humans.

  5. Efficient isolation and mapping of rad genes of the fungus Coprinus cinereus using chromosome-specific libraries.

    Science.gov (United States)

    Zolan, M E; Crittenden, J R; Heyler, N K; Seitz, L C

    1992-01-01

    We have constructed cosmid libraries from electrophoretically separated chromosomes of the basidiomycete Coprinus cinereus. These libraries greatly facilitate the isolation of genes by complementation of mutant phenotypes and are particularly useful for map-based cloning strategies. From a library constructed from two co-migrating C.cinereus chromosomes, we isolated a clone that complements the C.cinereus rad9-1 mutation. Examination of this clone showed that it complements both the repair and meiotic defects of this mutant. Restriction fragment length polymorphism mapping using a portion of this clone showed that it maps to the rad9 locus. In addition, a single copy of transforming DNA is sufficient to complement the rad9-1 defects. Thus, we believe we have cloned the rad9 gene itself. We also used a chromosome-specific library and backcrossed isolates to rapidly identify a cosmid clone which is tightly linked to the rad11 locus and is therefore a suitable starting point for a chromosome walk. These rapid methods of gene mapping and isolation should be applicable to any organism with separable chromosomes. Images PMID:1354851

  6. Amperometric sulfide detection using Coprinus cinereus peroxidase immobilized on screen printed electrode in an enzyme inhibition based biosensor.

    Science.gov (United States)

    Savizi, Iman Shahidi Pour; Kariminia, Hamid-Reza; Ghadiri, Mohammad; Roosta-Azad, Reza

    2012-05-15

    In the present work, an amperometric inhibition biosensor for the determination of sulfide has been fabricated by immobilizing Coprinus cinereus peroxidase (CIP) on the surface of screen printed electrode (SPE). Chitosan/acrylamide was applied for immobilization of peroxidase on the working electrode. The amperometric measurement was performed at an applied potential of -150 mV versus Ag/AgCl with a scan rate of 100 mV in the presence of hydroquinone as electron mediator and 0.1M phosphate buffer solution of pH 6.5. The variables influencing the performance of sensor including the amount of substrate, mediator concentration and electrolyte pH were optimized. The determination of sulfide can be achieved in a linear range of 1.09-16.3 μM with a detection limit of 0.3 μM. Developed sensor showed quicker response to sulfide compared to the previous developed sulfide biosensors. Common anions and cations in environmental water did not interfere with sulfide detection by the developed biosensor. Cyanide interference on the enzyme inhibition caused 43.25% error in the calibration assay which is less than the amounts reported by previous studies. Because of high sensitivity and the low-cost of SPE, this inhibition biosensor can be successfully used for analysis of environmental water samples. Copyright © 2012 Elsevier B.V. All rights reserved.

  7. A novel laccase with potent antiproliferative and HIV-1 reverse transcriptase inhibitory activities from mycelia of mushroom Coprinus comatus.

    Science.gov (United States)

    Zhao, Shuang; Rong, Cheng-Bo; Kong, Chang; Liu, Yu; Xu, Feng; Miao, Qian-Jiang; Wang, Shou-Xian; Wang, He-Xiang; Zhang, Guo-Qing

    2014-01-01

    A novel laccase was isolated and purified from fermentation mycelia of mushroom Coprinus comatus with an isolation procedure including three ion-exchange chromatography steps on DEAE-cellulose, CM-cellulose, and Q-Sepharose and one gel-filtration step by fast protein liquid chromatography on Superdex 75. The purified enzyme was a monomeric protein with a molecular weight of 64 kDa. It possessed a unique N-terminal amino acid sequence of AIGPVADLKV, which has considerably high sequence similarity with that of other fungal laccases, but is different from that of C. comatus laccases reported. The enzyme manifested an optimal pH value of 2.0 and an optimal temperature of 60°C using 2,2'-azinobis(3-ethylbenzothiazolone-6-sulfonic acid) diammonium salt (ABTS) as the substrate. The laccase displayed, at pH 2.0 and 37°C, K(m) values of 1.59 mM towards ABTS. It potently suppressed proliferation of tumor cell lines HepG2 and MCF7, and inhibited human immunodeficiency virus type 1 (HIV-1) reverse transcriptase (RT) with an IC50 value of 3.46 μM, 4.95 μM, and 5.85 μM, respectively, signifying that it is an antipathogenic protein.

  8. The Rad9 Gene of Coprinus Cinereus Encodes a Proline-Rich Protein Required for Meiotic Chromosome Condensation and Synapsis

    Science.gov (United States)

    Seitz, L. C.; Tang, K.; Cummings, W. J.; Zolan, M. E.

    1996-01-01

    The rad9 gene of Coprinus cinereus is essential for the normal completion of meiosis. We examined surface-spread preparations of wild-type and rad9-1 nuclei from the meiotic stages of karyogamy through metaphase I, and we determined the primary sequence, structure, and meiotic expression of the rad9 gene. In wild-type C. cinereus, karyogamy is followed by condensation and alignment of homologous chromosomes. Condensation and axial core development largely precede synapsis, which often initiates at telomeres. A diffuse diplotene phase coincides with dissolution of the synaptonemal complex, and subsequently chromosomes further condense as the cells progress into metaphase I. In contrast, although karyogamy and nucleolar fusion are apparently normal in rad9-1 basidia, only short stretches of synaptonemal complex form. These correlate with stretches of condensed chromatin, mostly at apparent chromosome ends, and regions of presumptive triple synapsis are numerous. rad9-1 basidia enter the diffuse stage of early diplotene, and then 50% of these cells enter metaphase I by the criteria of nucleolar elimination and at least some chromatin condensation. rad9 gene expression is induced after gamma irradiation and during meiosis. The gene has 27 exons and encodes a predicted protein of 2157 amino acids, with a proline-rich amino terminus. PMID:8846891

  9. A chimeric homeodomain protein causes self-compatibility and constitutive sexual development in the mushroom Coprinus cinereus.

    Science.gov (United States)

    Kües, U; Göttgens, B; Stratmann, R; Richardson, W V; O'Shea, S F; Casselton, L A

    1994-01-01

    The A mating type genes of the mushroom Coprinus cinereus encode two classes of putative transcription factor with distinctive homeodomain motifs (HD1 and HD2). A successful mating brings together different allelic forms of these genes and this triggers part of a developmental sequence required for sexual reproduction. In this report we provide evidence that this developmental programme is promoted by a physical interaction between the two classes of homeodomain protein. Rare dominant mutations conferring self-compatibility map to the A locus and result in constitutive operation of the A-regulated developmental pathway. Our molecular analysis of one of these mutations shows that it has generated a chimeric gene by inframe fusion of an HD2 and an HD1 gene. Fusion has overcome the normal incompatibility between two proteins coded by genes of the same A locus and generated a protein that is sufficient to promote development in the absence of any other active A mating type genes. The fusion protein retains most of the HD2 sequence, but only the C-terminal part of the HD1 protein. It has only the HD2 homeodomain motif as a potential DNA binding domain fused to an essential C-terminal region of the HD1 protein, which in a normal HD1-HD2 protein complex may be the major activation domain. Images PMID:7915674

  10. Three subfamilies of pheromone and receptor genes generate multiple B mating specificities in the mushroom Coprinus cinereus.

    Science.gov (United States)

    Halsall, J R; Milner, M J; Casselton, L A

    2000-01-01

    The B mating type locus of the basidiomycete Coprinus cinereus encodes a large family of lipopeptide pheromones and their seven transmembrane domain receptors. Here we show that the B42 locus, like the previously described B6 locus, derives its unique specificity from nine multiallelic genes that are organized into three subgroups each comprising a receptor and two pheromone genes. We show that the three genes within each group are kept together as a functional unit by being embedded in an allele-specific DNA sequence. Using a combination of sequence analysis, Southern blotting, and DNA-mediated transformation with cloned genes, we demonstrate that different B loci may share alleles of one or two groups of genes. This is consistent with the prediction that the three subgroups of genes are functionally redundant and that it is the different combinations of their alleles that generate the multiple B mating specificities found in nature. The B42 locus was found to contain an additional gene, mfs1, that encodes a putative multidrug transporter belonging to the major facilitator family. In strains with other B mating specificities, this gene, whose functional significance was not established, lies in a region of shared homology flanking the B locus. PMID:10757757

  11. Coprinuslactone protects the edible mushroom Coprinus comatus against biofilm infections by blocking both quorum-sensing and MurA.

    Science.gov (United States)

    de Carvalho, Maira P; Gulotta, Giuseppe; do Amaral, Matheus W; Lünsdorf, Heinrich; Sasse, Florenz; Abraham, Wolf-Rainer

    2016-11-01

    Pathogens embedded in biofilms are involved in many infections and are very difficult to treat with antibiotics because of higher resistance compared with planktonic cells. Therefore, new approaches for their control are urgently needed. One way to search for biofilm dispersing compounds is to look at defense strategies of organisms exposed to wet environments, which makes them prone to biofilm infections. It is reasonable to assume that mushrooms have developed mechanisms to control biofilms on their sporocarps (fruiting bodies). A preliminary screening for biofilms on sporocarps revealed several species with few or no bacteria on their sporocarps. From the edible mushroom Coprinus comatus where no bacteria on the sporocarp could be detected (3R,4S)-2-methylene-3,4-dihydroxypentanoic acid 1,4-lactone, named coprinuslactone, was isolated. Coprinuslactone interfered with quorum-sensing and dispersed biofilms of Pseudomonas aeruginosa, where it also reduced the formation of the pathogenicity factors pyocyanin and rhamnolipid B. Coprinuslactone also damaged Staphylococcus aureus cells in biofilms at subtoxic concentrations. Furthermore, it inhibited UDP-N-acetylglucosamine enolpyruvyl transferase (MurA), essential for bacterial cell wall synthesis. These two modes of action ensure the inhibition of a broad spectrum of pathogens on the fruiting body but may also be useful for future clinical applications. © 2016 Society for Applied Microbiology and John Wiley & Sons Ltd.

  12. Nutrients and non-nutrients composition and bioactivity of wild and cultivated Coprinus comatus (O.F.Müll.) Pers.

    Science.gov (United States)

    Stojković, Dejan; Reis, Filipa S; Barros, Lillian; Glamočlija, Jasmina; Ćirić, Ana; van Griensven, Leo J I D; Soković, Marina; Ferreira, Isabel C F R

    2013-09-01

    Mushrooms have been reported as sources of biomolecules with various potential. Coprinus comatus was studied to obtain information about this species, comparing cultivated and wild samples. Free sugars, fatty acids, tocopherols, organic acids and phenolic acids were analyzed by chromatographic techniques coupled to different detectors. C. comatus methanolic extract was tested for its antioxidant potential (reducing power, radical scavenging activity and lipid peroxidation inhibition) and antimicrobial properties (tested towards Gram positive and negative bacteria, and microfungi). The toxicity for liver cells was tested in porcine liver primary cells. Both studied samples revealed similar nutritional value and energy contribution. The cultivated sample revealed the highest content in free sugars, monounsaturated fatty acids and tocopherols, while the wild mushroom was richer in saturated and polyunsaturated fatty acids, organic acids and phenolic compounds. The cultivated species also revealed the highest antioxidant potential and antimicrobial activity (with exception towards Gram negative bacteria and Aspergillus ochraceus). Both species revealed no toxicity towards porcine liver cells. The present study proved that cultivated and wild mushrooms from the same species could be excellent options as food and as sources of nutritional and bioactive compounds. Furthermore, differences in wild and cultivated samples were comparatively investigated for the first time. Copyright © 2013 Elsevier Ltd. All rights reserved.

  13. Meiosis in Coprinus. VIII. A time-course study of the fusion and division of the spindle pole body during meiosis

    Science.gov (United States)

    1978-01-01

    The time-course study of meiosis in the fungus Coprinus cinereus (C. lagopus) by electron microscopy reveals that two monoglobular spindle pole bodies (SPB's) of prekaryogamy nuclei come together during karyogamy and are fused. The fusion SPB of postkaryogamy nucleus persists through zygotene and pachytene as evidenced by the presence of axial components and synaptonemal complexes. At early diplotene, the SPB divides. The divided SPB takes on a diglobular form, which grows in size to form two daughter SPB's. These separate and move to opposite poles at metaphase I. PMID:564915

  14. Mathematical modelling of morphogenesis in fungi: spatial organization of the gravitropic response in the mushroom stem of Coprinus cinereus.

    Science.gov (United States)

    Meskauskas, A; Moore, D; Novak Frazer, L

    1998-09-01

    The purpose of this work was to establish how the distribution of local curvatures changed during the mushroom stem gravitropic reaction and to suggest a suitable mathematical model based on these new data. The gravitropic bending of base- and apex-pinned Coprinus cinereus (Fries) S. F. Gray stems was recorded on videotapes. The images were captured from the tapes after each 10 min, rotated by 45 degrees and transformed into tables of changing co-ordinates of points for each stem. The non-linear regression of these points was performed using Legendre polynomials. From the resulting equations the patterns of changing local curvature for 50 subsections per stem during 400 min of gravitropic reaction were calculated. It was observed that base-pinned stems first bent from the apex, but later the curvature of this part decreased, and in the late stages the apex became nearly completely straight again. Subsections, located about one third of stem length from the base determined the main part of the final curvature. The free basal part of the apex-pinned stems bent upward and after a certain bending time also began to straighten. However, this process started significantly later and was weaker. Bending of the subsections close to the pinned apex did not stop when they reached the vertical position, and the final angle of gravitropic curvature could exceed 180 degrees. Plotting various functions of local bending speed and its derivatives against each other and against local angle indicated that, if the hypothetical signal about reorientation arises in the apex, its propagation towards the base did not follow simple wave or simple diffusion laws. The importance of the local angle of all subsections both for signal origin and transmission was established and a signal transmission equation, involving local angle of each subsection, was derived. After creating a suitable program this partial differential equation was solved numerically. The generated shapes of the bending stem

  15. Novel illudins from Coprinopsis episcopalis (syn. Coprinus episcopalis), and the distribution of illudin-like compounds among filamentous fungi.

    Science.gov (United States)

    Gonzalez del Val, Antonio; Platas, Gonzalo; Arenal, Francisco; Orihuela, Juan Carlos; Garcia, María; Hernández, Pilar; Royo, Inmaculada; De Pedro, Nuria; Silver, Lynn L; Young, Katherine; Vicente, María Francisca; Pelaez, Fernando

    2003-10-01

    The illudins are a family of fungal sesquiterpenes that have been studied as anti-tumor agents, and they also have antibacterial activity. Over a four-year period, 25 304 fungal isolates (approximately 97% ascomycetes and 3% basidiomycetes), were screened for antibacterial activity against methicillin-resistant Staphylococcus aureus. Illudin-like compounds with antibacterial and cytotoxic activity against tumor cell lines were observed in 10 basidiomycete strains. The isolates were recovered from different types of substrata using indirect methods and only formed sterile mycelia in pure culture. The isolates were genetically related but not identical, based on PCR-based fingerprinting techniques. DNA sequencing of the ITS1-5.8 S-ITS2 region of the strains revealed that nine had identical sequences, indicating that they were conspecific. The sequence of the remaining isolate was 96.34% similar, suggesting that it was a closely related species. The D1-D2 region of the 25 S rRNA gene of the two strain types was also sequenced. Both sequences were 99.39% similar, and Coprinopsis gonophylla (syn. Coprinus gonophyllus) was the closest match for both. Strains were grown in pure culture on a rice-based medium that allowed the development of basidiomata from one culture of the main strain type, which was identified as C. episcopalis, a close relative of C. gonophyllus. Both species (or strain types) produced different types of illudin-like compounds. Three novel illudins (I, I2 and J2) were found to be produced by the cultures identified as C. episcopalis, while only illudinic acid was produced by the other Coprinopsis sp. The taxonomical relationships of the Coprinops is species identified in this study with other illudin producers previously reported in the literature are discussed.

  16. Novel Ca2+/calmodulin-dependent protein kinase expressed in actively growing mycelia of the basidiomycetous mushroom Coprinus cinereus.

    Science.gov (United States)

    Kaneko, Keisuke; Yamada, Yusuke; Sueyoshi, Noriyuki; Watanabe, Akira; Asada, Yasuhiko; Kameshita, Isamu

    2009-01-01

    We isolated cDNA clones for novel protein kinases by expression screening of a cDNA library from the basidiomycetous mushroom Coprinus cinereus. One of the isolated clones was found to encode a calmodulin (CaM)-binding protein consisting of 488 amino acid residues with a predicted molecular weight of 53,906, which we designated CoPK12. The amino acid sequence of the catalytic domain of CoPK12 showed 46% identity with those of rat Ca2+/CaM-dependent protein kinase (CaMK) I and CaMKIV. However, a striking difference between these kinases is that the critical Thr residue in the activating phosphorylation site of CaMKI/IV is replaced by a Glu residue at the identical position in CoPK12. As predicted from its primary sequence, CoPK12 was found to behave like an activated form of CaMKI phosphorylated by an upstream CaMK kinase, indicating that CoPK12 is a unique CaMK with different properties from those of the well-characterized CaMKI, II, and IV. CoPK12 was abundantly expressed in actively growing mycelia and phosphorylated various proteins, including endogenous substrates, in the presence of Ca2+/CaM. Treatment of mycelia of C. cinereus with KN-93, which was found to inhibit CoPK12, resulted in a significant reduction in growth rate of mycelia. These results suggest that CoPK12 is a new type of multifunctional CaMK expressed in C. cinereus, and that it may play an important role in the mycelial growth.

  17. Sesquiterpene synthases Cop4 and Cop6 from Coprinus cinereus: Catalytic promiscuity and cyclization of farnesyl pyrophosphate geometrical isomers

    Science.gov (United States)

    Lopez-Gallego, Fernando; Agger, Sean A.; Pella, Daniel A.; Distefano, Mark D.; Schmidt-Dannert, Claudia

    2010-01-01

    Sesquiterpene synthases catalyze with different catalytic fidelity the cyclization of farnesyl pyrophosphate (FPP) into hundreds of known compounds with diverse structures and stereochemistries. Two sesquiterpene synthases, Cop4 and Cop6, were previously isolated from Coprinus cinereus as part of a fungal genome survey. This study investigates the reaction mechanism and catalytic fidelity of the two enzymes. Cyclization of all-trans-FPP ((E,E)-FPP) was compared to the cyclization of the cis-trans isomer of FPP ((Z,E)-FPP) as a surrogate for the secondary cisoid neryl cation intermediate generated by sesquiterpene synthases capable of isomerizing the C2-C3 π bond of all-trans-FPP. Cop6 is a “high-fidelity” α-cuprenene synthase that retains its fidelity under various conditions tested. Cop4 is a catalytically promiscuous enzyme that cyclizes (E,E)-FPP into multiple products, including (−)-germacrene D and cubebol. Changing the pH of the reaction drastically alters the fidelity of Cop4 and makes it a highly selective enzyme. Cyclization of (Z,E)-FPP by Cop4 and Cop6 yields products that are very different from those obtained with (E,E)-FPP. Conversion of (E,E)-FPP proceeds via a (6R)-β-bisabolyl carbocation in the case of Cop6 and an (E,E)-germacradienyl carbocation in the case of Cop4. However, (Z,E)-FPP is cyclized via a (6S)-β-bisabolene carbocation by both enzymes. Structural modeling suggests that differences in the active site and the loop that covers the active site of the two enzymes may explain their different catalytic fidelities. PMID:20419721

  18. The reversible depletion and reconstitution of a copper ion in Coprinus cinereus laccase followed by spectroscopic techniques.

    Science.gov (United States)

    Bukh, Christian; Bjerrum, Morten J

    2010-10-01

    The specific activities of crude and purified Coprinus cinereus laccase preparations could be enhanced by a factor of 10-12 by activation with copper ions. The copper to protein contents of purified non-activated laccase were 2.3+/-0.1 compared to 3.3+/-0.1 in purified activated laccase indicating that only a fraction of the laccase can be activated. Purified laccase not activated with copper ions shows in isoelectric focusing four bands in order of decreasing pI in a ratio 1/5/3/1 where only bands I and II had laccase activity. Purified activated laccase showed only three bands (I, II and III) in the ratio 5/4/1 all with some laccase activity. The pH profile of the activity for activated and non-activated laccase showed identical behavior indicating that the active forms were the same. The change in UV-Vis around 330 nm following the depletion and reconstitution of the enzyme combined with activity measurements supports the reversibility of the selective removal and insertion of copper ions at the type 2 site. The circular dichroism spectrum of activated purified laccase has characteristic changes around 350 nm relative to non-activated laccase indicative of changes at the type 2/type 3 sites. The difference between the electron paramagnetic resonance spectra of non-activated and activated C. cinereus laccase indicates that a fraction of the non-activated purified laccase contained a copper(II) signal with a coupling constant between a type 1 and a type 2 copper(II). This electron paramagnetic resonance signal could be explained by an induced asymmetry in the type 3 site due to a missing type 2 copper ion. Copyright 2010 Elsevier Inc. All rights reserved.

  19. Sesquiterpene synthases Cop4 and Cop6 from Coprinus cinereus: catalytic promiscuity and cyclization of farnesyl pyrophosphate geometric isomers.

    Science.gov (United States)

    Lopez-Gallego, Fernando; Agger, Sean A; Abate-Pella, Daniel; Distefano, Mark D; Schmidt-Dannert, Claudia

    2010-05-17

    Sesquiterpene synthases catalyze with different catalytic fidelity the cyclization of farnesyl pyrophosphate (FPP) into hundreds of known compounds with diverse structures and stereochemistries. Two sesquiterpene synthases, Cop4 and Cop6, were previously isolated from Coprinus cinereus as part of a fungal genome survey. This study investigates the reaction mechanism and catalytic fidelity of the two enzymes. Cyclization of all-trans-FPP ((E,E)-FPP) was compared to the cyclization of the cis-trans isomer of FPP ((Z,E)-FPP) as a surrogate for the secondary cisoid neryl cation intermediate generated by sesquiterpene synthases, which are capable of isomerizing the C2--C3 pi bond of all-trans-FPP. Cop6 is a "high-fidelity" alpha-cuprenene synthase that retains its fidelity under various conditions tested. Cop4 is a catalytically promiscuous enzyme that cyclizes (E,E)-FPP into multiple products, including (-)-germacrene D and cubebol. Changing the pH of the reaction drastically alters the fidelity of Cop4 and makes it a highly selective enzyme. Cyclization of (Z,E)-FPP by Cop4 and Cop6 yields products that are very different from those obtained with (E,E)-FPP. Conversion of (E,E)-FPP proceeds via a (6R)-beta-bisabolyl carbocation in the case of Cop6 and an (E,E)-germacradienyl carbocation in the case of Cop4. However, (Z,E)-FPP is cyclized via a (6S)-beta-bisabolene carbocation by both enzymes. Structural modeling suggests that differences in the active site and the loop that covers the active site of the two enzymes might explain their different catalytic fidelities.

  20. Frequent Changes in the Number of Reiterated Ribosomal RNA Genes Throughout the Life Cycle of the Basidiomycete Coprinus Cinereus

    Science.gov (United States)

    Pukkila, P. J.; Skrzynia, C.

    1993-01-01

    We have examined the stability of the tandemly repeated genes that encode the ribosomal RNA in Coprinus cinereus. These genes are contained within two linked HindIII fragments in a 3.0-Mb chromosome. We monitored the size of these fragments in both mitotic and meiotic segregants using the contour-clamped homogeneous electric field (CHEF) method. No length changes were observed in the smaller HindIII fragment (100 kb; 10 repeats) among the DNAs prepared from 46 asexual spore derivatives (oidia) or 128 meiotic segregants (basidiospores from 32 tetrads). However, the larger HindIII fragment (1100 kb; 120 repeats) did exhibit variability. Substantial changes, involving up to 40% of the larger HindIII fragment were recorded in 7 of 46 oidial isolates (including 4 of 22 transformed derivatives). To learn if the changes were confined to the vegetative portion of the life cycle, we examined transmission of HindIII variants through three crosses. In the first two crosses (16 tetrads total), no changes were observed in the large HindIII fragment. However, in the third cross (16 tetrads), each tetrad showed at least one alteration. In half of the tetrads from the third cross, the altered patterns segregated 2:2, suggesting that the changes occurred after mating but prior to premeiotic DNA replication. We conclude that breakage and rejoining reactions within the rDNA are frequent and are not confined to any particular stage of the life cycle. It also appears that certain repeats are sheltered from these events. Finally, marked differences in rDNA stability were observed in the crosses analyzed. PMID:8436270

  1. Molecular Characterization of Laccase Genes from the Basidiomycete Coprinus cinereus and Heterologous Expression of the Laccase Lcc1

    Science.gov (United States)

    Yaver, Debbie S.; Overjero, Maria Del Carmen; Xu, Feng; Nelson, Beth A.; Brown, Kim M.; Halkier, Torben; Bernauer, Sheryl; Brown, Stephen H.; Kauppinen, Sakari

    1999-01-01

    A laccase from Coprinus cinereus is active at alkaline pH, an essential property for some potential applications. We cloned and sequenced three laccase genes (lcc1, lcc2, and lcc3) from the ink cap basidiomycete C. cinereus. The lcc1 gene contained 7 introns, while both lcc2 and lcc3 contained 13 introns. The predicted mature proteins (Lcc1 to Lcc3) are 58 to 80% identical at the amino acid level. The predicted Lcc1 contains a 23-amino-acid C-terminal extension rich in arginine and lysine, suggesting that C-terminal processing may occur during its biosynthesis. We expressed the Lcc1 protein in Aspergillus oryzae and purified it. The Lcc1 protein as expressed in A. oryzae has an apparent molecular mass of 66 kDa on sodium dodecyl sulfate-polyacrylamide gel electrophoresis and absorption maxima at 278 and 614 nm. Based on the N-terminal protein sequence of the laccase, a 4-residue propeptide was processed during the maturation of the enzyme. The dioxygen specificity of the laccase showed an apparent Km of 21 ± 2 μM and a catalytic constant of 200 ± 10 min−1 for O2 with 2,2′-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) as the reducing substrate at pH 5.5. Lcc1 from A. oryzae may be useful in industrial applications. This is the first report of a basidiomycete laccase whose biosynthesis involves both N-terminal and C-terminal processing. PMID:10543807

  2. Involvement of Ca(2+)/calmodulin-dependent protein kinases in mycelial growth of the basidiomycetous mushroom, Coprinus cinereus.

    Science.gov (United States)

    Kameshita, Isamu; Yamada, Yusuke; Nishida, Tetsuyuki; Sugiyama, Yasunori; Sueyoshi, Noriyuki; Watanabe, Akira; Asada, Yasuhiko

    2007-09-01

    Although multifunctional Ca(2+)/calmodulin-dependent protein kinases (CaM-kinases) are widely distributed in animal cells, the occurrence of CaM-kinases in the basidiomycetous mushroom has not previously been documented. When the extracts from various developmental stages from mycelia to the mature fruiting body of Coprinus cinereus were analyzed by Western blotting using Multi-PK antibodies, which had been generated to detect a wide variety of protein serine/threonine kinases (Ser/Thr kinases), a variety of stage-specific Ser/Thr kinases was detected. Calmodulin (CaM) overlay assay using digoxigenin-labeled CaM detected protein bands of 65 kDa, 58 kDa, 46 kDa, 42 kDa, and 38 kDa only in the presence of CaCl(2), suggesting that these bands were CaM-binding proteins. When the CaM-binding fraction was prepared from mycelial extract of C. cinereus by CaM-Sepharose and analyzed with Multi-PK antibodies, two major immunoreactive bands corresponding to 65 kDa and 46 kDa were detected. CaM-binding fraction, thus obtained, exhibited Ca(2+)/CaM-dependent protein kinase activity toward protein substrates such as histones. These CaM-kinases were found to be highly expressed in the actively growing mycelia, but not in the resting mycelial cells. Mycelial growth was enhanced by the addition of CaCl(2) in the culture media, but inhibited by the addition of EGTA or trifluoperazine, a potent CaM inhibitor. This suggested that CaM-dependent enzymes including CaM-kinases play crucial roles in mycelial growth of basidiomycete C. cinereus.

  3. The clp1 gene of the mushroom Coprinus cinereus is essential for A-regulated sexual development.

    Science.gov (United States)

    Inada, K; Morimoto, Y; Arima, T; Murata, Y; Kamada, T

    2001-01-01

    Sexual development in the mushroom Coprinus cinereus is under the control of the A and B mating-type loci, both of which must be different for a compatible, dikaryotic mycelium to form between two parents. The A genes, encoding proteins with homeodomain motifs, regulate conjugate division of the two nuclei from each mating partner and promote the formation of clamp connections. The latter are hyphal configurations required for the maintenance of the nuclear status in the dikaryotic phase of basidiomycetes. The B genes encode pheromones and pheromone receptors. They regulate the cellular fusions that complete clamp connections during growth, as well as the nuclear migration required for dikaryosis. The AmutBmut strain (326) of C. cinereus, in which both A- and B-regulated pathways are constitutively activated by mutations, produces, without mating, dikaryon-like, fertile hyphae with clamp connections. In this study we isolated and characterized clampless1-1 (clp1-1), a mutation that blocks clamp formation, an essential step in A-regulated sexual development, in the AmutBmut background. A genomic DNA fragment that rescues the clp1-1 mutation was identified by transformations. Sequencing of the genomic DNA, together with RACE experiments, identified an ORF interrupted by one intron, encoding a novel protein of 365 amino acids. The clp1-1 mutant allele carries a deletion of four nucleotides, which is predicted to cause elimination of codon 128 and frameshifts thereafter. The clp1 transcript was normally detected only in the presence of the A protein heterodimer formed when homokaryons with compatible A genes were mated. Forced expression of clp1 by promoter replacements induced clamp development without the need for a compatible A gene combination. These results indicate that expression of clp1 is necessary and sufficient for induction of the A-regulated pathway that leads to clamp development. PMID:11139497

  4. Authentication Without Secrets

    Energy Technology Data Exchange (ETDEWEB)

    Pierson, Lyndon G. [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States); Robertson, Perry J. [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States)

    2015-11-01

    This work examines a new approach to authentication, which is the most fundamental security primitive that underpins all cyber security protections. Current Internet authentication techniques require the protection of one or more secret keys along with the integrity protection of the algorithms/computations designed to prove possession of the secret without actually revealing it. Protecting a secret requires physical barriers or encryption with yet another secret key. The reason to strive for "Authentication without Secret Keys" is that protecting secrets (even small ones only kept in a small corner of a component or device) is much harder than protecting the integrity of information that is not secret. Promising methods are examined for authentication of components, data, programs, network transactions, and/or individuals. The successful development of authentication without secret keys will enable far more tractable system security engineering for high exposure, high consequence systems by eliminating the need for brittle protection mechanisms to protect secret keys (such as are now protected in smart cards, etc.). This paper is a re-release of SAND2009-7032 with new figures numerous edits.

  5. Incretin secretion: direct mechanisms

    DEFF Research Database (Denmark)

    Balk-Møller, Emilie; Holst, Jens Juul; Kuhre, Rune Ehrenreich

    2014-01-01

    enzyme responsible for incretin degradation (dipeptidyl peptidase-4) is inhibited (drugs are already on the market) while the secretion of endogenous GLP-1 secretion is stimulated at the same time may prove particularly rewarding. In this section we review current knowledge on the mechanisms for direct......The incretin hormones glucose-dependent insulinotropic polypeptide (GIP) and glucagon-like peptide 1 (GLP-1) are secreted from gastro-intestinal K- and L-cells, respectively, and play an important role in post-prandial blood glucose regulation. They do this by direct stimulation of the pancreatic β...

  6. Immunoglobins in mammary secretions

    DEFF Research Database (Denmark)

    Hurley, W L; Theil, Peter Kappel

    2013-01-01

    Immunoglobulins secreted in colostrum and milk by the lactating mammal are major factors providing immune protection to the newborn. Immunoglobulins in mammary secretions represent the cumulative immune response of the lactating animal to exposure to antigenic stimulation that occurs through...... interaction with the environment. Extensive species variability exists in how and when maternal immunoglobulins are transferred to the neonate. In addition, there is a range of mechanisms by which the transferred immunoglobulins may play a protective role in the neonate. This chapter reviews...... the immunoglobulins found in mammary secretions in the context of their diversity of structure, origin, mechanisms of transfer, and function....

  7. Secreted proteases from dermatophytes.

    Science.gov (United States)

    Monod, Michel

    2008-01-01

    Dermatophytes are highly specialized pathogenic fungi that exclusively infect the stratum corneum, nails or hair, and it is evident that secreted proteolytic activity is important for their virulence. Endo- and exoproteases-secreted by dermatophytes are similar to those of species of the genus Aspergillus. However, in contrast to Aspergillus spp., dermatophyte-secreted endoproteases are multiple and are members of two large protein families, the subtilisins (serine proteases) and the fungalysins (metalloproteases). In addition, dermatophytes excrete sulphite as a reducing agent. In the presence of sulphite, disulphide bounds of the keratin substrate are directly cleaved to cysteine and S-sulphocysteine, and reduced proteins become accessible for further digestion by various endo- and exoproteases secreted by the fungi. Sulphitolysis is likely to be an essential step in the digestion of compact keratinized tissues which precedes the action of all proteases.

  8. A combined approach of experiments and computational docking simulation to the Coprinus cinereus peroxidase-catalyzed oxidative polymerization of alkyl phenols.

    Science.gov (United States)

    Park, Jong Chul; Joo, Jeong Chan; An, Eun Suk; Song, Bong Keun; Kim, Yong Hwan; Yoo, Young Je

    2011-04-01

    The characteristics of the oxidative polymerization of alkyl phenol derivatives catalyzed by Coprinus cinereus peroxidase (CIP) were studied qualitatively and quantitatively using a combined approach of experiments and computational docking simulations. As determined by docking study of CIP and alkyl phenols, the binding interaction was found to be important for the determination of substrate specificity. The distant binding and indirect orientation of o-isopropyl phenol and o-tertiary butyl phenol to the catalytic residue (56His) could explain the inability of CIP to polymerize these substrates. Three hydrophobic residues (156Pro, 192Leu, and 230Phe) at the entrance of the binding pocket were also found to be crucial in binding and orientation of alkyl phenols. A two-parameter QSAR equation with the binding distance and the molecular volume of the substrates was proposed and the polymerization yield was accurately predicted by two-parameter QSAR equation. Copyright © 2010 Elsevier Ltd. All rights reserved.

  9. Statistical Optimization of the Content Composition Precursors Using Response Surface Methodology to Enhance Agaricoglyceride A Production from the Shaggy Ink Cap Medicinal Mushroom, Coprinus comatus (Higher Basidiomycetes) Mycelia.

    Science.gov (United States)

    Wang, Wei; Di, Zhibiao; Li, Ruiguo; Tian, Jingzhen

    2015-01-01

    Coprinus comatus, a novel cultivated edible mushroom, has a various of pharmacological effects due to its many active components. In this study, agaricoglycerides, a new class of fungal secondary metabolites that have strong activity against neurolysin, were isolated from C. comatus mycelia. Simultaneously, a 3-level Box-Behnken factorial design was used, combined with response surface methodology, to optimize the precursor composition of agaricoglycerides for the production of agaricoglyceride A. The model estimated that a maximal yield of agaricoglyceride A (20.105 mg/L) could be obtained when the concentrations of 4-hydroxybenzoic acid, glycerol, and methanol (MeOH) were set at 75 mg/L, 0.75 mL/L, and 0.75 mL/L, respectively. The verified experiments showed that the model was significantly consistent with the model prediction. These results showed that appropriately adding the precursors could increase the production of agaricoglyceride A.

  10. Antineoplastic Agents 582. Isolation and Structure of a Cyclobutane-type Sesquiterpene Cancer Cell Growth Inhibitor from Coprinus cinereus (Coprinaceae)‡

    Science.gov (United States)

    Pettit, George R.; Meng, Yanhui; Pettit, Robin K.; Herald, Delbert L.; Hogan, Fiona; Cichacz, Zbigniew A.

    2010-01-01

    Bioassay-guided (murine P388 lymphocytic leukemia and human cancer cell lines) separation of an ethyl acetate extract prepared from the inky cap fungus Coprinus cinereus led to the isolation of three new sesquiterpenes, 7,7a-diepicoprinastatin 1 (1), 14-hydroxy-5-desoxy-2S,3S,9R-illudosin (2), and 4,5-dehydro-5-deoxyarmillol (3), together with the known armillol (4). The structure and relative configuration of 1 was determined by single-crystal X-ray diffraction experiments. The structures of compounds 2, 3, and 4 were each deduced by a combination of HRMS and 1D and 2D NMR techniques. Cyclobutane 2 led to modest inhibition of the murine P388 leukemia cell line. PMID:20598551

  11. Influence of nutritional supplementation on solid-substrate fermentation of wheat straw with an alkaliphilic white-rot fungus (Coprinus sp. )

    Energy Technology Data Exchange (ETDEWEB)

    Yadav, J.S.

    1987-08-01

    The solid-substrate fermentation of wheat straw with an alkaliphilic white-rot fungus (Coprinus sp.) was found to be influenced by the levels of nitrogen, phosphorus + sulphur and free carbohydrates, in terms of biodegradation of straw ingredients, microbial protein production and changes in in-vitro dry matter digestibility (IVDMD). Nitrogen and Phosphorus + Sulphur compounds favoured the bioconversion and their optimum levels were (g/100 g DM): urea (sterile): 1.5, urea (unsterile): 3.0; superphosphate: 1.0. The addition of free carbohydrates as molasses and whey had detrimental effect on biodegradation of lignin as also on organic matter degradation and digestibility. However, the protein production was enhanced in the supplemented straw. The optimized laboratory fermentation was also extended to 4 kg-(sterile and unsterile) and 50 kg-(unsterile) fermentation.

  12. Antineoplastic agents 582. Part 1: Isolation and structure of a cyclobutane-type sesquiterpene cancer cell growth inhibitor from Coprinus cinereus (Coprinaceae).

    Science.gov (United States)

    Pettit, George R; Meng, Yanhui; Pettit, Robin K; Herald, Delbert L; Hogan, Fiona; Cichacz, Zbigniew A

    2010-07-15

    Bioassay-guided (murine P388 lymphocytic leukemia and human cancer cell lines) separation of an ethyl acetate extract prepared from the inky cap fungus Coprinus cinereus led to the isolation of three new sesquiterpenes, 7,7a-diepicoprinastatin 1 (1), 14-hydroxy-5-desoxy-2S,3S,9R-illudosin (2), and 4,5-dehydro-5-deoxyarmillol (3), together with the known armillol (4). The structure and relative configuration of 1 was determined by single-crystal X-ray diffraction experiments. The structures of compounds 2, 3, and 4 were each deduced by a combination of HRMS and 1D and 2D NMR techniques. Cyclobutane 2 led to modest inhibition of the murine P388 leukemia cell line.

  13. Identification of a novel bifunctional delta12/delta15 fatty acid desaturase from a basidiomycete, Coprinus cinereus TD#822-2.

    Science.gov (United States)

    Zhang, Shuo; Sakuradani, Eiji; Ito, Kuni; Shimizu, Sakayu

    2007-01-23

    A new gene encoding a delta12 fatty acid desaturase-related protein was cloned from a multicellular basidiomycete Coprinus cinereus TD#822-2. The 1326 bp full-length gene, designated as Cop-odeA, codes for a putative protein of 442 amino acids with a MW of 49224. The Cop-odeA yeast transformant accumulated four new fatty acids identified as 9,12-hexadecadienoic acid, 9,12,15-hexadecatrienoic acid, linoleic acid, and alpha-linolenic acid, which comprised 8.8%, 1.0%, 29.0%, and 0.6% of the total fatty acids, respectively. The Cop-odeA protein was confirmed to be a novel bifunctional fatty acid desaturase with both high delta12 desaturase activity and unusual delta15 desaturase activity.

  14. IgE-binding proliferative responses and skin test reactivity to Cop c 1, the first recombinant allergen from the basidiomycete Coprinus comatus.

    Science.gov (United States)

    Brander, K A; Borbély, P; Crameri, R; Pichler, W J; Helbling, A

    1999-09-01

    Basidiomycetes spores are ubiquitously distributed, found throughout the year in outdoor and indoor air, and represent relevant sources of aeroallergens associated with allergy and asthma. Cloning and characterization of Coprinus comatus (shaggy cap mushroom) allergens is essential to elucidate their molecular characteristics and to improve the diagnosis of allergy. A complementary DNA (cDNA) library of C comatus displayed on phage surface was screened with sera of basidiomycete-sensitized individuals. Subcloning and high-level expression of one of the enriched cDNAs allowed the isolation of a [His](6)-tagged recombinant protein formally termed rCop c 1. The allergenic properties of rCop c 1 were investigated in vitro by ELISA, inhibition experiments, immunoblots, and proliferation assays and in vivo by skin tests. The rCop c 1-encoding cDNA spans 435 bp and contains an open reading frame of 246 bp, predicting a protein of 8.96 kd without significant sequence homology to known proteins. Immunoblots with [His](6)-rCop c 1 fusion protein show a background free IgE-binding band of the expected size that can be completely inhibited by crude C comatus extracts in ELISA. rCop c 1 induced specific proliferative responses in PBMCs of C comatus-sensitized individuals. The incidence of sensitization to rCop c 1 among 92 sera of basidiomycete-sensitized individuals tested in ELISA was 25%, indicating that Cop c 1 is an intermediate allergen. However, prick tests showed that less than 2 pmol of the rCop c 1 protein was able to induce strong specific skin reactions in sensitized individuals. rCop c 1, the first cloned allergen from the genus Coprinus, fulfills all the criteria required to be classified as a clinically relevant allergen. The data demonstrate at the molecular level the presence of sensitizing molecules among Basidiomycetes, the most important source contributing to the total spore load in the outdoor air.

  15. Computing on quantum shared secrets

    Science.gov (United States)

    Ouyang, Yingkai; Tan, Si-Hui; Zhao, Liming; Fitzsimons, Joseph F.

    2017-11-01

    A (k ,n )-threshold secret-sharing scheme allows for a string to be split into n shares in such a way that any subset of at least k shares suffices to recover the secret string, but such that any subset of at most k -1 shares contains no information about the secret. Quantum secret-sharing schemes extend this idea to the sharing of quantum states. Here we propose a method of performing computation securely on quantum shared secrets. We introduce a (n ,n )-quantum secret sharing scheme together with a set of algorithms that allow quantum circuits to be evaluated securely on the shared secret without the need to decode the secret. We consider a multipartite setting, with each participant holding a share of the secret. We show that if there exists at least one honest participant, no group of dishonest participants can recover any information about the shared secret, independent of their deviations from the algorithm.

  16. Salivary Gland Secretion.

    Science.gov (United States)

    Dorman, H. L.; And Others

    1981-01-01

    Describes materials and procedures for an experiment utilizing a live dog to demonstrate: (1) physiology of the salivary gland; (2) parasympathetic control of the salivary gland; (3) influence of varying salivary flow rates on sodium and potassium ions, osmolarity and pH; and (4) salivary secretion as an active process. (DS)

  17. A Public Secret

    DEFF Research Database (Denmark)

    Bregnbæk, Susanne

    2011-01-01

    This article is based on anthropological fieldwork undertaken at two elite universities in Beijing. It addresses the paradoxical situation of the many instances of suicide among Chinese elite university students in Beijing, which constitute a public secret. The pressure of education weighs heavily...

  18. Secrets of Successful Homeschooling

    Science.gov (United States)

    Rivero, Lisa

    2011-01-01

    Parents who homeschool gifted children often find the daily practice of home education very different from what they had imagined. Gifted children are complex in both personality and learning styles. Parents who say that homeschooling works well for their gifted children have learned from others or discovered on their own several secrets that make…

  19. Wrapped up in Covers: Preschoolers' Secrets and Secret Hiding Places

    Science.gov (United States)

    Corson, Kimberly; Colwell, Malinda J.; Bell, Nancy J.; Trejos-Castillo, Elizabeth

    2014-01-01

    In this qualitative study, interviews about children's secret hiding places were conducted with 3-5-year-olds (n?=?17) in a university sponsored preschool programme using art narratives. Since prior studies indicate that children understand the concept of a secret as early as five and that they associate secrets with hiding places, the purpose of…

  20. Extracellular secretion of recombinant proteins

    Science.gov (United States)

    Linger, Jeffrey G.; Darzins, Aldis

    2014-07-22

    Nucleic acids encoding secretion signals, expression vectors containing the nucleic acids, and host cells containing the expression vectors are disclosed. Also disclosed are polypeptides that contain the secretion signals and methods of producing polypeptides, including methods of directing the extracellular secretion of the polypeptides. Exemplary embodiments include cellulase proteins fused to secretion signals, methods to produce and isolate these polypeptides, and methods to degrade lignocellulosic biomass.

  1. Bucarest, Strictement Secret

    Directory of Open Access Journals (Sweden)

    Ionela Mihai

    2010-07-01

    Full Text Available L’émission Bucarest, strictement secret représente un documentaire organisésous la forme d’une série télé, qui dépeint le Bucarest à partir de deux perspectives: de l’histoire, de la conte et du lieu. La valeur d’une cité réside dans l’existence d’une mystique, d’un romantisme abscons, à part et des caractères empruntés de drames de Shakespeare, mystérieux, serrés d’angoisse et des secrets qui assombrissent leur existence. Par conséquence, le rôle du metteur en scène est de dévoiler leur vraie identité et de remettre en place, autant que possible, la vérité.

  2. Bile Formation and Secretion

    Science.gov (United States)

    Boyer, James L.

    2014-01-01

    Bile is a unique and vital aqueous secretion of the liver that is formed by the hepatocyte and modified down stream by absorptive and secretory properties of the bile duct epithelium. Approximately 5% of bile consists of organic and inorganic solutes of considerable complexity. The bile-secretory unit consists of a canalicular network which is formed by the apical membrane of adjacent hepatocytes and sealed by tight junctions. The bile canaliculi (~1 μm in diameter) conduct the flow of bile countercurrent to the direction of portal blood flow and connect with the canal of Hering and bile ducts which progressively increase in diameter and complexity prior to the entry of bile into the gallbladder, common bile duct, and intestine. Canalicular bile secretion is determined by both bile salt-dependent and independent transport systems which are localized at the apical membrane of the hepatocyte and largely consist of a series of adenosine triphosphate-binding cassette transport proteins that function as export pumps for bile salts and other organic solutes. These transporters create osmotic gradients within the bile canalicular lumen that provide the driving force for movement of fluid into the lumen via aquaporins. Species vary with respect to the relative amounts of bile salt-dependent and independent canalicular flow and cholangiocyte secretion which is highly regulated by hormones, second messengers, and signal transduction pathways. Most determinants of bile secretion are now characterized at the molecular level in animal models and in man. Genetic mutations serve to illuminate many of their functions. PMID:23897680

  3. The Secret Suburb

    DEFF Research Database (Denmark)

    Bech-Danielsen, Claus

    2015-01-01

    The ability to be ‘invisible’ seems to be an important quality in relation to a summerhouse. In fact, summerhouses can be said to be ‘invisible’ in a double sense. As I will explore in this chapter, summerhouses are neglected in planning and partly forgotten in Danish building regulations, at the......, at the same time as their owners like to see summerhouses as hidden places where they can live secret lives, hidden away from the modern world....

  4. Dynamic secrets in communication security

    CERN Document Server

    Xiao, Sheng; Towsley, Donald

    2013-01-01

    Dynamic secrets are constantly generated and updated from messages exchanged between two communication users. When dynamic secrets are used as a complement to existing secure communication systems, a stolen key or password can be quickly and automatically reverted to its secret status without disrupting communication. 'Dynamic Secrets in Communication Security' presents unique security properties and application studies for this technology. Password theft and key theft no longer pose serious security threats when parties frequently use dynamic secrets. This book also illustrates that a dynamic

  5. On Cheating Immune Secret Sharing

    Directory of Open Access Journals (Sweden)

    Josef Pieprzyk

    2004-12-01

    Full Text Available The paper addresses the cheating prevention in secret sharing. We consider secret sharing with binary shares. The secret also is binary. This model allows us to use results and constructions from the well developed theory of cryptographically strong boolean functions. In particular, we prove that for given secret sharing, the average cheating probability over all cheating vectors and all original vectors, i.e., 1/n 2 n ∑ c=1...n ∑ α∈V n ρ c,α, denoted by ρ, satisfies ρ ≥ ½, and the equality holds if and only if ρ c,α satisfies ρ c,α = ½ for every cheating vector δ c and every original vector α. In this case the secret sharing is said to be cheating immune. We further establish a relationship between cheating-immune secret sharing and cryptographic criteria of boolean functions.This enables us to construct cheating-immune secret sharing.

  6. Multiple Versions of the a Mating Type Locus of Coprinus Cinereus Are Generated by Three Paralogous Pairs of Multiallelic Homeobox Genes

    Science.gov (United States)

    Pardo, E. H.; O'Shea, S. F.; Casselton, L. A.

    1996-01-01

    The A mating type locus of Coprinus cinereus determines mating compatibility by regulating essential steps in sexual development. Each A locus contains several genes separated into two functionally independent complexes termed Aα and Aβ, and the multiple alleles of these genes generate an estimated 160 A mating specificities. The genes encode two classes of homeodomain-containing proteins designated HD1 and HD2. In this report we describe two newly cloned loci, A2 and A5, and compare them with A42, A43 and A6 that we have described previously. An Aβ-null locus, retaining just a single active HD1 gene from the α-complex, was generated by mutation. Using this as a transformation host, gene combinations that promote A-regulated development were identified. We demonstrate that each A locus contains members of three paralogous pairs of HD1 and HD2 genes. Different allelic versions of gene pairs are compatible but paralogous genes are incompatible. The genes present in four uncloned A loci were deduced using Southern analyses and transformations with available cloned genes. The combined analysis of nine A factors identifies sufficient A gene alleles to generate at least 72 A mating specificities. PMID:8878675

  7. A mutation in the FHA domain of Coprinus cinereus Nbs1 Leads to Spo11-independent meiotic recombination and chromosome segregation.

    Science.gov (United States)

    Crown, K Nicole; Savytskyy, Oleksandr P; Malik, Shehre-Banoo; Logsdon, John; Williams, R Scott; Tainer, John A; Zolan, Miriam E

    2013-11-06

    Nbs1, a core component of the Mre11-Rad50-Nbs1 complex, plays an essential role in the cellular response to DNA double-strand breaks (DSBs) and poorly understood roles in meiosis. We used the basidiomycete Coprinus cinereus to examine the meiotic roles of Nbs1. We identified the C. cinereus nbs1 gene and demonstrated that it corresponds to a complementation group previously known as rad3. One allele, nbs1-2, harbors a point mutation in the Nbs1 FHA domain and has a mild spore viability defect, increased frequency of meiosis I nondisjunction, and an altered crossover distribution. The nbs1-2 strain enters meiosis with increased levels of phosphorylated H2AX, which we hypothesize represent unrepaired DSBs formed during premeiotic replication. In nbs1-2, there is no apparent induction of Spo11-dependent DSBs during prophase. We propose that replication-dependent DSBs, resulting from defective replication fork protection and processing by the Mre11-Rad50-Nbs1 complex, are competent to form meiotic crossovers in C. cinereus, and that these crossovers lead to high levels of faithful chromosome segregation. In addition, although crossover distribution is altered in nbs1-2, the majority of crossovers were found in subtelomeric regions, as in wild-type. Therefore, the location of crossovers in C. cinereus is maintained when DSBs are induced via a Spo11-independent mechanism.

  8. Isolation and characterization of heavy-metal-mobilizing bacteria from contaminated soils and their potential in promoting Pb, Cu, and Cd accumulation by Coprinus comatus.

    Science.gov (United States)

    Jing, Xiao-Bing; He, Nan; Zhang, Ying; Cao, Yan-Ru; Xu, Heng

    2012-01-01

    The enhanced effect of heavy-metal-mobilizing bacteria on the uptake of Pb, Cu, and Cd by Coprinus comatus from Pb-, Cu-, and Cd-multicontaminated soil was assessed in this study. Thirteen strains, tolerating 800 mg·L(-1) Pb, 200 mg·L(-1) Cu, and 200 mg·L(-1) Cd simultaneously were selected for heavy-metal-solubilizing experiments in soil. The mobilization of heavy metals depended on the characteristics of bacteria and heavy metals. Correlation analysis demonstrated that for Pb solubilization, the acid-producing ability was the most significant factor, while for Cu and Cd, siderophores played a leading role in this process. Four strains, based on their excellent ability to solubilize heavy metal in soil, were applied in pot experiments. The results showed that all strains can promote the growth of C. comatus and meanwhile help mushroom accumulate more heavy metals (Pb, Cd, and Cu). The maximum uptake for total Pb and Cu by C. comatus was observed in inoculations with Bacillus sp. strain JSG1 (2.02- and 2.13-fold, respectively, compared with uninoculated soil), while for Cd, it was recorded in Bacillus sp. strain PB2 treated soil (2.03-fold). Therefore, this work suggests that the mushroom-bacteria interaction can be developed into a novel bioremediation strategy.

  9. Decolorization of synthetic dyes by crude and purified laccases from Coprinus comatus grown under different cultures: the role of major isoenzyme in dyes decolorization.

    Science.gov (United States)

    Jiang, Man; Ten, Zhen; Ding, Shaojun

    2013-01-01

    Coprinus comatus laccase isoenzyme induction and its effect on decolorization were investigated. The C/N ratio, together with aromatic compounds and copper, significantly influenced laccase isoenzyme profile and enzyme activity. This fungus produced six laccase isoenzymes in high-nitrogen low-carbon cultures but much less in low-nitrogen high-carbon (LNHC) cultures. The highest laccase level (3.25 IU/ml), equivalent to a 12.6-fold increase compared with unsupplemented controls (0.257 IU/ml), was recorded after 13 days in LNHC cultures supplemented with 2.0 mM 2-toluidine. Decolorization of twelve synthetic dyes belonging to anthraquinone, azo, and triphenylmethane dyes, by crude laccases with different proportion of isoenzymes produced under selected culture conditions, illustrated that the LacA is the key isoenzyme contributed to dyes decolorization especially in the presence of 1-hydroxybenzotriazol, which was further confirmed by dyes decolorization with purified LacA in the same condition. The crude laccase only was able to decolorize over 90 % of Reactive Brilliant Blue K-3R, Reactive Dark Blue KR, and Malachite Green, and higher decolorization for broader spectrum of synthetic dyes was obtained in presence of redox mediator, suggesting that C. comatus had high potential to decolorize various synthetic dyes as well as the recalcitrant azo dyes.

  10. The Shaggy Ink Cap Medicinal Mushroom, Coprinus comatus (Higher Basidiomycetes) Extract Induces Apoptosis in Ovarian Cancer Cells via Extrinsic and Intrinsic Apoptotic Pathways.

    Science.gov (United States)

    Rouhana-Toubi, Amal; Wasser, Solomon P; Fares, Fuad

    2015-01-01

    In a previous study, ethyl acetate extract of Coprinus comatus was found to reduce viability of human ovarian cancer cells. The objective of the current research was to clarify the mechanism of action of this extract. Ovarian cancer cells (ES-2) were subjected to ethyl acetate extract of C. comatus for different concentrations or exposure times. Cell cycle analysis and annexin V staining were performed using an automated flow cytometer. DNA fragmentation was detected using the TUNEL assay. Western blot analysis was performed for the assessment of activation of caspases -3, -8, and -9. Results revealed that treatment of ES-2 cells with ethyl acetate extract of C. comatus (100 μg/ml medium), for 48 h or for 72 h resulted in an increased number of cells at the sub-G1 phase of the cell cycle. These treatments also resulted in an increased number of apoptotic cells (positively stained by annexin and positively labeled by TUNEL), in comparison to the control. Reduced levels of procaspases -3, -8, and-9 were also detected in treated cells. In conclusion, ethyl acetate extract of C. comatus induces apoptosis in ovarian cancer cells (ES-2), via both extrinsic and intrinsic pathways. Meanwhile, more investigations are needed to demonstrate weather the apoptotic effect on ovarian cancer cells is accomplished by one active compound, or combined activities of different compounds that exist in the extract.

  11. Hypoglycemic Activity of Polysaccharide from Fruiting Bodies of the Shaggy Ink Cap Medicinal Mushroom, Coprinus comatus (Higher Basidiomycetes), on Mice Induced by Alloxan and Its Potential Mechanism.

    Science.gov (United States)

    Zhou, Shuai; Liu, Yanfang; Yang, Yan; Tang, Qingjiu; Zhang, Jingsong

    2015-01-01

    Three polysaccharide fractions from fruiting bodies of Coprinus comatus-CC30, CC60, and CC80-are obtained by water extraction and ethanol precipitation with ethanol percentages of 30%, 60%, and 80%, respectively. The hypoglycemic activity of the three fractions was tested based on mice with alloxan-induced diabetes. Results indicate that fraction CC60 is the most effective fraction in water extract from C. comatus; it can remarkably reduce the blood glucose concentration in 120 min at a dosage of 1000 mg/kg administered orally. It also presents a long-term hypoglycemic effect during 21 days of injection at the same dosage. This polysaccharide fraction provide a novel path to improve the treatment currently used for patients with diabetes. The data on mice spleen lymphocyte proliferation and protein tyrosine phosphatase 1B (PTP1B)-inhibiting activity of fractions indicate that the hypoglycemic activity of CC60 is possibly activated through immune stimulation, not PTP1B inhibition.

  12. The eln3 gene involved in fruiting body morphogenesis of Coprinus cinereus encodes a putative membrane protein with a general glycosyltransferase domain.

    Science.gov (United States)

    Arima, Toshihide; Yamamoto, Maki; Hirata, Aiko; Kawano, Shigeyuki; Kamada, Takashi

    2004-08-01

    We identified and characterized elongationless3 (eln3-1), a restriction enzyme-mediated integration (REMI) mutation affecting fruiting body morphogenesis in Coprinus cinereus. The mutant produces an aberrant fruiting body in which the stipe hardly elongates during fruiting body maturation. In the wild type, cylindrical stipe cells, elongation growth of which is responsible for stipe elongation, make side-by-side contact with one another and run parallel to the stipe axis, whereas in the mutant, the organization of the stipe tissue is disturbed and much space is produced between stipe cells. This disorganization of the stipe tissue, together with reduced elongation of the stipe cells, causes the mutant stipe short and bulgy. After a plasmid rescue, the eln3 gene was identified as a DNA fragment that complements the eln3-1 mutation. The eln3 ORF is predicted to encode a protein of 927 amino acids with a general glycosyltransferase domain and to be located in the plasma membrane. Transcription of the eln3 gene is specifically activated in rapidly elongating stipes. Possible involvement of the putative Eln3 enzyme in cell-to-cell connection is discussed.

  13. Roles of distal arginine in activity and stability of Coprinus cinereus peroxidase elucidated by kinetic and NMR analysis of the Arg51Gln, -Asn, -Leu, and -Lys mutants.

    Science.gov (United States)

    Schiødt, Christine B; Veitch, Nigel C; Welinder, Karen G

    2007-02-01

    In heme peroxidases, a distal His residue plays an essential role in the initial two electron oxidation of resting state enzyme to compound I by hydrogen peroxide. A distal Arg residue assists in this process. The contributions of the charge, H-bonding capacity, size, and mobility of this Arg residue to Coprinus cinereus peroxidase (CIP) reactivity and stability have been examined by substituting Arg51 with Gln (retains H-bond donor at N epsilon position), Asn (small size, H-bond donor and acceptor), Leu (similar to Asn, but hydrophobic), and Lys (charge and H-bond donor, but at N zeta position). UV-visible spectroscopy was used to monitor pH-linked heme changes, compound I formation and reduction, fluoride binding, and thermostability. (1)H NMR spectroscopy enabled heme pocket differences in both resting and cyanide-ligated states of the enzymes to be evaluated and compared with wild-type CIP. We found that the H-bonding capacity of distal Arg is key to fast compound I formation and ligand binding to heme, whereas charge is important for lowering the pK(a) of distal His and for the binding and stabilisation of anionic ligands at heme iron. The properties of the distal Arg residue in CIP, cytochrome c peroxidase (CCP) and horseradish peroxidase (HRP) differ significantly in their pH induced transitions and dynamics.

  14. Accumulation and tolerance to cadmium heavy metal ions and induction of 14-3-3 gene expression in response to cadmium exposure in Coprinus atramentarius.

    Science.gov (United States)

    Xie, Chengjian; Hu, Liujie; Yang, Yongzhu; Liao, Dunxiu; Yang, Xingyong

    2017-03-01

    Cadmium (Cd), one of the most toxic heavy-metal pollutants, has a strong and irreversible tendency to accumulate. Bioremediation is a promising technology to remedy and control heavy metal pollutants because of its low cost and ability to recycle heavy metals. Coprinus atramentarius is recognized as being able to accumulate heavy metal ions. In this work, C. atramentarius is cultivated on a solid medium containing Cd2+ ions to analyze its ability to tolerate different concentrations of the heavy metal ion. It is found that the growth of C. atramentarius is not significantly inhibited when the concentration of Cd2+ is less than 0.6mgL-1. The accumulation capacity of C. atramentarius at different Cd2+ concentrations also was determined. The results show that 76% of the Cd2+ present can be accumulated even when the concentration of the Cd2+ is 1mgL-1. The different proteins of C. atramentarius exposed to Cd2+ were further analyzed using gel electrophoresis. A 14-3-3 protein was identified and shown to be significantly up-regulated. In a further study, a full-length 14-3-3 gene was cloned containing a 759bp open reading frame encoding a polypeptide consisting of 252 amino acids and 3 introns. The gene expression work also showed that the 14-3-3 was significantly induced, and showed coordinated patterns of expression, with Cd2+ exposure. Copyright © 2016 Elsevier GmbH. All rights reserved.

  15. Inhibitory effect of ethyl acetate extract of the shaggy inc cap medicinal mushroom, Coprinus comatus (Higher Basidiomycetes) fruit bodies on cell growth of human ovarian cancer.

    Science.gov (United States)

    Rouhana-Toubi, Amal; Wasser, Solomon P; Agbarya, Abed; Fares, Fuad

    2013-01-01

    In a previous study, we screened organic extracts of different mushroom mycelia and picked ethyl acetate extract of Coprinus comatus as one of the most active extracts against human ovarian cancer cells. In the current study, we extracted a dry powder of C. comatus fruit bodies using ethyl acetate and examined its effect on the viability of three cell lines originated from human ovarian cancer (ES-2, SKOV-3, and SW-626). This extract was active against all tested cell lines, in a dose-dependent manner (concentrations 50-200 µg/mL, P<0.01). In an attempt to segregate the active fraction, we subjected the extract to chromatography on a silica gel column. The effect of six different fractions and of the crude extract on the viability of ES-2 cells was examined after exposure time of 24 h. Fraction F (last eluted) was significantly more effective than crude extract in the reduction of cell viability (P<0.01). Fraction F was also significantly more active than crude extract in the reduction of viability of SKOV-3 cells. We next identified some of the compounds of fraction F (mainly fatty acids) by gas chromatography-mass spectrometry. In summary, ethyl acetate extract of C. comatus reduced viability of three lines of human ovarian cancer. Fractionation of this extract by a silica gel column enabled the selection of a fraction significantly more active than the original extract.

  16. Optimization of cultivation conditions of fermented shaggy ink cap culinary-medicinal mushroom, Coprinus comatus (O.Mull.:Fr.) Pers. (higher Basidiomycetes) rich in Vanadium.

    Science.gov (United States)

    Zhang, Chunjing; Qi, Xiaodan; Shi, Yan; Sun, Yan; Li, Shuyan; Gao, Xiulan; Yu, Haitao

    2012-01-01

    The present paper is mainly aimed at optimization of cultivation conditions of fermented mushrooms of Coprinus comatus rich in vanadium (CCRV). Initial screening of effects of carbon source, temperature, pH, and inoculum size were done by using a one-factor-at-a-time method. The results obtained in that study showed that the optimal medium composition was 30 g glucose/Lin YEPG medium, initial pH 6.0, inoculum volume 10%, and incubation time 120 h. Then the medium was subjected to screening of the most significant parameters using the L9 orthogonal array to solve multivariable equations simultaneously. The results obtained in this study showed that the optimal medium composition was 0.4% V and 30 g glucose/Lin YEPG medium, initial pH 5.0, inoculum volume 15%, and incubation time 120 h. At this medium composition, the mycelial biomass and V content were 7.18 ± 0.24 g/L and 3786.0 ± 17 μg/g, respectively. The anti-diabetic potential of CCRV produced with the optimal level was tested in alloxan-induced diabetes. After the mice were administered (i.g.) with CCRV, the level of blood sugar in the CCRV group was very close to that of the control group. These findings suggested that CCRV produced with the optimal level is useful in the control of diabetes mellitus.

  17. Windows 8 secrets

    CERN Document Server

    Thurrott, Paul

    2012-01-01

    Tips, tricks, treats, and secrets revealed on Windows 8 Microsoft is introducing a major new release of its Windows operating system, Windows 8, and what better way to learn all its ins and outs than from two internationally recognized Windows experts and Microsoft insiders, authors Paul Thurrott and Rafael Rivera? They cut through the hype to get at useful information you'll not find anywhere else, including what role this new OS plays in a mobile and tablet world. Regardless of your level of knowledge, you'll discover little-known facts about how things work, what's new and different, and h

  18. Protecting Trade Secrets in Canada.

    Science.gov (United States)

    Courage, Noel; Calzavara, Janice

    2015-05-18

    Patents in the life sciences industries are a key form of intellectual property (IP), particularly for products such as brand-name drugs and medical devices. However, trade secrets can also be a useful tool for many types of innovations. In appropriate cases, trade secrets can offer long-term protection of IP for a lower financial cost than patenting. This type of protection must be approached with caution as there is little room for error when protecting a trade secret. Strong agreements and scrupulous security can help to protect the secret. Once a trade secret is disclosed to the public, it cannot be restored as the owner's property; however, if the information is kept from the public domain, the owner can have a property right of unlimited duration in the information. In some situations patents and trade secrets may be used cooperatively to protect innovation, particularly for manufacturing processes. Copyright © 2015 Cold Spring Harbor Laboratory Press; all rights reserved.

  19. Catecholamine Secretion from Individual Cells

    National Research Council Canada - National Science Library

    Wightman, R

    1998-01-01

    .... Many cells, including neurons, communicate by secretion of chemical substances by exocytosis where substances are extruded into the extracellular space following fusion of the vesicle and plasma membranes...

  20. Secret and research

    Directory of Open Access Journals (Sweden)

    André PETITAT

    2013-12-01

    Full Text Available The postures of secrecy and revelation maintain our common relational dynamics between sharing and not sharing. Science, which has become the dominant form of knowledge, is a rational and empirical knowledge sharing. For this purpose, the knowledge articulates languages, if possible unambiguous, spaces of rational deliberation, technical devices and resources of the imagination. This activity meets other logics called power, prestige, status, profit, customer, blind adherence and revealed truth, in which the postures of secret invite themselves massively. The codes of ethics attempt to regulate this mix of contradictory logics by setting standards of scientific exchanges, recalling the person rights and particularly the subjects observed rights, protecting the working conditions of the researcher, preserving its autonomy from funders and policy makers, and ensuring the dissemination of its results.

  1. Secret-key certificates (continued)

    NARCIS (Netherlands)

    S.A. Brands (Stefan)

    1995-01-01

    textabstractA new construction is described for designing secret-key certificate schemes based on signature schemes other than of the Fiat-Shamir type. Also described are practical secret-key certificate issuing protocols that enable the Certification Authority to certify public keys, without being

  2. A comparative overview of antioxidative properties and phenolic profiles of different fungal origins: fruiting bodies and submerged cultures of Coprinus comatus and Coprinellus truncorum.

    Science.gov (United States)

    Tešanović, Kristina; Pejin, Boris; Šibul, Filip; Matavulj, Milan; Rašeta, Milena; Janjušević, Ljiljana; Karaman, Maja

    2017-02-01

    Bioactive properties of fungi considerably differ between the fruiting body (FB) and the submerged culture as regards mycelia (M) and the fermentation broth (F). Antioxidant properties of hot-water extracts obtained from three different fungal origins: FB, M and F of two autochthonous fungal species (Northern Serbia), Coprinus comatus and Coprinellus truncorum were investigated. Free radical scavenging capacity (RSC) was evaluated in vitro by the DPPH assay and reducing power ability (FRAP assay). Considering possible bioactive properties of different compounds present in fungal extracts, the content of total proteins (TP), phenols (TC) and flavonoids (TF) were investigated colorimetrically. The chemical characterisation of the examined extracts was evaluated using the HPLC-MS/MS method. C. comatus showed the strongest RSC activity; more precisely, fermentation broth extract (FCc) on DPPH radicals (IC50 = 5.06 μg mL-1) and fruiting body extract (FBCc) for the FRAP assay (42.86 mg ascorbic acid equivalents (AAE)/g). Submerged M extract of both species showed the highest TC (MCc 81.95 mg gallic acid eq (GAE)/g d.w.; MCt 81.64 mg GAE/g d.w.), while FB extracts contained the highest content of TP. Comparing LC-MS phenolic profiles between species-interspecifically and among different fungal origins-intraspecifically (fruiting bodies and submerged cultures), high variations were noticed. In submerged M or F extracts of C. comatus, vanillic, gallic, gentisic and cinnamic acids were detected, as opposed to FB. Considering that diverse phenolic profiles of detected antioxidant compounds were obtained by submerged cultivation, this type of cultivation is promising for the production of antioxidant substances.

  3. Coprinus cinereus rad50 mutants reveal an essential structural role for Rad50 in axial element and synaptonemal complex formation, homolog pairing and meiotic recombination.

    Science.gov (United States)

    Acharya, Sonia N; Many, Alexander M; Schroeder, Andrew P; Kennedy, Felicia M; Savytskyy, Oleksandr P; Grubb, Jennifer T; Vincent, Jack A; Friedle, Elizabeth A; Celerin, Martina; Maillet, Daniel S; Palmerini, Heather J; Greischar, Megan A; Moncalian, Gabriel; Williams, R Scott; Tainer, John A; Zolan, Miriam E

    2008-12-01

    The Mre11/Rad50/Nbs1 (MRN) complex is required for eukaryotic DNA double-strand break (DSB) repair and meiotic recombination. We cloned the Coprinus cinereus rad50 gene and showed that it corresponds to the complementation group previously named rad12, identified mutations in 15 rad50 alleles, and mapped two of the mutations onto molecular models of Rad50 structure. We found that C. cinereus rad50 and mre11 mutants arrest in meiosis and that this arrest is Spo11 dependent. In addition, some rad50 alleles form inducible, Spo11-dependent Rad51 foci and therefore must be forming meiotic DSBs. Thus, we think it likely that arrest in both mre11-1 and the collection of rad50 mutants is the result of unrepaired or improperly processed DSBs in the genome and that Rad50 and Mre11 are dispensable in C. cinereus for DSB formation, but required for appropriate DSB processing. We found that the ability of rad50 mutant strains to form Rad51 foci correlates with their ability to promote synaptonemal complex formation and with levels of stable meiotic pairing and that partial pairing, recombination initiation, and synapsis occur in the absence of wild-type Rad50 catalytic domains. Examination of single- and double-mutant strains showed that a spo11 mutation that prevents DSB formation enhances axial element (AE) formation for rad50-4, an allele predicted to encode a protein with intact hook region and hook-proximal coiled coils, but not for rad50-1, an allele predicted to encode a severely truncated protein, or for rad50-5, which encodes a protein whose hook-proximal coiled-coil region is disrupted. Therefore, Rad50 has an essential structural role in the formation of AEs, separate from the DSB-processing activity of the MRN complex.

  4. Comparison of effects of vanadium absorbed by Coprinus comatus with those of inorganic vanadium on bone in streptozotocin-diabetic rats.

    Science.gov (United States)

    Wang, Guangbin; He, Ming; Yi, Pei; Wang, Jiashi; Li, Bin; Li, Jianjun; Fu, Yonghui; Bai, Lunhao; Fu, Qin

    2012-12-01

    The purpose of this study was to compare the effect of vanadium absorbed by Coprinus comatus (VACC) with inorganic vanadium (vanadium nitrate, IV) in preventing diabetes-related osteopenia in streptozotocin-diabetic rats. Sixty Wistar female rats used were divided into four groups: (1) normal rats (control), (2) diabetic rats, (3) diabetic rats treated with VACC, and (4) diabetic rats treated with vanadium nitrate. A standardized type 1-like diabetes model was induced by injection of streptozotocin. After the rats were treated orally with VACC and IV respectively, plasma glucose, body weights, micro-CT, biomechanical testing, and histomorphometry were examined. In addition, bone samples were obtained to evaluate the content of mineral substances in bones. Treatments were performed over a 12-week period. Both VACC and IV have a positive effect on plasma glucose and body weights of STZ-induced diabetic rats. However, treatment with IV only caused a 39.6 % decrease in glucose levels and a 14.6 % increase in body weights, whereas VACC decreased plasma glucose and increased body weights by up to 52.2 and 24.5 %, respectively. At the same time, VACC significantly improved trabecular microstructure and mechanical strength, while IV did not exhibit desirable such effects. Also, bone Ca and bone P were not significantly increased by IV. These results indicated that both VACC and IV have hypoglycemic activity on diabetic rats, while IV did not improve bone properties. In conclusion, this study suggests that VACC improves diabetes-related bone dysfunction, primarily by improving the diabetic states.

  5. Engineering the expression and characterization of two novel laccase isoenzymes from Coprinus comatus in Pichia pastoris by fusing an additional ten amino acids tag at N-terminus.

    Science.gov (United States)

    Gu, Chunjuan; Zheng, Fei; Long, Liangkun; Wang, Jing; Ding, Shaojun

    2014-01-01

    The detail understanding of physiological/biochemical characteristics of individual laccase isoenzymes in fungi is necessary for fundamental and application purposes, but our knowledge is still limited for most of fungi due to difficult to express laccases heterologously. In this study, two novel laccase genes, named lac3 and lac4, encoding proteins of 547 and 532-amino acids preceded by 28 and 16-residue signal peptides, respectively, were cloned from the edible basidiomycete Coprinus comatus. They showed 70% identity but much lower homology with other fungal laccases at protein level (less than 58%). Two novel laccase isoenzymes were successfully expressed in Pichia pastoris by fusing an additional 10 amino acids (Thr-Pro-Phe-Pro-Pro-Phe-Asn-Thr-Asn-Ser) tag at N-terminus, and the volumetric activities could be dramatically enhanced from undetectable level to 689 and 1465 IU/l for Lac3 and Lac4, respectively. Both laccases possessed the lowest Km and highest kcat/Km value towards syringaldazine, followed by ABTS, guaiacol and 2,6-dimethylphenol similar as the low redox potential laccases from other microorganisms. Lac3 and Lac4 showed resistant to SDS, and retained 31.86% and 43.08% activity in the presence of 100 mM SDS, respectively. Lac3 exhibited higher decolorization efficiency than Lac4 for eleven out of thirteen different dyes, which may attribute to the relatively higher catalytic efficiency of Lac3 than Lac4 (in terms of kcat/Km) towards syringaldazine and ABTS. The mild synergistic decolorization by two laccases was observed for triphenylmethane dyes but not for anthraquinone and azo dyes.

  6. Flutolanil and carboxin resistance in Coprinus cinereus conferred by a mutation in the cytochrome b560 subunit of succinate dehydrogenase complex (Complex II).

    Science.gov (United States)

    Ito, Y; Muraguchi, H; Seshime, Y; Oita, S; Yanagi, S O

    2004-10-01

    A gene that confers resistance to the systemic fungicide flutolanil was isolated from a mutant strain of the basidiomycete Coprinus cinereus. The flutolanil resistance gene was mapped to a chromosome of approximately 3.2 Mb, and a chromosome-specific cosmid library was constructed. Two cosmid clones that were able to transform a wild-type, flutolanil-sensitive, strain of C. cinereus to resistance were isolated from the library. Analysis of a subclone containing the resistance gene revealed the presence of the sdhC gene, which encodes the cytochrome b560 subunit of the succinate dehydrogenase (SDH) complex (Complex II) in the mitochondrial membrane. Comparison between the sdhC gene of a wild-type strain and that of a mutant strain revealed a single point mutation, which results in the replacement of Asn by Lys at position 80. Measurements of succinate-cytochrome c reductase activity in the transformants with mutant sdhC gene(s) suggest that flutolanil resistance of the fungus is caused by a decrease in the affinity of the SDH complex for flutolanil. This sdhC mutation also conferred cross-resistance against another systemic fungicide, carboxin, an anilide that is structurally related to flutolanil. In other organisms carboxin resistance mutations have been found in the genes sdhB and sdhD, but this is the first demonstration that a mutation in sdhC can also confer resistance. The mutant gene cloned in this work can be utilized as a dominant selectable marker in gene manipulation experiments in C. cinereus.

  7. Meiotic localization of Mre11 and Rad50 in wild type, spo11-1, and MRN complex mutants of Coprinus cinereus.

    Science.gov (United States)

    Many, Alexander M; Melki, Christina S; Savytskyy, Oleksandr P; Maillet, Daniel S; Acharya, Sonia N; Zolan, Miriam E

    2009-08-01

    The Mre11-Rad50-Nbs1 (MRN) complex is required for numerous cellular processes that involve interactions with DNA double-strand breaks. For the majority of these processes, the MRN complex is thought to act as a unit, with each protein aiding the activity of the others. We have examined the relationship between Mre11 and Rad50 during meiosis in the basidiomycete Coprinus cinereus (Coprinopsis cinerea), investigating to what extent activities of Mre11 and Rad50 are interdependent. We showed that mre11-1 is epistatic to rad50-1 with respect to the time of meiotic arrest, indicating that Mre11 activity facilitates the diffuse diplotene arrest of rad50 mutants. Anti-Mre11 and anti-Rad50 antibodies were used to examine MRN complex localization in a wild-type strain and in spo11, mre11, and rad50 mutants. In wild type, numbers of Mre11 and Rad50 foci peaked at time points corresponding to leptotene and early zygotene. In the spo11-1 mutant, which is defective in meiotic double-strand break formation, foci accumulated throughout prophase I. Of seven MRN mutants examined, only two rad50 strains exhibited Mre11 and Rad50 foci that localized to chromatin, although Mre11 protein was found in the cell for all of them. Analysis of predicted mutant structures showed that stable localization of Mre11 and Rad50 does not depend upon a wild-type hook-proximal coiled coil, but does require the presence of the Rad50 ATPase/adenylate cyclase domains. We found that Mre11 and Rad50 were interdependent for binding to meiotic chromosomes. However, the majority of foci observed apparently contained only one of the two proteins. Independent Mre11 and Rad50 foci might indicate disassociation of the complex during meiosis or could reflect independent structural roles for the two proteins in meiotic chromatin.

  8. Engineering the expression and characterization of two novel laccase isoenzymes from Coprinus comatus in Pichia pastoris by fusing an additional ten amino acids tag at N-terminus.

    Directory of Open Access Journals (Sweden)

    Chunjuan Gu

    Full Text Available The detail understanding of physiological/biochemical characteristics of individual laccase isoenzymes in fungi is necessary for fundamental and application purposes, but our knowledge is still limited for most of fungi due to difficult to express laccases heterologously. In this study, two novel laccase genes, named lac3 and lac4, encoding proteins of 547 and 532-amino acids preceded by 28 and 16-residue signal peptides, respectively, were cloned from the edible basidiomycete Coprinus comatus. They showed 70% identity but much lower homology with other fungal laccases at protein level (less than 58%. Two novel laccase isoenzymes were successfully expressed in Pichia pastoris by fusing an additional 10 amino acids (Thr-Pro-Phe-Pro-Pro-Phe-Asn-Thr-Asn-Ser tag at N-terminus, and the volumetric activities could be dramatically enhanced from undetectable level to 689 and 1465 IU/l for Lac3 and Lac4, respectively. Both laccases possessed the lowest Km and highest kcat/Km value towards syringaldazine, followed by ABTS, guaiacol and 2,6-dimethylphenol similar as the low redox potential laccases from other microorganisms. Lac3 and Lac4 showed resistant to SDS, and retained 31.86% and 43.08% activity in the presence of 100 mM SDS, respectively. Lac3 exhibited higher decolorization efficiency than Lac4 for eleven out of thirteen different dyes, which may attribute to the relatively higher catalytic efficiency of Lac3 than Lac4 (in terms of kcat/Km towards syringaldazine and ABTS. The mild synergistic decolorization by two laccases was observed for triphenylmethane dyes but not for anthraquinone and azo dyes.

  9. Pheochromocytomas and secreting paragangliomas

    Directory of Open Access Journals (Sweden)

    Gimenez-Roqueplo Anne-Paule

    2006-12-01

    Full Text Available Abstract Catecholamine-producing tumors may arise in the adrenal medulla (pheochromocytomas or in extraadrenal chromaffin cells (secreting paragangliomas. Their prevalence is about 0.1% in patients with hypertension and 4% in patients with a fortuitously discovered adrenal mass. An increase in the production of catecholamines causes symptoms (mainly headaches, palpitations and excess sweating and signs (mainly hypertension, weight loss and diabetes reflecting the effects of epinephrine and norepinephrine on α- and β-adrenergic receptors. Catecholamine-producing tumors mimic paroxysmal conditions with hypertension and/or cardiac rhythm disorders, including panic attacks, in which sympathetic activation linked to anxiety reproduces the same signs and symptoms. These tumors may be sporadic or part of any of several genetic diseases: familial pheochromocytoma-paraganglioma syndromes, multiple endocrine neoplasia type 2, neurofibromatosis 1 and von Hippel-Lindau disease. Familial cases are diagnosed earlier and are more frequently bilateral and recurring than sporadic cases. The most specific and sensitive diagnostic test for the tumor is the determination of plasma or urinary metanephrines. The tumor can be located by computed tomography, magnetic resonance imaging and metaiodobenzylguanidine scintigraphy. Treatment requires resection of the tumor, generally by laparoscopic surgery. About 10% of tumors are malignant either at first operation or during follow-up, malignancy being diagnosed by the presence of lymph node, visceral or bone metastases. Recurrences and malignancy are more frequent in cases with large or extraadrenal tumors. Patients, especially those with familial or extraadrenal tumors, should be followed-up indefinitely.

  10. Adrenal gland hormone secretion (image)

    Science.gov (United States)

    The adrenal gland secretes steroid hormones such as cortisol and aldosterone. It also makes precursors that can be converted ... steroids (androgen, estrogen). A different part of the adrenal gland makes adrenaline (epinephrine). When the glands produce ...

  11. VICTORIA'S SECRET Prepares for Growth

    National Research Council Canada - National Science Library

    Jordan K Speer

    2007-01-01

      After the upcoming holiday season, Victoria's Secret will take the first steps toward launching its e-commerce business on a cross-channel on-demand platform from partner n2N Commerce, a company...

  12. Of Plants, and Other Secrets

    Directory of Open Access Journals (Sweden)

    Michael Marder

    2012-12-01

    Full Text Available In this article, I inquire into the reasons for the all-too-frequent association of plants and secrets. Among various hypotheses explaining this connection from the standpoint of plant morphology and physiology, the one that stands out is the idea that plants are not only objects in the natural environment, but also subjects with a peculiar mode of accessing the world. The core of the “plant enigma” is, therefore, onto-phenomenological. Positively understood, the secret of their subjectivity leaves just enough space for the self-expression and the self-interpretation of vegetal life.

  13. Histaminergic regulation of prolactin secretion

    DEFF Research Database (Denmark)

    Knigge, U P

    1990-01-01

    Histamine (HA), which acts as a neurotransmitter in the central nervous system, participates in the neuroendocrine regulation of prolactin (PRL) secretion. HA has a predominant stimulatory effect which is mediated via H2-receptors following central administration and via H1-receptors following...

  14. Raspberry Pi for secret agents

    CERN Document Server

    Sjogelid, Stefan

    2015-01-01

    This book is an easy-to-follow guide with practical examples in each chapter. Suitable for the novice and expert alike, each topic provides a fast and easy way to get started with exciting applications and also guides you through setting up the Raspberry Pi as a secret agent toolbox.

  15. Stimulation of incretin secreting cells.

    Science.gov (United States)

    Pais, Ramona; Gribble, Fiona M; Reimann, Frank

    2016-02-01

    The incretin hormones glucose-dependent insulinotropic polypeptide (GIP) and glucagon like peptide-1 (GLP-1) are secreted from enteroendocrine cells in the gut and regulate physiological and homeostatic functions related to glucose control, metabolism and food intake. This review provides a systematic summary of the molecular mechanisms underlying secretion from incretin cells, and an understanding of how they sense and interact with lumen and vascular factors and the enteric nervous system through transporters and G-protein coupled receptors (GPCRs) present on their surface to ultimately culminate in hormone release. Some of the molecules described below such as sodium coupled glucose transporter 1 (SGLT1), G-protein coupled receptor (GPR) 119 and GPR40 are targets of novel therapeutics designed to enhance endogenous gut hormone release. Synthetic ligands at these receptors aimed at treating obesity and type 2 diabetes are currently under investigation.

  16. [Insulin secretion: mechanisms of regulation].

    Science.gov (United States)

    Radosavljević, Tatjana; Todorović, Vera; Sikić, Branka

    2004-01-01

    REGULATION OF INSULIN SECRETION: Beta cells are unique endocrine cells. They respond positively, in terms of insulin secretion, not only to changes in the extracellular glucose concentration, but also to activators of the phospholipase C (cholecystokinin or acetylcholine), and to activators of adenylate cyclase (glucagon, glucagon-like peptide-1, or gastric inhibitory polypeptide). Major messengers which mediate glucose action for insulin release are Ca2+, adenosine triphosphate (ATP) and diacylglycerol (DAG). MAJOR PATHWAYS OF INSULIN RELEASE STIMULATION: There are four major pathways involved in stimulation of insulin release. The first pathway is KATP channel-dependent pathway in which increased blood glucose concentrations and increased b-cell metabolism result in a change in intracellular ATP/ADP ratio. This is a contributory factor in closure of ATP-dependent K+ channels, depolarization of b-cell membrane, in increased voltage-dependent L-type Ca2+ channel activity. Increased Ca2+ influx results in increased intracellular Ca2+ and stimulated insulin release. KATP channel-independent pathway augments Ca(2+) -stimulated insulin secretion of KATP channel-dependent pathway. Major potentiation of release results from hormonal and peptidergic activation of receptors linked to adenylyl cyclase. Adenylyl cyclase activity is stimulated by hormones such as vasoactive intestinal peptide (VIP), glucagon-like peptide-1 (GLP-1), and so on. These hormones, acting via G protein, stimulate adenylyl cyclase, thus causing a rise in cyclic adenosine monophosphate (cAMP) and activation of protein kinase A (PKA). Increased activity of PKA results in potentiation of insulin secretion.

  17. Partager ses secrets en public

    OpenAIRE

    Merzeau, Louise

    2013-01-01

    National audience; We can be tempted to interpret the evolution of the Web and especially of social networks as a victory of an ideology of visibility, denying the right to withhold, withdraw and disconnect. Does this dictatorship of transparency, which is threatening even State secrets, means the end opacities, gray areas and moving frontiers between what is shown et whant is hidden ? We want to show that the digital environment is characterized instead by a regime of ambivalent visibility, ...

  18. Sec-mediated secretion by Coxiella burnetii

    Science.gov (United States)

    2013-01-01

    Background Coxiella burnetii is a Gram-negative intracellular bacterial pathogen that replicates within a phagolysosome-like parasitophorous vacuole (PV) of macrophages. PV formation requires delivery of effector proteins directly into the host cell cytoplasm by a type IVB secretion system. However, additional secretion systems are likely responsible for modification of the PV lumen microenvironment that promote pathogen replication. Results To assess the potential of C. burnetii to secrete proteins into the PV, we analyzed the protein content of modified acidified citrate cysteine medium for the presence of C. burnetii proteins following axenic (host cell-free) growth. Mass spectrometry generated a list of 105 C. burnetii proteins that could be secreted. Based on bioinformatic analysis, 55 proteins were selected for further study by expressing them in C. burnetii with a C-terminal 3xFLAG-tag. Secretion of 27 proteins by C. burnetii transformants was confirmed by immunoblotting culture supernatants. Tagged proteins expressed by C. burnetii transformants were also found in the soluble fraction of infected Vero cells, indicating secretion occurs ex vivo. All secreted proteins contained a signal sequence, and deletion of this sequence from selected proteins abolished secretion. These data indicate protein secretion initially requires translocation across the inner-membrane into the periplasm via the activity of the Sec translocase. Conclusions C. burnetii secretes multiple proteins, in vitro and ex vivo, in a Sec-dependent manner. Possible roles for secreted proteins and secretion mechanisms are discussed. PMID:24093460

  19. Transporter-mediated biofuel secretion.

    Science.gov (United States)

    Doshi, Rupak; Nguyen, Tuan; Chang, Geoffrey

    2013-05-07

    Engineering microorganisms to produce biofuels is currently among the most promising strategies in renewable energy. However, harvesting these organisms for extracting biofuels is energy- and cost-intensive, limiting the commercial feasibility of large-scale production. Here, we demonstrate the use of a class of transport proteins of pharmacological interest to circumvent the need to harvest biomass during biofuel production. We show that membrane-embedded transporters, better known to efflux lipids and drugs, can be used to mediate the secretion of intracellularly synthesized model isoprenoid biofuel compounds to the extracellular milieu. Transporter-mediated biofuel secretion sustainably maintained an approximate three- to fivefold boost in biofuel production in our Escherichia coli test system. Because the transporters used in this study belong to the ubiquitous ATP-binding cassette protein family, we propose their use as "plug-and-play" biofuel-secreting systems in a variety of bacteria, cyanobacteria, diatoms, yeast, and algae used for biofuel production. This investigation showcases the potential of expressing desired membrane transport proteins in cell factories to achieve the export or import of substances of economic, environmental, or therapeutic importance.

  20. QUANTITATIVE STUDIES OF PROSTATIC SECRETION

    Science.gov (United States)

    Huggins, Charles; Clark, Philip Johnson

    1940-01-01

    Cystic hyperplasia of the prostate occurs spontaneously in senile dogs only when they possess physiologically effective amounts of androgenic hormone. The cysts are closely grouped and radially arranged in a conical manner with the base of the cone at the periphery of the gland. Flattened and columnar epithelium, varying from about 5 to 25µ are seen in each cyst. The cysts communicate with the urethra by way of ducts. Both normal and cystic prostates undergo marked atrophy when the testes are removed, the chief difference 3 months after orchiectomy being the persistence of slightly dilated clefts and spaces at the site of the former cysts in the senile state. In the castrate dog whose prostate gland is being reconstructed as result of the influence of daily injections of androgen, certain doses of estrogen prevent increase of secretion and still larger doses greatly depress the output of the gland. In dogs so treated by daily injections of testosterone propionate, 10 mg., the amount of secretion is maintained from day to day at a level by daily injections of stilbestrol, 0.4 to 0.6 mg. and greatly depressed by doses of 1 to 1.5 mg. When the larger amounts of estrogen are used, together with androgen, squamous metaplasia occurs in the posterior lobe of the prostate while the epithelium of the acini decreases in height to cuboidal or low columnar form; these histological signs of activity of both androgen and estrogen on the prostate show that inhibition of the male hormone by stilbestrol is incomplete at these ratios. In dogs with either normal or cystic prostate glands, the prostate decreases in size when estrogen is injected in amounts to depress prostatic secretion profoundly. The gland is maintained in an atrophic state and overdosage avoided by controlled periodic injections of stilbestrol until secretion is reduced to the minimum, followed by free intervals, the estrogen being again administered when secretion measurably increases. The shrinkage is related to

  1. Incretin hormone secretion over the day

    DEFF Research Database (Denmark)

    Ahren, B; Carr, RD; Deacon, Carolyn F.

    2010-01-01

    . Regulation of incretin hormone secretion is less well characterized. The main stimulus for incretin hormone secretion is presence of nutrients in the intestinal lumen, and carbohydrate, fat as well as protein all have the capacity to stimulate GIP and GLP-1 secretion. More recently, it has been established...... that a diurnal regulation exists with incretin hormone secretion to an identical meal being greater when the meal is served in the morning compared to in the afternoon. Finally, whether incretin hormone secretion is altered in disease states is an area with, so far, controversial results in different studies......, although some studies have demonstrated reduced incretin hormone secretion in type 2 diabetes. This review summarizes our knowledge on regulation of incretin hormone secretion and its potential changes in disease states....

  2. LcrG secretion is not required for blocking of Yops secretion in Yersinia pestis

    Directory of Open Access Journals (Sweden)

    Matson Jyl S

    2008-02-01

    Full Text Available Abstract Background LcrG, a negative regulator of the Yersinia type III secretion apparatus has been shown to be primarily a cytoplasmic protein, but is secreted at least in Y. pestis. LcrG secretion has not been functionally analyzed and the relevance of LcrG secretion on LcrG function is unknown. Results An LcrG-GAL4AD chimera, originally constructed for two-hybrid analyses to analyze LcrG protein interactions, appeared to be not secreted but the LcrG-GAL4AD chimera retained the ability to regulate Yops secretion. This result led to further investigation to determine the significance of LcrG secretion on LcrG function. Additional analyses including deletion and substitution mutations of amino acids 2–6 in the N-terminus of LcrG were constructed to analyze LcrG secretion and LcrG's ability to control secretion. Some changes to the N-terminus of LcrG were found to not affect LcrG's secretion or LcrG's secretion-controlling activity. However, substitution of poly-isoleucine in the N-terminus of LcrG did eliminate LcrG secretion but did not affect LcrG's secretion controlling activity. Conclusion These results indicate that secretion of LcrG, while observable and T3SS mediated, is not relevant for LcrG's ability to control secretion.

  3. Weegee’s City Secrets

    Directory of Open Access Journals (Sweden)

    Alan TRACHTENBERG

    2010-03-01

    Full Text Available En tant que photographe indépendant de meurtres, d’accidents, d’incendies, mais aussi de moments de loisirs dans la ville — de scènes de violence et de plaisir — Weegee travaillait essentiellement la nuit et utilisait un flash puissant associé à son appareil-photo de presse. Ses « secrets pour réaliser des photographies avec un flash » consistent à donner des conseils pratiques et techniques pour débutants. Mais au cœur de la rhétorique de ses « secrets » se trouvent des réflexions subtiles et convaincantes révélant la relation entre la lumière et l’obscurité, et plus particulièrement la manière dont la lumière du flash permet de rendre visible l’obscurité. Dans le récit de Weegee, le flash confère à la photographie le pouvoir d’écrire — d’écrire avec la lumière, un mode de représentation singulièrement approprié pour enregistrer des instants de vie dans les rues nocturnes de la ville.As a freelance photographer of crime, accidents, fires, and also of the recreational life of the city—scenes of violence and of pleasure—Weegee worked mainly at night and employed a powerful photoflash attachment to his press camera. His "secrets of shooting with photoflash" consist of practical technical advice for beginners. But within the rhetoric of his "secrets" there lie cogent and subtle reflections on the relation of light to darkness, especially on the way the flash of light makes darkness visible. In Weegee’s account, the photoflash gives photography the power of writing—writing with light, a mode of picturing uniquely suited to recording instants of life on city streets at night.

  4. Britain's nuclear secrets: inside Sellafield

    Science.gov (United States)

    Marino, Antigone

    2017-11-01

    Lying on the remote north west coast of England, Sellafield is one of the most secret places in UK, and even one of the most controversial nuclear fuel reprocessing and nuclear decommissioning sites in Britain. The film director Tim Usborne let us enter into the world's first nuclear power station, revealing Britain's attempts to harness the almost limitless power of the atom. It is precisely the simplicity and the scientific rigor used in the film to speak of nuclear, which led this documentary to win the Physics Prize supported by the European Physical Society at the European Science TV and New Media Festival and Awards 2016.

  5. Secretion management in the mechanically ventilated patient.

    Science.gov (United States)

    Branson, Richard D

    2007-10-01

    Secretion management in the mechanically ventilated patient includes routine methods for maintaining mucociliary function, as well as techniques for secretion removal. Humidification, mobilization of the patient, and airway suctioning are all routine procedures for managing secretions in the ventilated patient. Early ambulation of the post-surgical patient and routine turning of the ventilated patient are common secretion-management techniques that have little supporting evidence of efficacy. Humidification is a standard of care and a requisite for secretion management. Both active and passive humidification can be used. The humidifier selected and the level of humidification required depend on the patient's condition and the expected duration of intubation. In patients with thick, copious secretions, heated humidification is superior to a heat and moisture exchanger. Airway suctioning is the most important secretion removal technique. Open-circuit and closed-circuit suctioning have similar efficacy. Instilling saline prior to suctioning, to thin the secretions or stimulate a cough, is not supported by the literature. Adequate humidification and as-needed suctioning are the foundation of secretion management in the mechanically ventilated patient. Intermittent therapy for secretion removal includes techniques either to simulate a cough, to mechanically loosen secretions, or both. Patient positioning for secretion drainage is also widely used. Percussion and postural drainage have been widely employed for mechanically ventilated patients but have not been shown to reduce ventilator-associated pneumonia or atelectasis. Manual hyperinflation and insufflation-exsufflation, which attempt to improve secretion removal by simulating a cough, have been described in mechanically ventilated patients, but neither has been studied sufficiently to support routine use. Continuous lateral rotation with a specialized bed reduces atelectasis in some patients, but has not been shown

  6. Accurate prediction of secreted substrates and identification of a conserved putative secretion signal for type III secretion systems.

    Directory of Open Access Journals (Sweden)

    Ram Samudrala

    2009-04-01

    Full Text Available The type III secretion system is an essential component for virulence in many Gram-negative bacteria. Though components of the secretion system apparatus are conserved, its substrates--effector proteins--are not. We have used a novel computational approach to confidently identify new secreted effectors by integrating protein sequence-based features, including evolutionary measures such as the pattern of homologs in a range of other organisms, G+C content, amino acid composition, and the N-terminal 30 residues of the protein sequence. The method was trained on known effectors from the plant pathogen Pseudomonas syringae and validated on a set of effectors from the animal pathogen Salmonella enterica serovar Typhimurium (S. Typhimurium after eliminating effectors with detectable sequence similarity. We show that this approach can predict known secreted effectors with high specificity and sensitivity. Furthermore, by considering a large set of effectors from multiple organisms, we computationally identify a common putative secretion signal in the N-terminal 20 residues of secreted effectors. This signal can be used to discriminate 46 out of 68 total known effectors from both organisms, suggesting that it is a real, shared signal applicable to many type III secreted effectors. We use the method to make novel predictions of secreted effectors in S. Typhimurium, some of which have been experimentally validated. We also apply the method to predict secreted effectors in the genetically intractable human pathogen Chlamydia trachomatis, identifying the majority of known secreted proteins in addition to providing a number of novel predictions. This approach provides a new way to identify secreted effectors in a broad range of pathogenic bacteria for further experimental characterization and provides insight into the nature of the type III secretion signal.

  7. Secreted Reporters for Monitoring Multiple Promoter Function.

    Science.gov (United States)

    Lashgari, Ghazal; Kantar, Rami S; Tannous, Bakhos A

    2017-01-01

    Secreted reporter proteins are reliable modalities for monitoring of different biological processes, which can be measured longitudinally in conditioned medium of cultured cells or body fluids such as blood and urine, ex vivo. In this chapter, we will explore established secreted reporters and their applications and limitations for monitoring of promoter function. We will also describe both cell-based and blood-based assays for detecting three commonly used reporters: secreted alkaline phosphatase (SEAP ), Gaussia luciferase (Gluc), and Vargula luciferase (Vluc).

  8. Secreted HSP Vaccine for Malaria Prophylaxis

    Science.gov (United States)

    2017-10-01

    AWARD NUMBER: W81XWH-13-2-0098 TITLE: Secreted HSP Vaccine for Malaria Prophylaxis PRINCIPAL INVESTIGATOR: Natasa Strbo CONTRACTING...1. REPORT DATE October 2017 2. REPORT TYPE Annual 3. DATES COVERED 09/30/16-09/29/17 4. TITLE AND SUBTITLE Secreted HSP Vaccine for Malaria ...thereby stimulating an avid, antigen specific, cytotoxic CD8 T cell response. Here we developed malaria vaccine that relies on secreted gp96-Ig

  9. Gene design, expression, crystallization and preliminary diffraction analysis of two isolectins from the fungus Coprinus cinereus: a model for studying functional diversification of galectins in the same organism and their evolutionary pathways.

    Science.gov (United States)

    López-Lucendo, María F; Giménez-Gallego, Guillermo; Cooper, Douglas N W; Gabius, Hans Joachim; Romero, Antonio

    2004-04-01

    It is the aim of comparative structural biology to define the evolutionarily important traits of protein function and the points of diversification. Consequently, structural analysis, especially of distant members in a family which in this case are lectins involved in cell adhesion and growth regulation in animals (i.e. galectins), is required. For this purpose, recent work has been focused on the first galectins known from outside the animal kingdom. These are the two isolectins from the basidiomycete Coprinus cinereus (inky cap mushroom), termed Cgl-1 and Cgl-2. Additionally, the close similarity (83% deduced amino-acid identity) but the pronounced difference in the expression patterns of these two fungal lectins during fruiting-body formation affords a suitable object for study of the relation of structural difference to the observed functional disparity in the same organism. Both galectins were crystallized after recombinant production. Crystals belong to either the orthorhombic space group C222(1) (Cgl-1) or the monoclinic space group P2(1) (Cgl-2). The latter crystals diffracted to 1.6 A resolution using synchrotron radiation. To solve the phasing problem, a selenomethionine-containing variant of Cgl-1 was designed. Crystals isomorphous to those of the native counterpart were obtained. Their structural analysis will also be crucial to solving the structure of Cgl-2.

  10. On Secret Sharing with Nonlinear Product Reconstruction

    DEFF Research Database (Denmark)

    Cascudo Pueyo, Ignacio; Cramer, Ronald; Mirandola, Diego

    2015-01-01

    Multiplicative linear secret sharing is a fundamental notion in the area of secure multiparty computation and, since recently, in the area of two-party cryptography as well. In a nutshell, this notion guarantees that the product of two secrets is obtained as a linear function of the vector consis...

  11. Primary endocrine-secreting pancreatic tumors.

    Science.gov (United States)

    Macaron, C

    1980-04-01

    Insulinoma, glucagonoma, gastrinoma (Zollinger-Ellison syndrome), vipoma, somatostatinoma and a tumor that secretes human pancreatic polypeptide are the primary endocrine-secreting tumors of the pancreas. hormones are produced by specific tumor cell types and cause a variety of dramatic clinical pictures. Diagnosis often requires hormone assays. Computerized tomography may be helpful. Definitive surgical treatment is possible, but metastases may be present.

  12. Family Secrets: The Bioethics of Genetic Testing

    Science.gov (United States)

    Markowitz, Dina G.; DuPre, Michael J.; Holt, Susan; Chen, Shaw-Ree; Wischnowski, Michael

    2006-01-01

    This article discusses "Family Secrets," a problem-based learning (PBL) curriculum module that focuses on the bioethical implications of genetic testing. In high school biology classrooms throughout New York State, students are using "Family Secrets" to learn about DNA testing; Huntington's disease (HD); and the ethical, legal,…

  13. The Secret in the Information Society

    NARCIS (Netherlands)

    D.W.J. Broeders (Dennis)

    2016-01-01

    textabstractWho can still keep a secret in a world in which everyone and everything are connected by technology aimed at charting and cross-referencing people, objects, movements, behaviour, relationships, tastes and preferences? The possibilities to keep a secret have come under severe pressure in

  14. Analysis of Secreted Proteins Using SILAC

    DEFF Research Database (Denmark)

    Henningsen, Jeanette; Blagoev, Blagoy; Kratchmarova, Irina

    2014-01-01

    Secreted proteins serve a crucial role in the communication between cells, tissues, and organs. Proteins released to the extracellular environment exert their function either locally or at distant points of the organism. Proteins are secreted in a highly dynamic fashion by cells and tissues...

  15. Characterization of a secreted Chlamydia protease

    DEFF Research Database (Denmark)

    Shaw, Allan C; Vandahl, Brian; Larsen, Martin Røssel

    2002-01-01

    Chlamydiae are obligate intracellular bacteria that are important human pathogens. The Chlamydia genomes contain orthologues to secretion apparatus proteins from other intracellular bacteria, but only a few secreted proteins have been identified. Most likely, effector proteins are secreted in order...... to promote infection. Effector proteins cannot be identified by motif or similarity searches. As a new strategy for identification of secreted proteins we have compared 2D-PAGE profiles of [35S]-labelled Chlamydia proteins from whole lysates of infected cells to 2D-PAGE profiles of proteins from purified...... Chlamydia. Several secretion candidates from Chlamydia trachomatis D and Chlamydia pneumoniae were detected by this method. Two protein spots were identified among the candidates. These represent fragments of the 'chlamydial protease- or proteasome-like activity factor' (CPAF) and were clearly present in 2D...

  16. Current Therapies That Modify Glucagon Secretion

    DEFF Research Database (Denmark)

    Grøndahl, Magnus F; Keating, Damien J; Vilsbøll, Tina

    2017-01-01

    and provide insights into how antidiabetic drugs influence glucagon secretion as well as a perspective on the future of glucagon-targeting drugs. RECENT FINDINGS: Several older as well as recent investigations have evaluated the effect of antidiabetic agents on glucagon secretion to understand how glucagon...... may be involved in the drugs' efficacy and safety profiles. Based on these findings, modulation of glucagon secretion seems to play a hitherto underestimated role in the efficacy and safety of several glucose-lowering drugs. Numerous drugs currently available to diabetologists are capable of altering...... glucagon secretion: metformin, sulfonylurea compounds, insulin, glucagon-like peptide-1 receptor agonists, dipeptidyl peptidase-4 inhibitors, sodium-glucose cotransporter 2 inhibitors and amylin mimetics. Their diverse effects on glucagon secretion are of importance for their individual efficacy and safety...

  17. Toxins and Secretion Systems of Photorhabdus luminescens

    Directory of Open Access Journals (Sweden)

    Athina Rodou

    2010-06-01

    Full Text Available Photorhabdus luminescens is a nematode-symbiotic, gram negative, bioluminescent bacterium, belonging to the family of Enterobacteriaceae.Recent studies show the importance of this bacterium as an alternative source of insecticides, as well as an emerging human pathogen. Various toxins have been identified and characterized in this bacterium. These toxins are classified into four major groups: the toxin complexes (Tcs, the Photorhabdus insect related (Pir proteins, the “makes caterpillars floppy” (Mcf toxins and the Photorhabdus virulence cassettes (PVC; the mechanisms however of toxin secretion are not fully elucidated. Using bioinformatics analysis and comparison against the components of known secretion systems, multiple copies of components of all known secretion systems, except the ones composing a type IV secretion system, were identified throughout the entire genome of the bacterium. This indicates that Photorhabdus luminescens has all the necessary means for the secretion of virulence factors, thus it is capable of establishing a microbial infection.

  18. Toxins and secretion systems of Photorhabdus luminescens.

    Science.gov (United States)

    Rodou, Athina; Ankrah, Dennis O; Stathopoulos, Christos

    2010-06-01

    Photorhabdus luminescens is a nematode-symbiotic, gram negative, bioluminescent bacterium, belonging to the family of Enterobacteriaceae. Recent studies show the importance of this bacterium as an alternative source of insecticides, as well as an emerging human pathogen. Various toxins have been identified and characterized in this bacterium. These toxins are classified into four major groups: the toxin complexes (Tcs), the Photorhabdus insect related (Pir) proteins, the "makes caterpillars floppy" (Mcf) toxins and the Photorhabdus virulence cassettes (PVC); the mechanisms however of toxin secretion are not fully elucidated. Using bioinformatics analysis and comparison against the components of known secretion systems, multiple copies of components of all known secretion systems, except the ones composing a type IV secretion system, were identified throughout the entire genome of the bacterium. This indicates that Photorhabdus luminescens has all the necessary means for the secretion of virulence factors, thus it is capable of establishing a microbial infection.

  19. Non-classical protein secretion in bacteria

    Directory of Open Access Journals (Sweden)

    Fausbøll Anders

    2005-10-01

    Full Text Available Abstract Background We present an overview of bacterial non-classical secretion and a prediction method for identification of proteins following signal peptide independent secretion pathways. We have compiled a list of proteins found extracellularly despite the absence of a signal peptide. Some of these proteins also have known roles in the cytoplasm, which means they could be so-called "moon-lightning" proteins having more than one function. Results A thorough literature search was conducted to compile a list of currently known bacterial non-classically secreted proteins. Pattern finding methods were applied to the sequences in order to identify putative signal sequences or motifs responsible for their secretion. We have found no signal or motif characteristic to any majority of the proteins in the compiled list of non-classically secreted proteins, and conclude that these proteins, indeed, seem to be secreted in a novel fashion. However, we also show that the apparently non-classically secreted proteins are still distinguished from cellular proteins by properties such as amino acid composition, secondary structure and disordered regions. Specifically, prediction of disorder reveals that bacterial secretory proteins are more structurally disordered than their cytoplasmic counterparts. Finally, artificial neural networks were used to construct protein feature based methods for identification of non-classically secreted proteins in both Gram-positive and Gram-negative bacteria. Conclusion We present a publicly available prediction method capable of discriminating between this group of proteins and other proteins, thus allowing for the identification of novel non-classically secreted proteins. We suggest candidates for non-classically secreted proteins in Escherichia coli and Bacillus subtilis. The prediction method is available online.

  20. Bile salts secretion in cirrhosis.

    Science.gov (United States)

    Correia, J P; Areias, E; Meneses, L; Tiago, E

    1977-02-01

    The bile salts secretion was studied in ten normal subjects and sixteen patients with alcoholic cirrhosis, in a basal period and during 60 minutes after Secretin injection. Total bile salts were measured by a modification of the enzymatic method of Iwata and Yamasaki and the individual bile salts were separated by silica gel thin-layer chromatography. During the 60 minutes after Secretin the mean concentration was 2.88 +/- 2.58 muM/ml in normals and 1.96 +/- 1.25 muM/ml in cirrhotics. The difference is not significant. During the first 20 minutes however the concentration was higher than 3 muM/ml in 8 out of 10 normals and lower than 2 muM/ml in 10 out 16 cirrhotics. The ratios of tri-to dihydroxy bile salts was similar in both groups. The ratios between bile salts conjugated with glycine and with taurine was higher in normals, and the ratio between free to conjugated bile salts was higher in cirrhotics. The lower concentration of total bile salts immediatly after Secretin, the higher proportion of taurin conjugates and of free bile salts could be important factors in the difficulties of fact digestion and absorption frequently found in patients with alcoholic cirrhosis.

  1. Identification of protein secretion systems and novel secreted proteins in Rhizobium leguminosarum bv. viciae

    Directory of Open Access Journals (Sweden)

    Krehenbrink Martin

    2008-01-01

    Full Text Available Abstract Background Proteins secreted by bacteria play an important role in infection of eukaryotic hosts. Rhizobia infect the roots of leguminous plants and establish a mutually beneficial symbiosis. Proteins secreted during the infection process by some rhizobial strains can influence infection and modify the plant defence signalling pathways. The aim of this study was to systematically analyse protein secretion in the recently sequenced strain Rhizobium leguminosarum bv. viciae 3841. Results Similarity searches using defined protein secretion systems from other Gram-negative bacteria as query sequences revealed that R. l. bv. viciae 3841 has ten putative protein secretion systems. These are the general export pathway (GEP, a twin-arginine translocase (TAT secretion system, four separate Type I systems, one putative Type IV system and three Type V autotransporters. Mutations in genes encoding each of these (except the GEP were generated, but only mutations affecting the PrsDE (Type I and TAT systems were observed to affect the growth phenotype and the profile of proteins in the culture supernatant. Bioinformatic analysis and mass fingerprinting of tryptic fragments of culture supernatant proteins identified 14 putative Type I substrates, 12 of which are secreted via the PrsDE, secretion system. The TAT mutant was defective for the symbiosis, forming nodules incapable of nitrogen fixation. Conclusion None of the R. l. bv. viciae 3841 protein secretion systems putatively involved in the secretion of proteins to the extracellular space (Type I, Type IV, Type V is required for establishing the symbiosis with legumes. The PrsDE (Type I system was shown to be the major route of protein secretion in non-symbiotic cells and to secrete proteins of widely varied size and predicted function. This is in contrast to many Type I systems from other bacteria, which typically secrete specific substrates encoded by genes often localised in close proximity to

  2. Identification of protein secretion systems and novel secreted proteins in Rhizobium leguminosarum bv. viciae.

    Science.gov (United States)

    Krehenbrink, Martin; Downie, J Allan

    2008-01-29

    Proteins secreted by bacteria play an important role in infection of eukaryotic hosts. Rhizobia infect the roots of leguminous plants and establish a mutually beneficial symbiosis. Proteins secreted during the infection process by some rhizobial strains can influence infection and modify the plant defence signalling pathways. The aim of this study was to systematically analyse protein secretion in the recently sequenced strain Rhizobium leguminosarum bv. viciae 3841. Similarity searches using defined protein secretion systems from other Gram-negative bacteria as query sequences revealed that R. l. bv. viciae 3841 has ten putative protein secretion systems. These are the general export pathway (GEP), a twin-arginine translocase (TAT) secretion system, four separate Type I systems, one putative Type IV system and three Type V autotransporters. Mutations in genes encoding each of these (except the GEP) were generated, but only mutations affecting the PrsDE (Type I) and TAT systems were observed to affect the growth phenotype and the profile of proteins in the culture supernatant. Bioinformatic analysis and mass fingerprinting of tryptic fragments of culture supernatant proteins identified 14 putative Type I substrates, 12 of which are secreted via the PrsDE, secretion system. The TAT mutant was defective for the symbiosis, forming nodules incapable of nitrogen fixation. None of the R. l. bv. viciae 3841 protein secretion systems putatively involved in the secretion of proteins to the extracellular space (Type I, Type IV, Type V) is required for establishing the symbiosis with legumes. The PrsDE (Type I) system was shown to be the major route of protein secretion in non-symbiotic cells and to secrete proteins of widely varied size and predicted function. This is in contrast to many Type I systems from other bacteria, which typically secrete specific substrates encoded by genes often localised in close proximity to the genes encoding the secretion system itself.

  3. THE BUFFER CAPACITY OF AIRWAY EPITHELIAL SECRETIONS

    Directory of Open Access Journals (Sweden)

    Dusik eKim

    2014-06-01

    Full Text Available The pH of airway epithelial secretions influences bacterial killing and mucus properties and is reduced by acidic pollutants, gastric reflux, and respiratory diseases such as cystic fibrosis (CF. The effect of acute acid loads depends on buffer capacity, however the buffering of airway secretions has not been well characterized. In this work we develop a method for titrating micro-scale (30 µl volumes and use it to study fluid secreted by the human airway epithelial cell line Calu-3, a widely used model for submucosal gland serous cells. Microtitration curves revealed that HCO3- is the major buffer. Peak buffer capacity (β increased from 17 to 28 mM/pH during forskolin stimulation, and was reduced by >50% in fluid secreted by cystic fibrosis transmembrane conductance regulator (CFTR-deficient Calu-3 monolayers, confirming an important role of CFTR in HCO3- secretion. Back-titration with NaOH revealed non-volatile buffer capacity due to proteins synthesized and released by the epithelial cells. Lysozyme and mucin concentrations were too low to buffer Calu-3 fluid significantly, however model titrations of porcine gastric mucins at concentrations near the sol-gel transition suggest that mucins may contribute to the buffer capacity of ASL in vivo. We conclude that CFTR-dependent HCO3- secretion and epithelially-derived proteins are the predominant buffers in Calu-3 secretions.

  4. Concentrations of ceftibuten in bronchial secretions.

    Science.gov (United States)

    Scaglione, F; Triscari, F; Demartini, G; Arcidiacono, M; Cocuzza, C; Fraschini, F

    1995-01-01

    Ceftibuten is a broad-spectrum oral cephalosporin exhibiting antimicrobial activity against a wide range of gram-negative and some gram-positive pathogens. Pharmacokinetic studies have shown that the molecule has an oral bioavailability higher than 90% of the administered dose (reaching peak serum concentrations of 5-19 mg/l after a single dose of 200 and 400 mg). Moreover, ceftibuten has been shown to be useful in the treatment of acute lower respiratory tract infections. This study was performed to determine the distribution of ceftibuten in bronchial secretions from patients affected by the exacerbation of chronic bronchitis. Patients were treated with a single 400-mg oral dose of ceftibuten. Blood and bronchial-secretion samples were obtained just before, and at 0.5, 1, 2, 4, 8, 12, 16 and 24 h after dosing. Cells were separated from bronchial secretions by centrifugation. Ceftibuten in duplicate samples of both serum and bronchial secretion was quantified by HPLC. Ceftibuten reached peak levels 2 and 4 h after oral administration in serum and in bronchial secretions, respectively (18.12 +/- 2.13 and 9.19 +/- 3.1 mg/l, respectively). Falling curves after the peaks showed a monoexponential decay. The absorption was very rapid both in serum and bronchial secretions, but elimination was slower in bronchial secretions than in serum.

  5. The buffer capacity of airway epithelial secretions.

    Science.gov (United States)

    Kim, Dusik; Liao, Jie; Hanrahan, John W

    2014-01-01

    The pH of airway epithelial secretions influences bacterial killing and mucus properties and is reduced by acidic pollutants, gastric reflux, and respiratory diseases such as cystic fibrosis (CF). The effect of acute acid loads depends on buffer capacity, however the buffering of airway secretions has not been well characterized. In this work we develop a method for titrating micro-scale (30 μl) volumes and use it to study fluid secreted by the human airway epithelial cell line Calu-3, a widely used model for submucosal gland serous cells. Microtitration curves revealed that HCO(-) 3 is the major buffer. Peak buffer capacity (β) increased from 17 to 28 mM/pH during forskolin stimulation, and was reduced by >50% in fluid secreted by cystic fibrosis transmembrane conductance regulator (CFTR)-deficient Calu-3 monolayers, confirming an important role of CFTR in HCO(-) 3 secretion. Back-titration with NaOH revealed non-volatile buffer capacity due to proteins synthesized and released by the epithelial cells. Lysozyme and mucin concentrations were too low to buffer Calu-3 fluid significantly, however model titrations of porcine gastric mucins at concentrations near the sol-gel transition suggest that mucins may contribute to the buffer capacity of ASL in vivo. We conclude that CFTR-dependent HCO(-) 3 secretion and epithelially-derived proteins are the predominant buffers in Calu-3 secretions.

  6. Lycaenid Caterpillar Secretions Manipulate Attendant Ant Behavior.

    Science.gov (United States)

    Hojo, Masaru K; Pierce, Naomi E; Tsuji, Kazuki

    2015-08-31

    Mutualistic interactions typically involve the exchange of different commodities between species. Nutritious secretions are produced by a number of insects and plants in exchange for services such as defense. These rewards are valuable metabolically and can be used to reinforce the behavior of symbiotic partners that can learn and remember them effectively. We show here novel effects of insect exocrine secretions produced by caterpillars in modulating the behavior of attendant ants in the food-for-defense interaction between lycaenid butterflies and ants. Reward secretions from the dorsal nectary organ (DNO) of Narathura japonica caterpillars function to reduce the locomotory activities of their attendant ants, Pristomyrmex punctatus workers. Moreover, workers that feed from caterpillar secretions are significantly more likely to show aggressive responses to eversion of the tentacle organs of the caterpillars. Analysis of the neurogenic amines in the brains of workers that consumed caterpillar secretions showed a significant decrease in levels of dopamine compared with controls. Experimental treatments in which reserpine, a known inhibitor of dopamine in Drosophila, was fed to workers similarly reduced their locomotory activity. We conclude that DNO secretions of lycaenid caterpillars can manipulate attendant ant behavior by altering dopaminergic regulation and increasing partner fidelity. Unless manipulated ants also receive a net nutritional benefit from DNO secretions, this suggests that similar reward-for-defense interactions that have been traditionally considered to be mutualisms may in fact be parasitic in nature. Copyright © 2015 Elsevier Ltd. All rights reserved.

  7. [Mechanism of alcohol action on gastric secretion].

    Science.gov (United States)

    Vasilevskaia, L S; Skurikhin, I M; Guliev, R R

    2001-01-01

    In chronic experiment on dogs (3 dogs with Pavlov's miniature stomach, 3 dogs with Heidenhain's miniature stomach) the mechanism of action of alcohol (8%-150 ml) on gastric secretion was clarified. For this purpose the new inhibitor of gastric secretion--glycopeptide was utilized, which action was preset in laboratory of G.K. Shlygin. Was shown, that in effect of alcohol on gastric secretion take place the complicated mechanism including as a nervous regulation (vagus nerves), and humoral: participation of gastrin and histamine in secretory effect of a stomach, and also immediate effect of ethanol on acid glands.

  8. Regulation of glucagon secretion by incretins

    DEFF Research Database (Denmark)

    Holst, Jens Juul; Christensen, M; Lund, A

    2011-01-01

    Glucagon secretion plays an essential role in the regulation of hepatic glucose production, and elevated fasting and postprandial plasma glucagon concentrations in patients with type 2 diabetes (T2DM) contribute to their hyperglycaemia. The reason for the hyperglucagonaemia is unclear, but recent...... that endogenous GLP-1 plays an important role in regulation of glucagon secretion during fasting as well as postprandially. The mechanisms whereby GLP-1 regulates glucagon secretion are debated, but studies in isolated perfused rat pancreas point to an important role for a paracrine regulation by somatostatin...

  9. Characterization of the Wild-Type and Truncated Forms of a Neutral GH10 Xylanase from Coprinus cinereus: Roles of C-Terminal Basic Amino Acid-Rich Extension in Its SDS Resistance, Thermostability, and Activity.

    Science.gov (United States)

    Hu, Hang; Chen, Kaixiang; Li, Lulu; Long, Liangkun; Ding, Shaojun

    2017-04-28

    A neutral xylanase (CcXyn) was identified from Coprinus cinereus. It has a single GH10 catalytic domain with a basic amino acid-rich extension (PVRRK) at the C-terminus. In this study, the wild-type (CcXyn) and C-terminus-truncated xylanase (CcXyn-Δ5C) were heterologously expressed in Pichia pastoris and their characteristics were comparatively analyzed with aims to examine the effect of this extension on the enzyme function. The circular dichorism analysis indicated that both enzymes in general had a similar structure, but CcXyn-Δ5C contained less α-helices (42.9%) and more random coil contents (35.5%) than CcXyn (47.0% and 32.8%, respectively). Both enzymes had the same pH (7.0) and temperature (45°C) optima, and similar substrate specificity on different xylans. They all hydrolyzed beechwood xylan primarily to xylobiose and xylotriose. The amounts of xylobiose and xylotriose accounted for 91.5% and 92.2% (w/w) of total xylooligosaccharides (XOS) generated from beechwood by CcXyn and CcXyn-Δ5C, respectively. However, truncation of the C-terminal 5-amino-acids extension significantly improved the thermostability, SDS resistance, and pH stability at pH 6.0-9.0. Furthermore, CcXyn-Δ5C exhibited a much lower Km value than CcXyn (0.27 mg/ml vs 0.83 mg/ml), and therefore, the catalytic efficiency of CcXyn-Δ5C was 2.4-times higher than that of CcXyn. These properties make CcXyn-Δ5C a good model for the structure-function study of (α/β)8-barrel-folded enzymes and a promising candidate for various applications, especially in the detergent industry and XOS production.

  10. Paramagnetic 13C and 15N NMR analyses of the push and pull effects in cytochrome c peroxidase and Coprinus cinereus peroxidase variants: functional roles of highly conserved amino acids around heme.

    Science.gov (United States)

    Nonaka, Daisuke; Wariishi, Hiroyuki; Welinder, Karen G; Fujii, Hiroshi

    2010-01-12

    Paramagnetic (13)C and (15)N nuclear magnetic resonance (NMR) spectroscopy of heme-bound cyanide ((13)C(15)N) was applied to 11 cytochrome c peroxidase (CcP) and Coprinus cinereus peroxidase (CIP) mutants to investigate contributions to the push and pull effects of conserved amino acids around heme. The (13)C and (15)N NMR data for the distal His and Arg mutants indicated that distal His is the key amino acid residue creating the strong pull effect and that distal Arg assists. The mutation of distal Trp of CcP to Phe, the amino acid at this position in CIP, changed the push and pull effects so they resembled those of CIP, whereas the mutation of distal Phe of CIP to Trp changed this mutant to become CcP-like. The (13)C NMR shifts for the proximal Asp mutants clearly showed that the proximal Asp-His hydrogen bonding strengthens the push effect. However, even in the absence of a hydrogen bond, the push effect of proximal His in peroxidase is significantly stronger than in globins. Comparison of these NMR data with the compound I formation rate constants and crystal structures of these mutants showed that (1) the base catalysis of the distal His is more critical for rapid compound I formation than its acid catalysis, (2) the primary function of the distal Arg is to maintain the distal heme pocket in favor of rapid compound I formation via hydrogen bonding, and (3) the push effect is the major contributor to the differential rates of compound I formation in wild-type peroxidases.

  11. Lycaenid Caterpillar Secretions Manipulate Attendant Ant Behavior

    National Research Council Canada - National Science Library

    Hojo, Masaru K; Pierce, Naomi E; Tsuji, Kazuki

    2015-01-01

    .... We show here novel effects of insect exocrine secretions produced by caterpillars in modulating the behavior of attendant ants in the food-for-defense interaction between lycaenid butterflies and ants...

  12. Acetylcholine regulates ghrelin secretion in humans

    NARCIS (Netherlands)

    F. Broglio (Fabio); E. Ghigo (Ezio); C. Gottero; F. Prodam (Flavia); S. Destefanis; A. Benso; C. Gauna (Carlotta); L.J. Hofland (Leo); E. Arvat; A-J. van der Lely (Aart-Jan); P.M. van Koetsveld (Peter)

    2004-01-01

    textabstractGhrelin secretion has been reportedly increased by fasting and energy restriction but decreased by food intake, glucose, insulin, and somatostatin. However, its regulation is still far from clarified. The cholinergic system mediates some ghrelin actions, e.g.

  13. Cell Secretion: Current Structural and Biochemical Insights

    Directory of Open Access Journals (Sweden)

    Saurabh Trikha

    2010-01-01

    Full Text Available Essential physiological functions in eukaryotic cells, such as release of hormones and digestive enzymes, neurotransmission, and intercellular signaling, are all achieved by cell secretion. In regulated (calcium-dependent secretion, membrane-bound secretory vesicles dock and transiently fuse with specialized, permanent, plasma membrane structures, called porosomes or fusion pores. Porosomes are supramolecular, cup-shaped lipoprotein structures at the cell plasma membrane that mediate and control the release of vesicle cargo to the outside of the cell. The sizes of porosomes range from 150nm in diameter in acinar cells of the exocrine pancreas to 12nm in neurons. In recent years, significant progress has been made in our understanding of the porosome and the cellular activities required for cell secretion, such as membrane fusion and swelling of secretory vesicles. The discovery of the porosome complex and the molecular mechanism of cell secretion are summarized in this article.

  14. Microbial cell surfaces and secretion systems

    NARCIS (Netherlands)

    Tommassen, J.P.M.|info:eu-repo/dai/nl/069127077; Wosten, H.A.B.|info:eu-repo/dai/nl/120693186

    2015-01-01

    Microbial cell surfaces, surface-exposed organelles, and secreted proteins are important for the interaction with the environment, including adhesion to hosts, protection against host defense mechanisms, nutrient acquisition, and intermicrobial competition. Here, we describe the structures of the

  15. Protein Secretion and the Endoplasmic Reticulum

    OpenAIRE

    Benham, Adam M.

    2012-01-01

    In a complex multicellular organism, different cell types engage in specialist functions, and as a result, the secretory output of cells and tissues varies widely. Whereas some quiescent cell types secrete minor amounts of proteins, tissues like the pancreas, producing insulin and other hormones, and mature B cells, producing antibodies, place a great demand on their endoplasmic reticulum (ER). Our understanding of how protein secretion in general is controlled in the ER is now quite sophisti...

  16. Transepithelial Bicarbonate Secretion: Lessons from the Pancreas

    Science.gov (United States)

    Park, Hyun Woo; Lee, Min Goo

    2012-01-01

    Many cystic fibrosis transmembrane conductance regulator (CFTR)-expressing epithelia secrete bicarbonate (HCO3−)-containing fluids. Recent evidence suggests that defects in epithelial bicarbonate secretion are directly involved in the pathogenesis of cystic fibrosis, in particular by building up hyperviscous mucus in the ductal structures of the lung and pancreas. Pancreatic juice is one of the representative fluids that contain a very high concentration of bicarbonate among bodily fluids that are secreted from CFTR-expressing epithelia. We introduce up-to-date knowledge on the basic principles of transepithelial bicarbonate transport by showing the mechanisms involved in pancreatic bicarbonate secretion. The model of pancreatic bicarbonate secretion described herein may also apply to other exocrine epithelia. As a central regulator of bicarbonate transport at the apical membrane, CFTR plays an essential role in both direct and indirect bicarbonate secretion. The major role of CFTR in bicarbonate secretion would be variable depending on the tissue and cell type. For example, in epithelial cells that produce a low concentration of bicarbonate-containing fluid (up to 80 mm), either CFTR-dependent Cl−/HCO3− exchange or CFTR anion channel with low bicarbonate permeability would be sufficient to generate such fluid. However, in cells that secrete high-bicarbonate-containing fluids, a highly selective CFTR bicarbonate channel activity is required. Therefore, understanding the molecular mechanism of transepithelial bicarbonate transport and the role of CFTR in each specific epithelium will provide therapeutic strategies to recover from epithelial defects induced by hyposecretion of bicarbonate in cystic fibrosis. PMID:23028131

  17. Peptides and neurotransmitters that affect renin secretion

    Science.gov (United States)

    Ganong, W. F.; Porter, J. P.; Bahnson, T. D.; Said, S. I.

    1984-01-01

    Substance P inhibits renin secretion. This polypeptide is a transmitter in primary afferent neurons and is released from the peripheral as well as the central portions of these neurons. It is present in afferent nerves from the kidneys. Neuropeptide Y, which is a cotransmitter with norepinephrine and epinephrine, is found in sympathetic neurons that are closely associated with and presumably innervate the juxtagolmerular cells. Its effect on renin secretion is unknown, but it produces renal vasoconstriction and natriuresis. Vasoactive intestinal polypeptide (VIP) is a cotransmitter with acetylocholine in cholinergic neurons, and this polypeptide stimulates renin secretion. We cannot find any evidence for its occurence in neurons in the kidneys, but various stimuli increase plasma VIP to levels comparable to those produced by doses of exogenous VIP which stimulated renin secretion. Neostigmine increases plasma VIP and plasma renin activity, and the VIP appears to be responsible for the increase in renin secretion, since the increase is not blocked by renal denervation or propranolol. Stimulation of various areas in the brain produces sympathetically mediated increases in plasma renin activity associated with increases in blood pressure. However, there is pharmacological evidence that the renin response can be separated from the blood pressure response. In anaesthetized dogs, drugs that increase central serotonergic discharge increase renin secretion without increasing blood pressure. In rats, activation of sertonergic neurons in the dorsal raphe nucleus increases renin secretion by a pathway that projects from this nucleus to the ventral hypothalamus, and from there to the kidneys via the sympathetic nervous system. The serotonin releasing drug parachloramphetamine also increases plasma VIP, but VIP does not appear to be the primary mediator of the renin response. There is preliminary evidence that the serotonergic neurons in the dorsal raphe nucleus are part of the

  18. Cholecystokinin inhibits gastrin secretion independently of paracrine somatostatin secretion in the pig

    DEFF Research Database (Denmark)

    Schmidt, P T; Hansen, L; Hilsted, L

    2004-01-01

    BACKGROUND: Cholecystokinin inhibits the secretion of gastrin from antral G cells, an effect that is speculated to be mediated by D cells secreting somatostatin. The aim of the study was to test directly whether cholecystokinin inhibition of antral gastrin secretion is mediated by somatostatin....... METHODS: The effects of CCK on gastrin and somatostatin secretion were studied in isolated vascularly perfused preparations of pig antrum before and after immunoneutralization brought about by infusion of large amounts of a high affinity monoclonal antibody against somatostatin. RESULTS: CCK infusion...... at 10(-9) M and 10(-8) M decreased gastrin output to 70.5% +/- 7.6% (n = 8) and 76.3% +/- 3.6% (n = 7) of basal output, respectively. CCK at 10(-10) M had no effect (n = 6). Somatostatin secretion was dose-dependently increased by CCK infusion and increased to 268 +/- 38.2% (n = 7) of basal secretion...

  19. Development of secreted proteins as biotherapeutic agents.

    Science.gov (United States)

    Bonin-Debs, Angelika L; Boche, Irene; Gille, Hendrik; Brinkmann, Ulrich

    2004-04-01

    As one of the most important classes of proteins, secreted factors account for about one-tenth of the human genome, 3000 - 4000 in total, including factors of signalling pathways, blood coagulation and immune defence, as well as digestive enzymes and components of the extracellular matrix. Secreted proteins are a rich source of new therapeutics and drug targets, and are currently the focus of major drug discovery programmes throughout the industry. Many of the most important novel drugs developed in biotechnology have resulted from the application of secreted proteins as therapeutics. Secreted proteins often circulate throughout the body and, therefore, have access to most organs and tissues. Because of that, many of the factors are themselves therapeutic agents. This paper gives an overview on the features and functions of human secreted proteins and peptides, as well as strategies by which to discover additional therapeutic proteins from the human 'secretome'. Furthermore, a variety of examples are provided for the therapeutic use of recombinant secreted proteins as 'biologicals', including features and applications of recombinant antibodies, erythropoietin, insulin, interferon, plasminogen activators, growth hormone and colony-stimulating factors.

  20. Substrate adaptation of Trichophyton rubrum secreted endoproteases.

    Science.gov (United States)

    Chen, Jian; Yi, Jinling; Liu, Li; Yin, Songchao; Chen, Rongzhang; Li, Meirong; Ye, Congxiu; Zhang, Yu-qing; Lai, Wei

    2010-02-01

    Trichophyton rubrum is the most common pathogen caused the dermatophytosis of nail and skin in human. The secreted proteases were considered to be the most important virulence factors. However, the substrates adaptation of T. rubrum secreted proteases is largely unknown. For the first time, we use the keratins from human nail and skin stratum corneum as the growth medium to investigate the different expression patterns of T. rubrum secreted endoproteases genes. During grow in both keratin-containing media SUB7 and MEP2 were the highest expressed gene in each family. These results indicated that SUB7 and MEP2 may be the dominant endoproteases secreted by T. rubrum during host infection and the other proteases may play a supplementary role. The direct comparison of T. rubrum grown on skin and nail medium showed different substrate favorite of secreted endoproteases. The genes MEP2, SUB5, SUB2 and SUB3 were more active during growth in skin medium, possibly these proteases have a higher affinity for skin original keratins. While the structures of SUB1, SUB4, and MEP4 maybe more suitable for the degradation of nail original keratins. This work presents useful molecular details for further understanding the pathogenesis of secreted proteases and the wide adaptation of T. rubrum.

  1. Type VI Secretion System in Pseudomonas aeruginosa

    Science.gov (United States)

    Hachani, Abderrahman; Lossi, Nadine S.; Hamilton, Alexander; Jones, Cerith; Bleves, Sophie; Albesa-Jové, David; Filloux, Alain

    2011-01-01

    Pseudomonas aeruginosa is a Gram-negative bacterium causing chronic infections in cystic fibrosis patients. Such infections are associated with an active type VI secretion system (T6SS), which consists of about 15 conserved components, including the AAA+ ATPase, ClpV. The T6SS secretes two categories of proteins, VgrG and Hcp. Hcp is structurally similar to a phage tail tube component, whereas VgrG proteins show similarity to the puncturing device at the tip of the phage tube. In P. aeruginosa, three T6SSs are known. The expression of H1-T6SS genes is controlled by the RetS sensor. Here, 10 vgrG genes were identified in the PAO1 genome, among which three are co-regulated with H1-T6SS, namely vgrG1a/b/c. Whereas VgrG1a and VgrG1c were secreted in a ClpV1-dependent manner, secretion of VgrG1b was ClpV1-independent. We show that VgrG1a and VgrG1c form multimers, which confirmed the VgrG model predicting trimers similar to the tail spike. We demonstrate that Hcp1 secretion requires either VgrG1a or VgrG1c, which may act independently to puncture the bacterial envelope and give Hcp1 access to the surface. VgrG1b is not required for Hcp1 secretion. Thus, VgrG1b does not require H1-T6SS for secretion nor does H1-T6SS require VgrG1b for its function. Finally, we show that VgrG proteins are required for secretion of a genuine H1-T6SS substrate, Tse3. Our results demonstrate that VgrG proteins are not only secreted components but are essential for secretion of other T6SS substrates. Overall, we emphasize variability in behavior of three P. aeruginosa VgrGs, suggesting that, although very similar, distinct VgrGs achieve specific functions. PMID:21325275

  2. Exocytosis and protein secretion in Trypanosoma

    Directory of Open Access Journals (Sweden)

    Rossignol Michel

    2010-01-01

    Full Text Available Abstract Background Human African trypanosomiasis is a lethal disease caused by the extracellular parasite Trypanosoma brucei. The proteins secreted by T. brucei inhibit the maturation of dendritic cells and their ability to induce lymphocytic allogenic responses. To better understand the pathogenic process, we combined different approaches to characterize these secreted proteins. Results Overall, 444 proteins were identified using mass spectrometry, the largest parasite secretome described to date. Functional analysis of these proteins revealed a strong bias toward folding and degradation processes and to a lesser extent toward nucleotide metabolism. These features were shared by different strains of T. brucei, but distinguished the secretome from published T. brucei whole proteome or glycosome. In addition, several proteins had not been previously described in Trypanosoma and some constitute novel potential therapeutic targets or diagnostic markers. Interestingly, a high proportion of these secreted proteins are known to have alternative roles once secreted. Furthermore, bioinformatic analysis showed that a significant proportion of proteins in the secretome lack transit peptide and are probably not secreted through the classical sorting pathway. Membrane vesicles from secretion buffer and infested rat serum were purified on sucrose gradient and electron microscopy pictures have shown 50- to 100-nm vesicles budding from the coated plasma membrane. Mass spectrometry confirmed the presence of Trypanosoma proteins in these microvesicles, showing that an active exocytosis might occur beyond the flagellar pocket. Conclusions This study brings out several unexpected features of the secreted proteins and opens novel perspectives concerning the survival strategy of Trypanosoma as well as possible ways to control the disease. In addition, concordant lines of evidence support the original hypothesis of the involvement of microvesicle-like bodies in the

  3. Sagnac secret sharing over telecom fiber networks.

    Science.gov (United States)

    Bogdanski, Jan; Ahrens, Johan; Bourennane, Mohamed

    2009-01-19

    We report the first Sagnac quantum secret sharing (in three-and four-party implementations) over 1550 nm single mode fiber (SMF) networks, using a single qubit protocol with phase encoding. Our secret sharing experiment has been based on a single qubit protocol, which has opened the door to practical secret sharing implementation over fiber telecom channels and in free-space. The previous quantum secret sharing proposals were based on multiparticle entangled states, difficult in the practical implementation and not scalable. Our experimental data in the three-party implementation show stable (in regards to birefringence drift) quantum secret sharing transmissions at the total Sagnac transmission loop distances of 55-75 km with the quantum bit error rates (QBER) of 2.3-2.4% for the mean photon number micro?= 0.1 and 1.7-2.1% for micro= 0.3. In the four-party case we have achieved quantum secret sharing transmissions at the total Sagnac transmission loop distances of 45-55 km with the quantum bit error rates (QBER) of 3.0-3.7% for the mean photon number micro= 0.1 and 1.8-3.0% for micro?= 0.3. The stability of quantum transmission has been achieved thanks to our new concept for compensation of SMF birefringence effects in Sagnac, based on a polarization control system and a polarization insensitive phase modulator. The measurement results have showed feasibility of quantum secret sharing over telecom fiber networks in Sagnac configuration, using standard fiber telecom components.

  4. Effects of endothelin family on ANP secretion.

    Science.gov (United States)

    Yuan, Kuichang; Park, Byung Mun; Choi, Young Tae; Kim, Jong Hun; Cho, Kyung Woo; Kim, Suhn Hee

    2016-08-01

    The endothelins (ET) peptide family consists of ET-1, ET-2, ET-3, and sarafotoxin (s6C, a snake venom) and their actions appears to be different among isoforms. The aim of this study was to compare the secretagogue effect of ET-1 on atrial natriuretic peptide (ANP) secretion with ET-3 and evaluate its physiological meaning. Isolated nonbeating atria from male Sprague-Dawley rats were used to evaluate stretch-activated ANP secretion in response to ET-1, ET-2, ET-3, and s6C. Changes in mean blood pressure (MAP) were measured during acute injection of ET-1 and ET-3 with and without natriuretic peptide receptor-A antagonist (A71915) in anesthetized rats. Changes in atrial volume induced by increased atrial pressure from o to 1, 2, 4, or 6cm H2O caused proportional increases in mechanically-stimulated extracellular fluid (ECF) translocation and stretch-activated ANP secretion. ET-1 (10nM) augmented basal and stretch-activated ANP secretion in terms of ECF translocation, which was blocked by the pretreatment with ETA receptor antagonist (BQ123, 1μM) but not by ETB receptor antagonist (BQ788, 1μM). ETA receptor antagonist itself suppressed stretch-activated ANP secretion. As compared to ET-1- induced ANP secretion (3.2-fold by 10nM), the secretagogue effects of ANP secretion by ET-2 was similar (2.8-fold by 10nM) and ET-3 and s6C were less potent (1.7-fold and 1.5-fold by 100nM, respectively). Acute injection of ET-1 or ET-3 increased mean blood pressure (MAP), which was augmented in the presence of natriuretic peptide receptor-A antagonist. Therefore, we suggest that the order of secretagogue effect of ET family on ANP secretion was ET-1≥ET-2>ET-3>s6C and ET-1-induced ANP secretion negatively regulates the pressor effect of ET-1. Copyright © 2016 Elsevier Inc. All rights reserved.

  5. Type VI Secretion System in Pseudomonas aeruginosa SECRETION AND MULTIMERIZATION OF VgrG PROTEINS

    NARCIS (Netherlands)

    Hachani, Abderrahman; Lossi, Nadine S.; Hamilton, Alexander; Jones, Cerith; Bleves, Sophie; Albesa-Jove, David; Filloux, Alain

    2011-01-01

    Pseudomonas aeruginosa is a Gram-negative bacterium causing chronic infections in cystic fibrosis patients. Such infections are associated with an active type VI secretion system (T6SS), which consists of about 15 conserved components, including the AAA(+) ATPase, ClpV. The T6SS secretes two

  6. Meaningful Share Generation for Increased Number of Secrets in Visual Secret-Sharing Scheme

    Directory of Open Access Journals (Sweden)

    Mustafa Ulutas

    2010-01-01

    Full Text Available This paper presents a new scheme for hiding two halftone secret images into two meaningful shares created from halftone cover images. Meaningful shares are more desirable than noise-like (meaningless shares in Visual Secret Sharing because they look natural and do not attract eavesdroppers' attention. Previous works in the field focus on either increasing number of secrets or creating meaningful shares for one secret image. The method outlined in this paper both increases the number of secrets and creates meaningful shares at the same time. While the contrast ratio of shares is equal to that of Extended Visual Cryptography, two secrets are encoded into two shares as opposed to one secret in the Extended Visual Cryptography. Any two natural-looking images can be used as cover unlike the Halftone Visual Cryptography method where one cover should be the negative of the other cover image and can only encode one secret. Effectiveness of the proposed method is verified by an experiment.

  7. Shared Secrets versus Secrets Kept Private Are Linked to Better Adolescent Adjustment

    Science.gov (United States)

    Frijns, Tom; Finkenauer, Catrin; Keijsers, Loes

    2013-01-01

    It is a household notion that secrecy is bad while sharing is good. But what about shared secrets? The present research adopts a functional analysis of sharing secrets, arguing that it should negate harmful consequences generally associated with secrecy and serves important interpersonal functions in adolescence. A survey study among 790 Dutch…

  8. Characterization of Nops, nodulation outer proteins, secreted via the type III secretion system of NGR234.

    Science.gov (United States)

    Marie, Corinne; Deakin, William J; Viprey, Virginie; Kopciñska, Joanna; Golinowski, Wladyslaw; Krishnan, Hari B; Perret, Xavier; Broughton, William J

    2003-09-01

    The nitrogen-fixing symbiotic bacterium Rhizobium species NGR234 secretes, via a type III secretion system (TTSS), proteins called Nops (nodulation outer proteins). Abolition of TTSS-dependent protein secretion has either no effect or leads to a change in the number of nodules on selected plants. More dramatically, Nops impair nodule development on Crotalaria juncea roots, resulting in the formation of nonfixing pseudonodules. A double mutation of nopX and nopL, which code for two previously identified secreted proteins, leads to a phenotype on Pachyrhizus tuberosus differing from that of a mutant in which the TTSS is not functional. Use of antibodies and a modification of the purification protocol revealed that NGR234 secretes additional proteins in a TTSS-dependent manner. One of them was identified as NopA, a small 7-kDa protein. Single mutations in nopX and nopL were also generated to assess the involvement of each Nop in protein secretion and nodule formation. Mutation of nopX had little effect on NopL and NopA secretion but greatly affected the interaction of NGR234 with many plant hosts tested. NopL was not necessary for the secretion of any Nops but was required for efficient nodulation of some plant species. NopL may thus act as an effector protein whose recognition is dependent upon the hosts' genetic background.

  9. Functional anatomy and physiology of gastric secretion.

    Science.gov (United States)

    Schubert, Mitchell L

    2015-11-01

    This review summarizes the past year's literature regarding the neuroendocrine and intracellular regulation of gastric acid secretion, discussing both basic and clinical aspects. Gastric acid facilitates the digestion of protein as well as the absorption of iron, calcium, vitamin B12, and certain medications. High acidity kills ingested microorganisms and limits bacterial overgrowth, enteric infection, and possibly spontaneous bacterial peritonitis. The main stimulants of acid secretion are gastrin, released from antral gastrin cells; histamine, released from oxyntic enterochromaffin-like cells; and acetylcholine, released from antral and oxyntic intramural neurons. Ghrelin and coffee also stimulate acid secretion whereas somatostatin, cholecystokinin, glucagon-like peptide-1, and atrial natriuretic peptide inhibit acid secretion. Although 95% of parietal cells are contained within the oxyntic mucosa (fundus and body), 50% of human antral glands contain parietal cells. Proton pump inhibitors are considered well tolerated drugs, but concerns have been raised regarding dysbiosis, atrophic gastritis, hypergastrinemia, hypomagnesemia, and enteritis/colitis. Our understanding of the functional anatomy and physiology of gastric secretion continues to advance. Such knowledge is crucial for improved management of acid-peptic disorders, prevention and management of neoplasia, and the development of novel medications.

  10. Fast-weighted secret image sharing

    Science.gov (United States)

    Lin, Sian-Jheng; Chen, Lee Shu-Teng; Lin, Ja-Chen

    2009-07-01

    Thien and Lin [Comput. and Graphics 26(5), 765-770 (2002)] proposed a threshold scheme to share a secret image among n shadows: any t of the n shadows can recover the secret, whereas t-1 or fewer shadows cannot. However, in real life, certain managers probably play key roles to run a company and thus need special authority to recover the secret in managers' meeting. (Each manager's shadow should be more powerful than an ordinary employee's shadow.) In Thien and Lin's scheme, if a company has less than t managers, then manager's meeting cannot recover the secret, unless some managers were given multiple shadows in advance. But this compromise causes managers inconvenience because too many shadows were to be kept daily and carried to the meeting. To solve this dilemma, a weighted sharing method is proposed: each of the shadows has a weight. The secret is recovered if and only if the total weights (rather than the number) of received shadows is at least t. The properties of GF(2r) are utilized to accelerate sharing speed. Besides, the method is also a more general approach to polynomial-based sharing. Moreover, for convenience, each person keeps only one shadow and only one shadow index.

  11. Melatonin Secretion Pattern in Critically Ill Patients

    DEFF Research Database (Denmark)

    Boyko, Yuliya; Holst, René; Jennum, Poul

    2017-01-01

    Critically ill patients have abnormal circadian and sleep homeostasis. This may be associated with higher morbidity and mortality. The aims of this pilot study were (1) to describe melatonin secretion in conscious critically ill mechanically ventilated patients and (2) to describe whether melaton...... secretion in these patients. Remifentanil did not affect melatonin secretion but was associated with lower risk of atypical sleep pattern. REM sleep was only registered during the period of nonsedation.......Critically ill patients have abnormal circadian and sleep homeostasis. This may be associated with higher morbidity and mortality. The aims of this pilot study were (1) to describe melatonin secretion in conscious critically ill mechanically ventilated patients and (2) to describe whether melatonin...... secretion and sleep patterns differed in these patients with and without remifentanil infusion. Eight patients were included. Blood-melatonin was taken every 4th hour, and polysomnography was carried out continually during a 48-hour period. American Academy of Sleep Medicine criteria were used for sleep...

  12. Melatonin Secretion Pattern in Critically Ill Patients

    DEFF Research Database (Denmark)

    Boyko, Yuliya; Holst, René; Jennum, Poul

    2017-01-01

    Critically ill patients have abnormal circadian and sleep homeostasis. This may be associated with higher morbidity and mortality. The aims of this pilot study were (1) to describe melatonin secretion in conscious critically ill mechanically ventilated patients and (2) to describe whether melatonin...... secretion and sleep patterns differed in these patients with and without remifentanil infusion. Eight patients were included. Blood-melatonin was taken every 4th hour, and polysomnography was carried out continually during a 48-hour period. American Academy of Sleep Medicine criteria were used for sleep...... scoring if sleep patterns were identified; otherwise, Watson's classification was applied. As remifentanil was periodically administered during the study, its effect on melatonin and sleep was assessed. Melatonin secretion in these patients followed a phase-delayed diurnal curve. We did not observe any...

  13. Standpoints and protection of business secrets

    Directory of Open Access Journals (Sweden)

    Brane Bertoncelj

    2001-06-01

    Full Text Available The human impact on an information system where data bases, containing business secretes, are stored is one of the most unreliable and unforeseeable factors. For this reason, it must not be underestimated. The results of this study indicate a correlation between behavioural intention and protection of business secretes. There is a statistically significant correlation between behavioural intention and behavioural supervision. This means that an increased level of perceived supervision over one's own behaviour is related to behavioural intention. A great majority of participants would not divulge a business secret due to internal moral factors, i.e., they possess the appropriate capabilities to determine the advantages of social moral values over personal values.

  14. Targeted Secretion Inhibitors—Innovative Protein Therapeutics

    Directory of Open Access Journals (Sweden)

    Foster Keith

    2010-12-01

    Full Text Available Botulinum neurotoxins are highly effective therapeutic products. Their therapeutic success results from highly specific and potent inhibition of neurotransmitter release with a duration of action measured in months. These same properties, however, make the botulinum neurotoxins the most potent acute lethal toxins known. Their toxicity and restricted target cell activity severely limits their clinical utility. Understanding the structure-function relationship of the neurotoxins has enabled the development of recombinant proteins selectively incorporating specific aspects of their pharmacology. The resulting proteins are not neurotoxins, but a new class of biopharmaceuticals, Targeted Secretion Inhibitors (TSI, suitable for the treatment of a wide range of diseases where secretion plays a major role. TSI proteins inhibit secretion for a prolonged period following a single application, making them particularly suited to the treatment of chronic diseases. A TSI for the treatment of chronic pain is in clinical development.

  15. Hiding secret data into a carrier image

    Directory of Open Access Journals (Sweden)

    Ovidiu COSMA

    2012-06-01

    Full Text Available The object of steganography is embedding hidden information in an appropriate multimedia carrier, e.g., image, audio, or video. There are several known methods of solving this problem, which operate either in the space domain or in the frequency domain, and are distinguished by the following characteristics: payload, robustness and strength. The payload is the amount of secret data that can be embedded in the carrier without inducing suspicious artefacts, robustness indicates the degree in which the secret data is affected by the normal processing of the carrier e.g., compression, and the strength indicate how easy the presence of hidden data can be detected by steganalysis techniques. This paper presents a new method of hiding secret data into a digital image compressed by a technique based on the Discrete Wavelet Transform (DWT [2] and the Set Partitioning In Hierarchical Trees (SPIHT subband coding algorithm [6]. The proposed method admits huge payloads and has considerable strength.

  16. Lubiprostone Stimulates Duodenal Bicarbonate Secretion in Rats

    OpenAIRE

    Mizumori, Misa; Akiba, Yasutada; Kaunitz, Jonathan D

    2009-01-01

    Background Lubiprostone, a bicyclic fatty acid, is used for the treatment of chronic constipation. No published study has addressed the effect of lubiprostone on intestinal ion secretion in vivo. Aim The aim of this study was to test the hypothesis that lubiprostone augments duodenal HCO3 ? secretion (DBS). Methods Rat proximal duodenal loops were perfused with pH 7.0 Krebs, control vehicle (medium-chain triglycerides), or lubiprostone (0.1?10??M). We measured DBS with flow-through pH and CO2...

  17. REM sleep: tear secretion and dreams.

    Science.gov (United States)

    Murube, Juan

    2008-01-01

    Although a number of hypotheses exist to explain the reasons for the rapid eye movement (REM) phase of sleep, the physical movements themselves have not been explained or interpreted in the literature. The author theorizes that REM during sleep serves not only to increase the lacrimal secretion and to humidify and lubricate the ocular surface, but also to redistribute the secretion on the ocular surface and to inform the conjunctiva-associated lymphoid tissue (CALT) system. He hypothesizes that when eyes move in REM periods to humidify the ocular surface, they indirectly release phenomena of the visual activity, producing dreams.

  18. Factors influencing insulin secretion from encapsulated islets

    NARCIS (Netherlands)

    de Haan, BJ; Faas, MM; de Vos, P

    2003-01-01

    Adequate regulation of glucose levels by a microencapsulated pancreatic islet graft requires a minute-to-minute regulation of blood glucose. To design such a transplant, it is mandatory to have sufficient insight in factors influencing the kinetics of insulin secretion by encapsulated islets. The

  19. Vesicular secretion of auxin: Evidences and implications.

    Science.gov (United States)

    Baluska, Frantisek; Schlicht, Markus; Volkmann, Dieter; Mancuso, Stefano

    2008-04-01

    The plant hormone auxin is secreted in root apices via phospholipase Dzeta2 (PLDzeta2) activity which produces specific population of phosphatidic acid that stimulates secretion of vesicles enriched with auxin. These vesicles were reported to be localized at plant synapses which are active in auxin secretion, especially at the transition zone of the root apex. There are several implications of this vesicular secretion of auxin. In root apices, auxin emerges as plant neurotransmitter-like signal molecule which coordinates activities of adjacent cells via electric and chemical signaling. Putative quantal release of auxin after electrical stimulation, if confirmed, would be part of neuronal communication between plant cells. As auxin transport across plant synapses is tightly linked with integrated sensory perception of environment, especially of omnipresent gravity and light, this process is proposed to mediate the plant perception of environment. These neuronal features allow sessile plants to integrate multitude of sensory signals into the adaptive behavior of whole plants and the animal-like exploratory behavior of growing roots.

  20. Secret Shopping as User Experience Assessment Tool

    Science.gov (United States)

    Boyce, Crystal M.

    2015-01-01

    Secret shopping is a form of unobtrusive evaluation that can be accomplished with minimal effort, but still produce rich results. With as few as 11 shoppers, the author was able to identify trends in user satisfaction with services provided across two entry-level desks at Illinois Wesleyan University's The Ames Library. The focus of this secret…

  1. Removal of Duodenum Elicits GLP-1 Secretion

    DEFF Research Database (Denmark)

    Muscogiuri, Giovanna; Mezza, Teresa; Prioletta, Annamaria

    2013-01-01

    OBJECTIVETo evaluate the effect of removal of the duodenum on the complex interplay between incretins, insulin, and glucagon in nondiabetic subjects.RESEARCH DESIGN AND METHODSFor evaluation of hormonal secretion and insulin sensitivity, 10 overweight patients without type 2 diabetes (age 61 ± 19...

  2. Apical secretion of apolipoproteins from enterocytes

    DEFF Research Database (Denmark)

    Danielsen, E M; Hansen, Gert Helge; Poulsen, Mona Dam

    1993-01-01

    that enterocytes release most of their newly made free apo A-1 and a significant portion of apo B-48 by exocytosis via the brush border membrane into the intestinal lumen. Fat absorption reduced apolipoprotein secretion to the medium and induced the formation of chylomicrons, containing apo A-1 at their surface...

  3. Gastric Acid Secretion, Mucus Concentration and Ulceration ...

    African Journals Online (AJOL)

    This study was carried out to evaluate the effect of consumption (ingestion) of Cannabis sativa on the gastrointestinal tract using mucus concentration, acid secretion and ulceration in animal (rats) model as indices. Three groups of six (6) rats each were used. The control group were fed on rat chow only while another group ...

  4. Experimental Study on Gastric Juice Secretion by ...

    African Journals Online (AJOL)

    管理平台

    2012-05-29

    May 29, 2012 ... reduced (P < 0.05) when acupuncture at zusanli was applied after treatment with cimetidine. Therefore, our study shows that when electroacupuncture at zusanli is applied, the gastric electrical frequency increased and gastric electrical amplitude reduced, while the flux of gastric juice secretion increased.

  5. Secretion Of Methionine By Microorganisms Associated With ...

    African Journals Online (AJOL)

    The organisms were assessed for optimum methionine production at various levels of glucose, ammonium sulphate and varying mixtures of potassium hydrogen phosphate and di-potassium hydrogen phosphate. All the organisms required 10 g glucose for maximum methionine secretion. All the isolates required 20 g of ...

  6. Proprotein Convertases Process Pmel17 during Secretion*

    Science.gov (United States)

    Leonhardt, Ralf M.; Vigneron, Nathalie; Rahner, Christoph; Cresswell, Peter

    2011-01-01

    Pmel17 is a melanocyte/melanoma-specific protein that traffics to melanosomes where it forms a fibrillar matrix on which melanin gets deposited. Before being cleaved into smaller fibrillogenic fragments the protein undergoes processing by proprotein convertases, a class of serine proteases that typically recognize the canonical motif RX(R/K)R↓. The current model of Pmel17 maturation states that this processing step occurs in melanosomes, but in light of recent reports this issue has become controversial. We therefore addressed this question by thoroughly assessing the processing kinetics of either wild-type Pmel17 or a secreted soluble Pmel17 derivative. Our results demonstrate clearly that processing of Pmel17 occurs during secretion and that it does not require entry of the protein into the endocytic system. Strikingly, processing proceeds even in the presence of the secretion inhibitor monensin, suggesting that Pmel17 is an exceptionally good substrate. In line with this, we find that newly synthesized surface Pmel17 is already quantitatively cleaved. Moreover, we demonstrate that Pmel17 function is independent of the sequence identity of its unconventional proprotein convertase-cleavage motif that lacks arginine in P4 position. The data alter the current view of Pmel17 maturation and suggest that the multistep processing of Pmel17 begins with an early cleavage during secretion that primes the protein for later functional processing. PMID:21247888

  7. Proprotein convertases process Pmel17 during secretion.

    Science.gov (United States)

    Leonhardt, Ralf M; Vigneron, Nathalie; Rahner, Christoph; Cresswell, Peter

    2011-03-18

    Pmel17 is a melanocyte/melanoma-specific protein that traffics to melanosomes where it forms a fibrillar matrix on which melanin gets deposited. Before being cleaved into smaller fibrillogenic fragments the protein undergoes processing by proprotein convertases, a class of serine proteases that typically recognize the canonical motif RX(R/K)R↓. The current model of Pmel17 maturation states that this processing step occurs in melanosomes, but in light of recent reports this issue has become controversial. We therefore addressed this question by thoroughly assessing the processing kinetics of either wild-type Pmel17 or a secreted soluble Pmel17 derivative. Our results demonstrate clearly that processing of Pmel17 occurs during secretion and that it does not require entry of the protein into the endocytic system. Strikingly, processing proceeds even in the presence of the secretion inhibitor monensin, suggesting that Pmel17 is an exceptionally good substrate. In line with this, we find that newly synthesized surface Pmel17 is already quantitatively cleaved. Moreover, we demonstrate that Pmel17 function is independent of the sequence identity of its unconventional proprotein convertase-cleavage motif that lacks arginine in P4 position. The data alter the current view of Pmel17 maturation and suggest that the multistep processing of Pmel17 begins with an early cleavage during secretion that primes the protein for later functional processing.

  8. The Best kept Secrets In Government.

    Science.gov (United States)

    1996-09-01

    workers. 113 The Best Kept Secrets In Government • Sweatshops are under attack. More than 50 manufacturing firms across the country have com...Employment and Training Administration Eradicating Sweatshops Initiative Fax on Demand Federal Employees’ Compensation Reengineering Project Team

  9. Secretion of phytohemagglutinin by monkey COS cells.

    Science.gov (United States)

    Voelker, T A; Florkiewicz, R Z; Chrispeels, M J

    1986-12-01

    The entire coding region of a gene, which encodes a polypeptide of phytohemagglutinin (PHA-L), obtained from a library of genomic DNA of the common bean Phaseolus vulgaris cv. Greensleeves, was introduced into the SV40 expression vector pJC119. Monkey COS1 cells were transfected with the recombinant clone and the synthesis, glycosylation, and transport of PHA-L studied and compared with the normal processes in bean cotyledons. In the bean, phytohemagglutinin is synthesized on the rough endoplasmic reticulum and transported via the Golgi complex to protein bodies, vacuole-like organelles. Phytohemagglutinin was synthesized and glycosylated at the ER and processed in the Golgi apparatus of the transfected COS1 cells. After passing the Golgi apparatus, PHA-L was slowly secreted into the culture medium (half-time of 3-6 h), a result indicating that the signals for targeting proteins beyond the Golgi apparatus in plant cells are different from those in animal cells. PHA, which is stored in protein bodies in the plant cells, is secreted by animal cells. Tunicamycin inhibited both glycosylation and secretion of PHA by the COS1 cells, a finding indicating an essential role of the oligosaccharides for transport of PHA in these cells in contrast to the situation found in bean cotyledons. PHA, secreted into the culture medium, was partially sensitive to endo H, a result indicating the presence of one high-mannose and one complex oligosaccharide chain, a situation identical to that in beans.

  10. Surfactant secretion and clearance in the newborn

    Energy Technology Data Exchange (ETDEWEB)

    Stevens, P.A.; Wright, J.R.; Clements, J.A. (Univ. of California, San Francisco (USA))

    1989-10-01

    Pregnant rabbits (30 days) were injected intravenously with (3H)choline 8 h before delivery. The fetuses were delivered, and lung lavage and lamellar body phospholipids (PL) were analyzed. Some newborns also received radioactively labeled surfactant intratracheally on delivery and were permitted to breathe. With time, intratracheal label decreased in lavage and appeared in the lamellar body fraction, and intravenous label accumulated in both pools. Using a tracer analysis for non-steady state, we calculated surfactant secretion and clearance rates for the newborn period. Before birth, both rates rose slightly from 1.8 micrograms PL.g body wt-1.h-1 at 6 h before birth to 7.3 at birth. Immediately after birth, secretion rate rose to 37.7 micrograms PL.g body wt-1.h-1. Between 1.5 and 2 h after birth it fell to a minimum of 1.8 micrograms PL.g body wt-1.h-1 and then rose slowly to 6.0 at 12 h. After birth, clearance rate increased less than secretion rate (maximum 24.7 micrograms PL.g body wt-1.h-1 shortly after birth) then followed the same pattern but did not balance secretion rate in the 1st day.

  11. Type V secretion: From biogenesis to biotechnology

    NARCIS (Netherlands)

    van Ulsen, P.; Rahman, S.U.; Jong, W.S.P.; Daleke, M.H.; Luirink, J.

    2014-01-01

    The two membranes of Gram-negative bacteria contain protein machines that have a general function in their assembly. To interact with the extra-cellular milieu, Gram-negatives target proteins to their cell surface and beyond. Many specialized secretion systems have evolved with dedicated

  12. Lubiprostone stimulates duodenal bicarbonate secretion in rats.

    Science.gov (United States)

    Mizumori, Misa; Akiba, Yasutada; Kaunitz, Jonathan D

    2009-10-01

    Lubiprostone, a bicyclic fatty acid, is used for the treatment of chronic constipation. No published study has addressed the effect of lubiprostone on intestinal ion secretion in vivo. The aim of this study was to test the hypothesis that lubiprostone augments duodenal HCO(3) (-) secretion (DBS). Rat proximal duodenal loops were perfused with pH 7.0 Krebs, control vehicle (medium-chain triglycerides), or lubiprostone (0.1-10 microM). We measured DBS with flow-through pH and CO(2) electrodes, perfusate [Cl(-)] with a Cl(-) electrode, and water flux using a non-absorbable ferrocyanide marker. Some rats were pretreated with a potent, selective CFTR antagonist, CFTR(inh)-172 (1 mg/kg, ip), 1 h before experiments. Perfusion of lubiprostone concentration dependently increased DBS, whereas net Cl(-) output and net water output were only increased at 0.1 microM, compared with vehicle. CFTR(inh)-172 reduced lubiprostone (10 microM)-induced DBS increase, whereas net Cl(-) output was also unchanged. Nevertheless, CFTR(inh)-172 reduced basal net water output, which was reversed by lubiprostone. Furthermore, lubiprostone-induced DBS was inhibited by EP4 receptor antagonist, not by an EP1/2 receptor antagonist or by indomethacin pretreatment. In this first study of the effect of lubiprostone on intestinal ion secretion in vivo, lubiprostone stimulated CFTR-dependent DBS without changing net Cl(-) secretion. This effect supports the hypothesis that Cl(-) secreted by CFTR is recycled across the apical membrane by anion exchangers. Recovery of water output during CFTR inhibition suggests that lubiprostone may improve the intestinal phenotype in CF patients. Furthermore, increased DBS suggests that lubiprostone may protect the duodenum from acid-induced injury via EP4 receptor activation.

  13. Regulated Mucin Secretion from Airway Epithelial Cells

    Directory of Open Access Journals (Sweden)

    Kenneth Bruce Adler

    2013-09-01

    Full Text Available Secretory epithelial cells of the proximal airways synthesize and secrete gel-forming polymeric mucins. The secreted mucins adsorb water to form mucus that is propelled by neighboring ciliated cells, providing a mobile barrier which removes inhaled particles and pathogens from the lungs. Several features of the intracellular trafficking of mucins make the airway secretory cell an interesting comparator for the cell biology of regulated exocytosis. Polymeric mucins are exceedingly large molecules (up to 3x10^6 D per monomer whose folding and initial polymerization in the ER requires the protein disulfide isomerase Agr2. In the Golgi, mucins further polymerize to form chains and possibly branched networks comprising more than 20 monomers. The large size of mucin polymers imposes constraints on their packaging into transport vesicles along the secretory pathway. Sugar side chains account for >70% of the mass of mucins, and their attachment to the protein core by O-glycosylation occurs in the Golgi. Mature polymeric mucins are stored in large secretory granules ~1 um in diameter. These are translocated to the apical membrane to be positioned for exocytosis by cooperative interactions among MARCKS, cysteine string protein (CSP, HSP70 and the cytoskeleton. Mucin granules undergo exocytic fusion with the plasma membrane at a low basal rate and a high stimulated rate. Both rates are mediated by a regulated exocytic mechanism as indicated by phenotypes in both basal and stimulated secretion in mice lacking Munc13-2, a sensor of the second messengers calcium and diacylglycerol (DAG. Basal secretion is induced by low levels of activation of P2Y2 purinergic and A3 adenosine receptors by extracellular ATP released in paracrine fashion and its metabolite adenosine. Stimulated secretion is induced by high levels of the same ligands, and possibly by inflammatory mediators as well. Activated receptors are coupled to phospholipase C by Gq, resulting in the

  14. The unconventional secretion of ARMS2.

    Science.gov (United States)

    Kortvely, Elod; Hauck, Stefanie M; Behler, Jennifer; Ho, Nurulain; Ueffing, Marius

    2016-08-01

    Age-related maculopathy susceptibility 2 (ARMS2) is a small (11 kDa), primate-specific protein found in the extracellular matrix of the choroid layer in the eye. Variants in the corresponding genetic locus are highly associated with age-related macular degeneration, a leading cause of blindness in the elderly. So far, the physiological function of ARMS2 has remained enigmatic. It has been demonstrated that ARMS2 is a genuine secreted protein devoid of an N-terminal leader sequence, yet the mechanism how it exits the cells and enters the choroidal matrix is not understood. Here, we show that ARMS2 efficiently recruits lectin chaperones from the cytosol and colocalizes with calnexin-positive and protein disulfide isomerase-negative vesicle-like structures. Site-directed mutagenesis revealed critical elements for this interaction. Mutant forms proving unable to interact with the calnexin/calreticulin system failed secretion. On the other hand, blocking the endoplasmic reticulum to Golgi transport with brefeldin A had no effect on ARMS2 secretion. As we found ARMS2 colocalizing with GRASP65, a marker for unconventional protein secretion, autophagic factors are likely to be key in its export. Interleukin-1ß (IL-1ß) is the most established example of secretory autophagy. Co-expression experiments, however, suggest that the transport of ARMS2 is different from that of IL-1ß. In conclusion, in this work we show that ARMS2 is externalized via an unconventional pathway bypassing Golgi. Its intracellular separation from the classical secretion pathway suggests that the maturation of the protein requires a specific biochemical niche and/or may be needed to impede the premature formation of unwanted protein-protein interactions. © The Author 2016. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  15. Concentrative biliary secretion of ceftriaxone. Inhibition of lipid secretion and precipitation of calcium ceftriaxone in bile.

    Science.gov (United States)

    Xia, Y; Lambert, K J; Schteingart, C D; GU, J J; Hofmann, A F

    1990-08-01

    The hepatic transport of ceftriaxone, a third-generation cephalosporin, was characterized in the rat and hamster; its effect on bile flow and bile acid-induced biliary lipid secretion was also measured. In anesthetized rats with biliary fistulae, the Tmax was about 5 mumol.min-1.kg-1, and in the hamster the Tmax was about 1 mumol.min-1.kg-1. The compound was not biotransformed. At high secretion rates, the concentration of cephalosporin in bile increased to 27 mmol/L, a concentration far exceeding the solubility product of its calcium salt [2 x 10(-6) (mol/L)2], which precipitated from bile. In the rat, ceftriaxone induced choleresis (22 microL/mumol ceftriaxone, the expected value for a dianionic compound). In the isolated perfused rat liver, ceftriaxone had a fractional hepatic extraction rate averaging 3%; the compound was concentratively secreted into bile, the bile-perfusate ratio ranging from 35-250. Ceftriaxone inhibited phospholipid and cholesterol secretion induced by endogenous or exogenous bile acids; the rate of inhibition was linearly proportional to the canalicular secretion rate of ceftriaxone. Hepatic transport of ceftriaxone had no influence on hepatic secretion of ursodeoxycholyltaurine. In contrast, the net hepatic transport of ursodeoxycholic acid, ursodeoxycholyltaurine, or cholyltaurine inhibited ceftriaxone transport in a dose-dependent manner. It is concluded that ceftriaxone and bile acids share a common mechanism for hepatic transport in the rat and also interact in the processes involved in biliary lipid secretion. Biliary secretion of unbiotransformed ceftriaxone occurs at high concentrations; secondary Ca2+ entry results in the formation of supersaturated canalicular bile and subsequent precipitation as a calcium salt in the biliary tract. These data explain the formation of biliary sludge that occurs in patients undergoing high-dose ceftriaxone therapy.

  16. Long-term outcomes of surgery and radiotherapy for secreting and non-secreting pituitary adenoma

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Mi Young; Kim, Jin Hee; Oh, Young Kee; Kim, El [Dongsan Medical Center, Keimyung University School of Medicine, Daegu (Korea, Republic of)

    2016-06-15

    To investigate treatment outcome and long term complication after surgery and radiotherapy (RT) for pituitary adenoma. From 1990 to 2009, 73 patients with surgery and RT for pituitary adenoma were analyzed in this study. Median age was 51 years (range, 25 to 71 years). Median tumor size was 3 cm (range, 1 to 5 cm) with suprasellar (n = 21), cavernous sinus extension (n = 14) or both (n = 5). Hormone secreting tumor was diagnosed in 29 patients; 16 patients with prolactin, 12 patients with growth hormone, and 1 patient with adrenocorticotrophic hormone. Impairment of visual acuity or visual field was presented in 33 patients at first diagnosis. Most patients (n = 64) received RT as postoperative adjuvant setting. Median RT dose was 45 Gy (range, 45 to 59.4 Gy). Median follow-up duration was 8 years (range, 3 to 22 years). In secreting tumors, hormone normalization rate was 55% (16 of 29 patients). For 25 patients with evaluable visual field and visual acuity test, 21 patients (84%) showed improvement of visual disturbance after treatment. The 10-year tumor control rate for non-secreting and secreting adenoma was 100% and 58%, respectively (p < 0.001). Progression free survival rate at 10 years was 98%. Only 1 patient experienced endocrinological recurrence. Following surgery, 60% (n = 44) suffered from pituitary function deficit. Late complication associated with RT was only 1 patient, who developed cataract. Surgery and RT are very effective and safe in hormonal and tumor growth control for secreting and non-secreting pituitary adenoma.

  17. A thyrotropin‑secreting macroadenoma with positive growth ...

    African Journals Online (AJOL)

    2015-02-01

    TSHoma) are rare adenomas presenting with hyperthyroidism due to impaired negative feedback of thyroid hormone on the pituitary and inappropriate TSH secretion. This article presents a case of TSH‑secreting macroadenoma ...

  18. Trichophyton rubrum secreted and membrane-associated carboxypeptidases

    National Research Council Canada - National Science Library

    Zaugg, Christophe; Jousson, Olivier; Léchenne, Barbara; Staib, Peter; Monod, Michel

    2008-01-01

    .... Therefore, secreted proteolytic activity is considered a virulence trait of these fungi. In a medium containing protein as a sole nitrogen and carbon source Trichophyton rubrum secretes a metallocarboxypeptidase (TruMcpA...

  19. Biliary cholesterol secretion : More than a simple ABC

    NARCIS (Netherlands)

    Dikkers, Arne; Tietge, Uwe J. F.

    2010-01-01

    Biliary cholesterol secretion is a process important for 2 major disease complexes, atherosclerotic cardiovascular disease and cholesterol gallstone disease With respect to cardiovascular disease, biliary cholesterol secretion is regarded as the final step for the elimination of cholesterol

  20. A thyrotropin‑secreting macroadenoma with positive growth ...

    African Journals Online (AJOL)

    TSHoma) are rare adenomas presenting with hyperthyroidism due to impaired negative feedback of thyroid hormone on the pituitary and inappropriate TSH secretion. This article presents a case of TSH‑secreting macroadenoma without any ...

  1. Mining secreted proteins that function in pepper fruit development and ripening using a yeast secretion trap (YST)

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Je Min, E-mail: jemin@knu.ac.kr [Department of Plant Science, College of Agriculture and Life Sciences, Seoul National University, Seoul (Korea, Republic of); Department of Horticultural Science, Kyungpook National University, Daegu (Korea, Republic of); Lee, Sang-Jik [Biotechnology Institute, Nongwoo Bio Co, Ltd, Yeoju (Korea, Republic of); Department of Plant Biology, Cornell University, Ithaca, NY (United States); Rose, Jocelyn K.C. [Department of Plant Biology, Cornell University, Ithaca, NY (United States); Yeam, Inhwa [Department of Horticulture and Breeding, Andong National University, Andong (Korea, Republic of); Kim, Byung-Dong [Department of Plant Science, College of Agriculture and Life Sciences, Seoul National University, Seoul (Korea, Republic of)

    2014-04-18

    Highlights: • Yeast secretion trap (YST) is a valuable tool for mining secretome. • A total of 80 secreted proteins are newly identified via YST in pepper fruits. • The secreted proteins are differentially regulated during pepper development and ripening. • Transient GFP-fusion assay and in planta secretion trap can effectively validate the secretion of proteins. - Abstract: Plant cells secrete diverse sets of constitutively- and conditionally-expressed proteins under various environmental and developmental states. Secreted protein populations, or secretomes have multiple functions, including defense responses, signaling, metabolic processes, and developmental regulation. To identify genes encoding secreted proteins that function in fruit development and ripening, a yeast secretion trap (YST) screen was employed using pepper (Capsicum annuum) fruit cDNAs. The YST screen revealed 80 pepper fruit-related genes (CaPFRs) encoding secreted proteins including cell wall proteins, several of which have not been previously described. Transient GFP-fusion assay and an in planta secretion trap were used to validate the secretion of proteins encoded by selected YST clones. In addition, RNA gel blot analyses provided further insights into their expression and regulation during fruit development and ripening. Integrating our data, we conclude that the YST provides a valuable functional genomics tool for the identification of substantial numbers of novel secreted plant proteins that are associated with biological processes, including fruit development and ripening.

  2. Type VI secretion system in Pseudomonas aeruginosa: secretion and multimerization of VgrG proteins.

    Science.gov (United States)

    Hachani, Abderrahman; Lossi, Nadine S; Hamilton, Alexander; Jones, Cerith; Bleves, Sophie; Albesa-Jové, David; Filloux, Alain

    2011-04-08

    Pseudomonas aeruginosa is a Gram-negative bacterium causing chronic infections in cystic fibrosis patients. Such infections are associated with an active type VI secretion system (T6SS), which consists of about 15 conserved components, including the AAA+ ATPase, ClpV. The T6SS secretes two categories of proteins, VgrG and Hcp. Hcp is structurally similar to a phage tail tube component, whereas VgrG proteins show similarity to the puncturing device at the tip of the phage tube. In P. aeruginosa, three T6SSs are known. The expression of H1-T6SS genes is controlled by the RetS sensor. Here, 10 vgrG genes were identified in the PAO1 genome, among which three are co-regulated with H1-T6SS, namely vgrG1a/b/c. Whereas VgrG1a and VgrG1c were secreted in a ClpV1-dependent manner, secretion of VgrG1b was ClpV1-independent. We show that VgrG1a and VgrG1c form multimers, which confirmed the VgrG model predicting trimers similar to the tail spike. We demonstrate that Hcp1 secretion requires either VgrG1a or VgrG1c, which may act independently to puncture the bacterial envelope and give Hcp1 access to the surface. VgrG1b is not required for Hcp1 secretion. Thus, VgrG1b does not require H1-T6SS for secretion nor does H1-T6SS require VgrG1b for its function. Finally, we show that VgrG proteins are required for secretion of a genuine H1-T6SS substrate, Tse3. Our results demonstrate that VgrG proteins are not only secreted components but are essential for secretion of other T6SS substrates. Overall, we emphasize variability in behavior of three P. aeruginosa VgrGs, suggesting that, although very similar, distinct VgrGs achieve specific functions.

  3. Secrets of over-indebted people

    Directory of Open Access Journals (Sweden)

    Valérie BILLAUDEAU

    2013-12-01

    Full Text Available Today, the economic and financial crisis is brought to light and it is now clear that many people are directly impacted by this phenomenon. However, a lot of situations are obviously hidden and in particular those concerning over-indebted people. These people often find that it is difficult to express the hardship they are going through and keep silent because is more comfortable for them. The media also does not tackle this burning issue because the complexity of some situations complicates the message. Therefore, a giant gap has appeared leaving over-indebted people entrapped in their secret. Starting from this hypothesis, this article will examine the results of a research conducted on the over-indebted people (survey on written press, analysis of TV broadcasts, analysis of records related to person in debts and responses of people in debts and the secrets that this phenomenon involves.

  4. Pancreatic bicarbonate secretion involves two proton pumps

    DEFF Research Database (Denmark)

    Novak, Ivana; Wang, Jing; Henriksen, Katrine L.

    2011-01-01

    H(+)/HCO(3)(-) transporters, which depend on gradients created by the Na(+)/K(+)-ATPase. However, the model cannot fully account for high-bicarbonate concentrations, and other active transporters, i.e. pumps, have not been explored. Here we show that pancreatic ducts express functional gastric...... and non-gastric H(+)-K(+)-ATPases. We measured intracellular pH and secretion in small ducts isolated from rat pancreas and showed their sensitivity to H(+)-K(+) pump inhibitors and ion substitutions. Gastric and non-gastric H(+)-K(+) pumps were demonstrated on RNA and protein levels, and pumps were...... localized to the plasma membranes of pancreatic ducts. Quantitative analysis of H(+)/HCO(3)(-) and fluid transport shows that the H(+)-K(+) pumps can contribute to pancreatic secretion in several species. Our results call for revision of the bicarbonate transport physiology in pancreas, and most likely...

  5. Eccrine sweat gland development and sweat secretion.

    Science.gov (United States)

    Cui, Chang-Yi; Schlessinger, David

    2015-09-01

    Eccrine sweat glands help to maintain homoeostasis, primarily by stabilizing body temperature. Derived from embryonic ectoderm, millions of eccrine glands are distributed across human skin and secrete litres of sweat per day. Their easy accessibility has facilitated the start of analyses of their development and function. Mouse genetic models find sweat gland development regulated sequentially by Wnt, Eda and Shh pathways, although precise subpathways and additional regulators require further elucidation. Mature glands have two secretory cell types, clear and dark cells, whose comparative development and functional interactions remain largely unknown. Clear cells have long been known as the major secretory cells, but recent studies suggest that dark cells are also indispensable for sweat secretion. Dark cell-specific Foxa1 expression was shown to regulate a Ca(2+) -dependent Best2 anion channel that is the candidate driver for the required ion currents. Overall, it was shown that cholinergic impulses trigger sweat secretion in mature glands through second messengers - for example InsP3 and Ca(2+) - and downstream ion channels/transporters in the framework of a Na(+) -K(+) -Cl(-) cotransporter model. Notably, the microenvironment surrounding secretory cells, including acid-base balance, was implicated to be important for proper sweat secretion, which requires further clarification. Furthermore, multiple ion channels have been shown to be expressed in clear and dark cells, but the degree to which various ion channels function redundantly or indispensably also remains to be determined. Published 2015. This article is a U.S. Government work and is in the public domain in the USA.

  6. Eosinophil secretion of granule-derived cytokines

    Directory of Open Access Journals (Sweden)

    Lisa A Spencer

    2014-10-01

    Full Text Available Eosinophils are tissue-dwelling leukocytes, present in the thymus, and gastrointestinal and genitourinary tracts of healthy individuals at baseline, and recruited, often in large numbers, to allergic inflammatory foci and sites of active tissue repair. The biological significance of eosinophils is vast and varied. In health, eosinophils support uterine and mammary gland development, and maintain bone marrow plasma cells and adipose tissue alternatively activated macrophages, while in response to tissue insult eosinophils function as inflammatory effector cells, and, in the wake of an inflammatory response, promote tissue regeneration and wound healing. One common mechanism driving many of the diverse eosinophil functions is the regulated and differential secretion of a vast array of eosinophil-derived cytokines. Eosinophils are distinguished from most other leukocytes in that many, if not all, of the over three dozen eosinophil-derived cytokines are pre-synthesized and stored within intracellular granules, poised for very rapid, stimulus-induced secretion. Eosinophils engaged in cytokine secretion in situ utilize distinct pathways of cytokine release that include: classical exocytosis, whereby granules themselves fuse with the plasma membrane and release their entire contents extracellularly; piecemeal degranulation, whereby granule-derived cytokines are selectively mobilized into vesicles that emerge from granules, traverse the cytoplasm and fuse with the plasma membrane to release discrete packets of cytokines; and eosinophil cytolysis, whereby intact granules are extruded from eosinophils, and deposited within tissues. In this latter scenario, extracellular granules can themselves function as stimulus-responsive secretory-competent organelles within the tissue. Here we review the distinctive processes of differential secretion of eosinophil granule-derived cytokines.

  7. Unconventional protein secretion (UPS) pathways in plants.

    Science.gov (United States)

    Ding, Yu; Robinson, David G; Jiang, Liwen

    2014-08-01

    As in yeast and mammalian cells, novel unconventional protein secretion (UPS) or unconventional membrane trafficking pathways are now known to operate in plants. UPS in plants is generally associated with stress conditions such as pathogen attack, but little is known about its underlying mechanism and function. Here, we present an update on the current knowledge of UPS in the plants in terms of its transport pathways, possible functions and its relationship to autophagy. Copyright © 2014 Elsevier Ltd. All rights reserved.

  8. George's secret key to the universe

    CERN Document Server

    Hawking, Lucy; Galfard, Christophe; Parsons, Gary

    2007-01-01

    In their bestselling book for young readers, noted physicist Stephen Hawking and his daughter, Lucy, provide a grand and funny adventure that explains fascinating information about our universe, including Dr. Hawking's latest ideas about black holes. It's the story of George, who's taken through the vastness of space by a scientist, his daughter, and their super-computer named Cosmos. George's Secret Key to the Universe was a New York Times bestseller and a selection of Al's Book Club on the Today show.

  9. ACTH-Secreting Pheochromocytoma. Case report

    Directory of Open Access Journals (Sweden)

    N S Kuznetsov

    2012-12-01

    Full Text Available Ectopic hormone-secreting pheochromocytomas are rare. Only case reports exist in the literature. Despite the large number of guides on diagnosis and treatment of pheochromocytoma, and Cushing syndrome, the extreme rarity of ectopic ACTH-syndrome caused by pheochromocytoma, and complexity of clinical cause numerous diagnostic errors leading to treatment failure. Therefore, we belive it appropriate to share our experience of this group of patients.

  10. Exosomes: secreted vesicles and intercellular communications

    OpenAIRE

    Théry, Clotilde

    2011-01-01

    Exosomes are small membrane vesicles of endocytic origin secreted by most cell types, and are thought to play important roles in intercellular communications. Although exosomes were originally described in 1983, interest in these vesicles has really increased dramatically in the last 3 years, after the finding that they contain mRNA and microRNA. This discovery sparked renewed interest for the general field of membrane vesicles involved in intercellular communications, and research on these s...

  11. A Population Model of Vasopressin Secretion

    OpenAIRE

    Durie, Ruth Frances

    2008-01-01

    Computer modelling is a powerful tool for clarifying and testing theory. In neuroscience, this often means replicating firing patterns. Models need evaluation functions to quantify the significance of features in the firing patterns, but usually the effect of firing is insufficiently understood. The magnocellular vasopressin neurons of the hypothalamus do have an output that is both well understood and quantifiable: they secrete a hormone into the bloodstream in proportion to blood osmol...

  12. Microbial cell surfaces and secretion systems

    OpenAIRE

    Tommassen, J.P.M.; Wosten, H.A.B.

    2015-01-01

    Microbial cell surfaces, surface-exposed organelles, and secreted proteins are important for the interaction with the environment, including adhesion to hosts, protection against host defense mechanisms, nutrient acquisition, and intermicrobial competition. Here, we describe the structures of the cell envelopes of bacteria , fungi, and oomycetes , and the mechanisms they have evolved for the transport of proteins across these envelopes to the cell surface and into the extracellular milieu.

  13. Hoopoes color their eggs with antimicrobial uropygial secretions

    Science.gov (United States)

    Soler, Juan J.; Martín-Vivaldi, M.; Peralta-Sánchez, J. M.; Arco, L.; Juárez-García-Pelayo, N.

    2014-09-01

    Uropygial gland secretions are used as cosmetics by some species of birds to color and enhance properties of feathers and teguments, which may signal individual quality. Uropygial secretions also reach eggshells during incubation and, therefore, may influence the coloration of birds' eggs, a trait that has attracted the attention of evolutionary biologists for more than one century. The color of hoopoe eggs typically changes along incubation, from bluish-gray to greenish-brown. Here, we test experimentally the hypothesis that dark uropygial secretion of females is responsible for such drastic color change. Moreover, since uropygial secretion of hoopoes has antimicrobial properties, we also explore the association between color and antimicrobial activity of the uropygial secretion of females. We found that eggs stayed bluish-gray in nests where female access to the uropygial secretion was experimentally blocked. Furthermore, experimental eggs that were maintained in incubators and manually smeared with uropygial secretion experienced similar color changes that naturally incubated eggs did, while control eggs that were not in contact with the secretions did not experience such color changes. All these results strongly support the hypothesis that female hoopoes use their uropygial gland secretion to color the eggs. Moreover, saturation of the uropygial secretion was associated with antimicrobial activity against Bacillus licheniformis. Given the known antimicrobial potential of uropygial secretions of birds, this finding opens the possibility that in scenarios of sexual selection, hoopoes in particular and birds in general signal antimicrobial properties of their uropygial secretion by mean of changes in egg coloration along incubation.

  14. 10 CFR 1016.34 - Accountability for Secret Restricted Data.

    Science.gov (United States)

    2010-01-01

    ... 10 Energy 4 2010-01-01 2010-01-01 false Accountability for Secret Restricted Data. 1016.34 Section... Information § 1016.34 Accountability for Secret Restricted Data. Each permittee possessing documents containing Secret Restricted Data shall establish a document accountability procedure and shall maintain...

  15. Preconditioning stimuli that augment chromaffin cell secretion.

    Science.gov (United States)

    Tapia, Laura; García-Eguiagaray, Josefina; García, Antonio G; Gandía, Luis

    2009-04-01

    We have investigated here whether a preconditioned stimulation of nicotinic and muscarinic receptors augmented the catecholamine release responses elicited by supramaximal 3-s pulses of 100 muM acetylcholine (100ACh) or 100 mM K(+) (100K(+)) applied to fast-perifused bovine adrenal chromaffin cells. Threshold concentrations of nicotine (1-3 muM) that caused only a tiny secretion did, however, augment the responses elicited by 100ACh or 100K(+) by 2- to 3.5-fold. This effect was suppressed by mecamylamine and by Ca(2+) deprivation, was developed with a half-time (t(1/2)) of 1 min, and was reversible. The nicotine effect was mimicked by threshold concentrations of ACh, choline, epibatidine, and oxotremorine-M but not by methacholine. Threshold concentrations of K(+) caused lesser potentiation of secretion compared with that of threshold nicotine. The data are compatible with an hypothesis implying 1) that continuous low-frequency sympathetic discharge places chromaffin cells at the adrenal gland in a permanent "hypersensitive" state; and 2) this allows an explosive secretion of catecholamines by high-frequency sympathetic discharge during stress.

  16. Secretion of dilevalol in breast milk.

    Science.gov (United States)

    Radwanski, E; Nagabhushan, N; Affrime, M B; Perentesis, G; Symchowicz, S; Patrick, J E

    1988-05-01

    The pharmacokinetics of unchanged and total (unchanged plus Glusulase [Biotechnology Systems, Boston, MA]) released dilevalol and secretion into human breast milk was studied in six healthy breast-feeding female volunteers administered a single 400-mg dilevalol hydrochloride capsule. In plasma, the mean Cmax for unchanged dilevalol, 485 ng/mL was reached at 0.8 hour (tmax) and the AUC(48 hours) was 1435 hr X ng/mL. Pharmacokinetic analysis of unchanged dilevalol in plasma showed that dilevalol was distributed and eliminated with half-lives of 0.9 and 8.2 hours, respectively. Breast milk concentrations of unchanged dilevalol as a function of time, paralleled those of plasma but were consistently lower. The milk Cmax, 149 ng/mL, occurred during the 0 to 2 hour collection interval; the AUC(42 hours) for unchanged dilevalol in milk was 663 hr X ng/mL. The mean milk to plasma concentration ratio was 0.46. The unchanged dilevalol plasma concentrations were 12 to 18% those of total drug suggesting that the drug is extensively conjugated. By contrast, the concentrations of unchanged dilevalol in breast milk, based on Cmax and AUC data were 63 to 94% those of total drug, indicating that very little conjugated drug is secreted into breast milk. Through 48 hours, a mean of only 27 micrograms dilevalol or 0.007% of the administered dose was secreted into breast milk, which is much less than that reported for other beta blockers.

  17. Unconventional Protein Secretion in Animal Cells.

    Science.gov (United States)

    Ng, Fanny; Tang, Bor Luen

    2016-01-01

    All eukaryotic cells secrete a range of proteins in a constitutive or regulated manner through the conventional or canonical exocytic/secretory pathway characterized by vesicular traffic from the endoplasmic reticulum, through the Golgi apparatus, and towards the plasma membrane. However, a number of proteins are secreted in an unconventional manner, which are insensitive to inhibitors of conventional exocytosis and use a route that bypasses the Golgi apparatus. These include cytosolic proteins such as fibroblast growth factor 2 (FGF2) and interleukin-1β (IL-1β), and membrane proteins that are known to also traverse to the plasma membrane by a conventional process of exocytosis, such as α integrin and the cystic fibrosis transmembrane conductor (CFTR). Mechanisms underlying unconventional protein secretion (UPS) are actively being analyzed and deciphered, and these range from an unusual form of plasma membrane translocation to vesicular processes involving the generation of exosomes and other extracellular microvesicles. In this chapter, we provide an overview on what is currently known about UPS in animal cells.

  18. Dragon's blood secretion and its ecological significance.

    Science.gov (United States)

    Jura-Morawiec, Joanna; Tulik, Mirela

    Dragon's blood is the name given to a red exudate produced by some plant species belonging to the genera Daemonorops, Dracaena, Croton and Pterocarpus. These are endemic to various parts of the globe. It is classified as a resin or latex depending on its mode of secretion and its chemical composition, which is species specific. This red substance functions in defence and is produced (a) constitutively and stored in preformed anatomical structures, or (b) by induction in response to traumatic events, such as mechanical injury, pathogen attack or invasion by insects. Apart from its defensive role in plants, dragon's blood is also a valuable natural resource renowned since antiquity for its diverse medicinal properties and uses in art. Despite the great importance of dragon's blood, our knowledge of the biological basis for its secretion is still incomplete. This review summarizes recent advances in the study of the anatomical basis for its secretion, and discusses its classification and ecological function. Bringing some clarity to these issues may also help in the commercial sourcing of dragon's blood.

  19. An Ectopic ACTH Secreting Metastatic Parotid Tumour

    Directory of Open Access Journals (Sweden)

    Thomas Dacruz

    2016-01-01

    Full Text Available A 60-year old woman presented with features of Cushing’s syndrome (CS secondary to an ectopic adrenocorticotropic hormone (ACTH secreting metastatic parotid tumour 3 years after excision of the original tumour. She subsequently developed fatal intestinal perforation and unfortunately died despite best possible medical measures. Ectopic ACTH secretion accounts for 5–10% of all patients presenting with ACTH dependent hypercortisolism; small cell carcinoma of lung (SCLC and neuroendocrine tumours (NET account for the majority of such cases. Although there are 4 previous case reports of ectopic ACTH secreting salivary tumours in literature, to our knowledge this is the first published case report in which the CS developed after 3 years of what was deemed as a successful surgical excision of primary salivary tumour. Our patient initially had nonspecific symptoms which may have contributed to a delay in diagnosis. Perforation of sigmoid colon is a recognised though underdiagnosed complication associated with steroid therapy and hypercortisolism. This case demonstrates the challenges faced in diagnosis as well as management of patients with CS apart from the practical difficulties faced while trying to identify source of ectopic ACTH.

  20. Tubular control of renin synthesis and secretion

    Science.gov (United States)

    Briggs, Josephine P.

    2012-01-01

    The intratubular composition of fluid at the tubulovascular contact site of the juxtaglomerular apparatus serves as regulatory input for secretion and synthesis of renin. Experimental evidence, mostly from in vitro perfused preparations, indicates an inverse relation between luminal NaCl concentration and renin secretion. The cellular transduction mechanism is initiated by concentration-dependent NaCl uptake through the Na–K–2Cl cotransporter (NKCC2) with activation of NKCC2 causing inhibition and deactivation of NKCC2 causing stimulation of renin release. Changes in NKCC2 activity are coupled to alterations in the generation of paracrine factors that interact with granular cells. Among these factors, generation of PGE2 in a COX-2-dependent fashion appears to play a dominant role in the stimulatory arm of tubular control of renin release. [NaCl] is a determinant of local PG release over an appropriate concentration range, and blockade of COX-2 activity interferes with the NaCl dependency of renin secretion. The complex array of local paracrine controls also includes nNOS-mediated synthesis of nitric oxide, with NO playing the role of a modifier of the intracellular signaling pathway. A role of adenosine may be particularly important when [NaCl] is increased, and at least some of the available evidence is consistent with an important suppressive effect of adenosine at higher salt concentrations. PMID:22665048

  1. sizzled function and secreted factor network dynamics

    Directory of Open Access Journals (Sweden)

    Jianli Shi

    2012-02-01

    Studies on the role of the E-box binding transcription factor Snail2 (Slug in the induction of neural crest by mesoderm (Shi et al., 2011 revealed an unexpected increase in the level of sizzled RNA in the dorsolateral mesodermal zone (DMLZ of morphant Xenopus embryos. sizzled encodes a secreted protein with both Wnt and BMP inhibitor activities. Morpholino-mediated down-regulation of sizzled expression in one cell of two cell embryos or the C2/C3 blastomeres of 32-cell embryos, which give rise to the DLMZ, revealed decreased expression of the mesodermal marker brachyury and subsequent defects in neural crest induction, pronephros formation, and muscle patterning. Loss of sizzled expression led to decreases in RNAs encoding the secreted Wnt inhibitor SFRP2 and the secreted BMP inhibitor Noggin; the sizzled morphant phenotype could be rescued by co-injection of RNAs encoding Noggin and either SFRP2 or Dickkopf (a mechanistically distinct Wnt inhibitor. Together, these observations reveal that sizzled, in addition to its established role in dorsal-ventral patterning, is also part of a dynamic BMP and Wnt signaling network involved in both mesodermal patterning and neural crest induction.

  2. Adopting Frank Warren's PostSecret Art Project to Illustrate the Role of Secrets in Interpersonal Communication

    Science.gov (United States)

    Paxman, Christina G.

    2013-01-01

    The exploration of secrets summons the adage that ''what someone doesn't know won't hurt them.'' While this phrase implies that keeping secrets can be advantageous, it also foreshadows another consideration: secrets have the propensity to hurt others (Caughlin, Scott, Miller, & Hefner, 2009). Despite this, the act…

  3. Selective identification of secreted and transmembrane breast cancer markers using Escherichia coli ampicillin secretion trap.

    Science.gov (United States)

    Ferguson, Deborah A; Muenster, Matthew R; Zang, Qun; Spencer, Jeffrey A; Schageman, Jeoffrey J; Lian, Yun; Garner, Harold R; Gaynor, Richard B; Huff, J Warren; Pertsemlidis, Alexander; Ashfaq, Raheela; Schorge, John; Becerra, Carlos; Williams, Noelle S; Graff, Jonathan M

    2005-09-15

    Secreted and cell surface proteins play important roles in cancer and are potential drug targets and tumor markers. Here, we describe a large-scale analysis of the genes encoding secreted and cell surface proteins in breast cancer. To identify these genes, we developed a novel signal sequence trap method called Escherichia coli ampicillin secretion trap (CAST). For CAST, we constructed a plasmid in which the signal sequence of beta-lactamase was deleted such that it does not confer ampicillin resistance. Eukaryotic cDNA libraries cloned into pCAST produced tens of thousands of ampicillin-resistant clones, 80% of which contained cDNA fragments encoding secreted and membrane spanning proteins. We identified 2,708 unique sequences from cDNA libraries made from surgical breast cancer specimens. We analyzed the expression of 1,287 of the 2,708 genes and found that 166 were overexpressed in breast cancers relative to normal breast tissues. Eighty-five percent of these genes had not been previously identified as markers of breast cancer. Twenty-three of the 166 genes (14%) were relatively tissue restricted, suggesting use as cancer-specific targets. We also identified several new markers of ovarian cancer. Our results indicate that CAST is a robust, rapid, and low cost method to identify cell surface and secreted proteins and is applicable to a variety of relevant biological questions.

  4. EFFECTS OF SECRETABLE PLACENTAL FACTORS UPON SECRETION OF CYTOKINES BY THP-1 MONOCYTE-LIKE CELLS

    Directory of Open Access Journals (Sweden)

    Ya. S. Onokhina

    2013-01-01

    Full Text Available Abstract. Мonocytes in feto-placental circulation are exposed to factors secreted by placental tissue. These factors influence monocyte functions in pregnancy. In present study, an in vitro model (monocyte-like THP-1 cells was used for assessing effects of soluble placental factors obtained from women with physiological pregnancies, or preeclampsia cases. The following effects of placental factors were revealed: increased secretion of VEGF by THP-1 cells along with decreased secretion of IL-6, IL-8 and MCP-1 under the influence of placental factors from the I. trimester of pregnancy in comparison with III. trimester. Secretion of IL-6 and MCP-1 by THP-1 cells was increased, and secretion of soluble TNFRII was decreased upon co-cultivation with soluble placental factors from the women with preeclampsia, as compared with placental products from physiological pregnancies.The work is supported by grants ГК № 02.740.11.0711 from Ministry of Education and Science, and НШ-3594.2010.7 grant from the President of Russian Federation.

  5. The role of jasmonates in floral nectar secretion.

    Directory of Open Access Journals (Sweden)

    Venkatesan Radhika

    Full Text Available Plants produce nectar in their flowers as a reward for their pollinators and most of our crops depend on insect pollination, but little is known on the physiological control of nectar secretion. Jasmonates are well-known for their effects on senescence, the development and opening of flowers and on plant defences such as extrafloral nectar. Their role in floral nectar secretion has, however, not been explored so far. We investigated whether jasmonates have an influence on floral nectar secretion in oil-seed rape, Brassica napus. The floral tissues of this plant produced jasmonic acid (JA endogenously, and JA concentrations peaked shortly before nectar secretion was highest. Exogenous application of JA to flowers induced nectar secretion, which was suppressed by treatment with phenidone, an inhibitor of JA synthesis. This effect could be reversed by additional application of JA. Jasmonoyl-isoleucine and its structural mimic coronalon also increased nectar secretion. Herbivory or addition of JA to the leaves did not have an effect on floral nectar secretion, demonstrating a functional separation of systemic defence signalling from reproductive nectar secretion. Jasmonates, which have been intensively studied in the context of herbivore defences and flower development, have a profound effect on floral nectar secretion and, thus, pollination efficiency in B. napus. Our results link floral nectar secretion to jasmonate signalling and thereby integrate the floral nectar secretion into the complex network of oxylipid-mediated developmental processes of plants.

  6. Tracking Proteins Secreted by Bacteria: What's in the Toolbox?

    Directory of Open Access Journals (Sweden)

    Benoit Maffei

    2017-05-01

    Full Text Available Bacteria have acquired multiple systems to expose proteins on their surface, release them in the extracellular environment or even inject them into a neighboring cell. Protein secretion has a high adaptive value and secreted proteins are implicated in many functions, which are often essential for bacterial fitness. Several secreted proteins or secretion machineries have been extensively studied as potential drug targets. It is therefore important to identify the secretion substrates, to understand how they are specifically recognized by the secretion machineries, and how transport through these machineries occurs. The purpose of this review is to provide an overview of the biochemical, genetic and imaging tools that have been developed to evaluate protein secretion in a qualitative or quantitative manner. After a brief overview of the different tools available, we will illustrate their advantages and limitations through a discussion of some of the current open questions related to protein secretion. We will start with the question of the identification of secreted proteins, which for many bacteria remains a critical initial step toward a better understanding of their interactions with the environment. We will then illustrate our toolbox by reporting how these tools have been applied to better understand how substrates are recognized by their cognate machinery, and how secretion proceeds. Finally, we will highlight recent approaches that aim at investigating secretion in real time, and in complex environments such as a tissue or an organism.

  7. Reelin secreted by GABAergic neurons regulates glutamate receptor homeostasis.

    Directory of Open Access Journals (Sweden)

    Cecilia Gonzalez Campo

    Full Text Available BACKGROUND: Reelin is a large secreted protein of the extracellular matrix that has been proposed to participate to the etiology of schizophrenia. During development, reelin is crucial for the correct cytoarchitecture of laminated brain structures and is produced by a subset of neurons named Cajal-Retzius. After birth, most of these cells degenerate and reelin expression persists in postnatal and adult brain. The phenotype of neurons that bind secreted reelin and whether the continuous secretion of reelin is required for physiological functions at postnatal stages remain unknown. METHODOLOGY/PRINCIPAL FINDINGS: Combining immunocytochemical and pharmacological approaches, we first report that two distinct patterns of reelin expression are present in cultured hippocampal neurons. We show that in hippocampal cultures, reelin is secreted by GABAergic neurons displaying an intense reelin immunoreactivity (IR. We demonstrate that secreted reelin binds to receptors of the lipoprotein family on neurons with a punctate reelin IR. Secondly, using calcium imaging techniques, we examined the physiological consequences of reelin secretion blockade. Blocking protein secretion rapidly and reversibly changes the subunit composition of N-methyl-D-aspartate glutamate receptors (NMDARs to a predominance of NR2B-containing NMDARs. Addition of recombinant or endogenously secreted reelin rescues the effects of protein secretion blockade and reverts the fraction of NR2B-containing NMDARs to control levels. Therefore, the continuous secretion of reelin is necessary to control the subunit composition of NMDARs in hippocampal neurons. CONCLUSIONS/SIGNIFICANCE: Our data show that the heterogeneity of reelin immunoreactivity correlates with distinct functional populations: neurons synthesizing and secreting reelin and/or neurons binding reelin. Furthermore, we show that continuous reelin secretion is a strict requirement to maintain the composition of NMDARs. We propose

  8. Proteomics of protein secretion by Aggregatibacter actinomycetemcomitans.

    Directory of Open Access Journals (Sweden)

    Vincent Zijnge

    Full Text Available The extracellular proteome (secretome of periodontitis-associated bacteria may constitute a major link between periodontitis and systemic diseases. To obtain an overview of the virulence potential of Aggregatibacter actinomycetemcomitans, an oral and systemic human pathogen implicated in aggressive periodontitis, we used a combined LC-MS/MS and bioinformatics approach to characterize the secretome and protein secretion pathways of the rough-colony serotype a strain D7S. LC-MS/MS revealed 179 proteins secreted during biofilm growth. Further to confirming the release of established virulence factors (e.g. cytolethal distending toxin [CDT], and leukotoxin [LtxA], we identified additional putative virulence determinants in the secretome. These included DegQ, fHbp, LppC, Macrophage infectivity protein (MIP, NlpB, Pcp, PotD, TolB, and TolC. This finding indicates that the number of extracellular virulence-related proteins is much larger than previously demonstrated, which was also supported by in silico analysis of the strain D7S genome. Moreover, our LC-MS/MS and in silico data revealed that at least Type I, II, and V secretion are actively used to excrete proteins directly into the extracellular space, or via two-step pathways involving the Sec/Tat systems for transport across the inner membrane, and outer membrane factors, secretins and auto-transporters, respectively for delivery across the outer membrane. Taken together, our results provide a molecular basis for further elucidating the role of A. actinomycetemcomitans in periodontal and systemic diseases.

  9. Secret Objective Standoff: International Safeguards Educational Exercise

    Energy Technology Data Exchange (ETDEWEB)

    Okowita, Samantha L [ORNL

    2014-01-01

    The International Safeguards Regime, being so multi-faceted, can be overwhelming to those first introduced to its many components. The organizers and lecturers of workshops and courses on nonproliferation often provide a series of independent lectures and must somehow demonstrate the cohesive and effective nature of the system. An exercise titled The Secret Objective Standoff was developed to complement lectures with hands-on learning to assist participants in bringing all the many components (IAEA agreements, export controls, treaty obligations, international diplomacy, etc.) of the International Safeguards Regime together. This exercise divides participants into teams that are assigned the role of either a country or the IAEA and asks that they fully immerse themselves in their roles. The teams are then randomly assigned three unique and secret objectives that are intended to represent realistic and current geopolitical scenarios. Through construction, trading, or hoarding of four resources (experts, technology, money, and uranium), the teams have a finite number of turns to accomplish their objectives. Each turn has three phases random dispersal of resources, a timed discussion where teams can coordinate and strategize with others, and an action phase. During the action phase, teams inform the moderator individually and secretly what they will be doing that turn. The exercise has been tested twice with Oak Ridge National Laboratory personnel, and has been conducted with outside participants twice, in each case the experience was well received by both participants and instructors. This exercise provides instructors the ability to modify the exercise before or during game play to best fit their educational goals. By offering a range of experiences, from an in-depth look at specific components to a generalized overview, this exercise is an effective tool in helping participants achieve a full understanding the International Safeguards Regime.

  10. [Ear-acupuncture and endocrine secretion].

    Science.gov (United States)

    Spagnoletti, T; Liotti, M; Tutino, L; Marcucci, F; Cafaro, M R; Officioso, A; De Luca, R

    1982-03-15

    We have investigated the possible influence of auricolar puncture over hormonal secretion. We have studied in 5 subjects the course of hormonal incretion determinable by only auricolar puncture and the second time making the same methodology united with the test of hypophyseal stimulation by "LH-RH-insulina-TRH". The results show that it is not get any changes of the plasmatic rates of hypophiseal hormones, while we have observed a decrease of the plasmatic hydrocortisone rate when it was practised the only auricolar puncture.

  11. Proton pump inhibitors inhibit pancreatic secretion

    DEFF Research Database (Denmark)

    Wang, Jing; Barbuskaite, Dagne; Tozzi, Marco

    2015-01-01

    +/K+-ATPases are expressed and functional in human pancreatic ducts and whether proton pump inhibitors (PPIs) have effect on those. Here we show that the gastric HKα1 and HKβ subunits (ATP4A; ATP4B) and non-gastric HKα2 subunits (ATP12A) of H+/K+-ATPases are expressed in human pancreatic cells. Pumps have similar...... localizations in duct cell monolayers (Capan-1) and human pancreas, and notably the gastric pumps are localized on the luminal membranes. In Capan-1 cells, PPIs inhibited recovery of intracellular pH from acidosis. Furthermore, in rats treated with PPIs, pancreatic secretion was inhibited but concentrations...

  12. Secrets Pages in DR’s Playbook

    DEFF Research Database (Denmark)

    Nielsen, Jakob Isak

    According to former Head of Drama at DR, Ingolf Gabold, one of the “great” and “strange” secrets subtending the success of DR’s TV series is the extremely deliberate work with audiovisual style (Gabold in Nielsen 2012). Here - as elsewhere - Gabold highlights the so-called “paramount...... aspect of DR’s productions by interviewing both management and creative personnel) this paper will discuss the causal mechanisms that subtend the development of audiovisual style in DR series from Gøngehøvdingen to 1864 and will conclude with reflections on the role of DR-series’ style in the age...

  13. Proteases induce secretion of collagenase and plasminogen activator by fibroblasts

    Energy Technology Data Exchange (ETDEWEB)

    Werb, Z.; Aggeler, J.

    1978-04-01

    We have observed that treatment of rabbit synovial fibroblasts with proteolytic enzymes can induce secretion of collagenase (EC 3.4.24.7) and plasminogen activator (EC 3.4.21.-). Cells treated for 2 to 24 hr with plasmin, trypsin, chymotrypsin, pancreatic elastase, papain, bromelain, thermolysin, or ..cap alpha..-protease but not with thrombin or neuraminidase secreted detectable amounts of collagenase within 16 to 48 hr. Treatment of fibroblasts with trypsin also induced secretion of plasminogen activator. Proteases initiated secretion of collagenase (up to 20 units per 10/sup 6/ cells per 24 hr) only when treatment produced decreased cell adhesion. Collagenase production did not depend on continued presence of proteolytic activity or on subsequent cell adhesion, spreading, or proliferation. Routine subculturing with crude trypsin also induced collagenase secretion by cells. Secretion of collagenase was prevented and normal spreading was obtained if the trypsinized cells were placed into medium containing fetal calf serum. Soybean trypsin inhibitor, ..cap alpha../sub 1/-antitrypsin, bovine serum albumin, collagen, and fibronectin did not inhibit collagenase production. Although proteases that induced collagenase secretion also removed surface glycoprotein, the kinetics of induction of cell protease secretion were different from those for removal of fibronectin. Physiological inducers of secretion of collagenase and plasminogen activator by cells have not been identified. These results suggest that extracellular proteases in conjunction with plasma proteins may govern protease secretion by cells.

  14. Dopamine-Secreting Paraganglioma in the Retroperitoneum.

    Science.gov (United States)

    Matsuda, Yusuke; Kimura, Noriko; Yoshimoto, Takanobu; Sekiguchi, Yoshihiro; Tomoishi, Junzo; Kasahara, Ichiro; Hara, Yoshihito; Ogawa, Yoshihiro

    2017-03-01

    Pheochromocytomas and paragangliomas, which exclusively produce dopamine, are very rare. Herein, we report for the first time a Japanese case of an exclusively dopamine-producing paraganglioma accompanied by detailed immunohistochemical analyses. A 70-year-old Japanese woman was referred to our hospital for functional examination of her left retroperitoneal mass. Her adrenal functions were normal, except for excessive dopamine secretion. After the tumorectomy, her dopamine level normalized. The histopathological diagnosis of the tumor was paraganglioma; this was confirmed by positive immunostaining of chromogranin A (CgA), tyrosine hydroxylase (TH), dopamine β-hydroxylase (DBH), and succinate dehydrogenase gene subunit B (SDHB). However, the immunostaining of CgA in the tumor cells showed peculiar dot-like staining located corresponding to Golgi complex in the perinuclear area, rather than the diffuse cytoplasmic staining usually observed in epinephrine- or norepinephrine-producing functional pheochromocytomas and paragangliomas. The immunohistochemical results suggested that the tumor cells had sparse neuroendocrine granules in the cytoplasm, resulting in inhibition of catecholamine synthesis from dopamine to norepinephrine in neurosecretory granules. This may be the mechanism responsible for exclusive dopamine secretion in the present case.

  15. Secretion of clostridium cellulase by E. coli

    Science.gov (United States)

    Yu, Ida Kuo

    1998-01-01

    A gene, encoding an endocellulase from a newly isolated mesophilic Clostridium strain IY-2 which can digest bamboo fibers, cellulose, rice straw, and sawdust, was isolated by shotgun cloning in an E. coli expression plasmid pLC2833. E. coli positive clones were selected based on their ability to hydrolyze milled bamboo fibers and cellulose present in agar plates. One clone contained a 2.8 kb DNA fragment that was responsible for cellulase activity. Western blot analyses indicated that the positive clone produced a secreted cellulase with a mass of about 58,000 daltons that was identical in size to the subunit of one of the three major Clostridium cellulases. The products of cellulose digestion by this cloned cellulase were cellotetraose and soluble higher polymers. The cloned DNA contained signal sequences capable of directing the secretion of heterologous proteins from an E. coli host. The invention describes a bioprocess for the treatment of cellulosic plant materials to produce cellular growth substrates and fermentation end products suitable for production of liquid fuels, solvents, and acids.

  16. Management of oral secretions in neurological disease.

    Science.gov (United States)

    McGeachan, Alexander J; Mcdermott, Christopher J

    2017-04-01

    Sialorrhoea is a common and problematic symptom that arises from a range of neurological conditions associated with bulbar or facial muscle dysfunction. Drooling can significantly affect quality of life due to both physical complications such as oral chapping, and psychological complications such as embarrassment and social isolation. Thicker, tenacious oral and pharyngeal secretions may result from the drying management approach to sialorrhoea. The management of sialorrhoea in neurological diseases depends on the underlying pathology and severity of symptoms. Interventions include anticholinergic drugs, salivary gland-targeted radiotherapy, salivary gland botulinum toxin and surgical approaches. The management of thick secretions involves mainly conservative measures such as pineapple juice as a lytic agent, cough assist, saline nebulisers and suctioning or mucolytic drugs like carbocisteine. Despite a current lack of evidence and variable practice, management of sialorrhoea should form a part of the multidisciplinary approach needed for long-term neurological conditions. © Article author(s) (or their employer(s) unless otherwise stated in the text of the article) 2017. All rights reserved. No commercial use is permitted unless otherwise expressly granted.

  17. Gastric Lipase Secretion in Children with Gastritis

    Directory of Open Access Journals (Sweden)

    Krystyna Sztefko

    2013-07-01

    Full Text Available Gastric lipase is one of the prepancreatic lipases found in some mammalian species and in humans. Our knowledge of the hormonal regulation of gastric lipase secretion in children and adolescents is still very limited. The aim of this study was to compare the activity of human gastric lipase (HGL in gastric juice in healthy adolescents and in patients with gastritis. The adolescents were allocated to three groups: the first including patients with Helicobacter pylori gastritis (HPG; n = 10, the second including patients with superficial gastritis caused by pathogens other than H. pylori (non-HPG; n = 14 and the control group including healthy adolescents (n = 14. Activity of HGL was measured in gastric juice collected during endoscopy. Plasma concentrations of cholecystokinin (CCK, glucagon-like peptide-1 (GLP-1 and glucose-dependent insulinotropic peptide (GIP were measured in all adolescents. Activity of HGL in the non-HPG group was significantly lower than in the HPG group (p < 0.005 and the control group (p < 0.005. Mean plasma GIP levels in the control group were lower than in the non-HPG group (p < 0.003 and the HPG group (p < 0.01. We conclude that the regulation of HGL secretion by GLP-1 and CCK is altered in patients with gastritis. Moreover, GIP is a potent controller of HGL activity, both in healthy subjects and in patients with gastritis.

  18. Weapons of Mass Secretion : The Type III Secretion System of Shigella flexneri

    NARCIS (Netherlands)

    Sani, Musa

    2007-01-01

    In bacteria that are pathogenic for animals, type III secretion systems allow extracellular bacteria adhering to the surface of a host cell to inject specialized proteins across the plasma membrane. The injected proteins subvert the functioning of the aggressed cell or destroy its communications,

  19. Riemerella anatipestifer Type IX Secretion System Is Required for Virulence and Gelatinase Secretion

    Directory of Open Access Journals (Sweden)

    Yunqing Guo

    2017-12-01

    Full Text Available Riemerella anatipestifer (RA, a major causative agent of septicemia anserum exsudativa in domesticated ducklings, has a protein secretion system known as the type IX secretion system (T9SS. It is unknown whether the T9SS contributes to the virulence of RA through secretion of factors associated with pathogenesis. To answer this question, we constructed an RA mutant deficient in sprT, which encodes a core protein of the T9SS. Deletion of sprT yielded cells that failed to digest gelatin, an effect that was rescued via complementation by a plasmid encoding wild-type sprT. Complement-mediated killing was significantly increased in the deletion mutant, suggesting that proteins secreted by the T9SS are necessary for complement evasion in RA. Liquid chromatography-tandem mass spectrometry analysis revealed that RAYM_01812 and RAYM_04099 proteins containing a subtilisin-like serine protease domain and exhibiting extracellular gelatinase activity were secreted by the T9SS. Animal experiments demonstrated that the virulence of mutant strain ΔsprT strain was attenuated by 42,000-fold relative to wild-type RA-YM. Immunization with the ΔsprT protected ducks from challenge with RA-YM, suggesting that the former can be used as a live attenuated vaccine. These results indicate that the T9SS is functional in RA and contributes to its virulence by exporting key proteins. In addition, subtilisin-like serine proteases which are important virulence factors that interact with complement proteins may enable RA to evade immune surveillance in the avian innate immune system.

  20. Regulation of interleukin-6 secretion in murine pituicytes

    DEFF Research Database (Denmark)

    Thorn, Anders; Tuxen, Mikkel; Moesby, Lise

    2005-01-01

    Pituicytes, the astrocytic glial cells of the neural lobe, are known to secrete interleukin-6 and nitric oxide upon stimulation with various inflammatory mediators, i.e. interleukin-1beta. Nitric oxide is described to modulate the secretion of interleukin-6 in various cell types. The aim...... of the present study was to investigate the effect of nitric oxide on interleukin-1beta induced interleukin-6 secretion. Furthermore the effect of interferon-gamma on interleukin-6 and nitric oxide release was investigated. Cultures of pituicytes were prepared of neural lobes from male mice. The effect...... of interleukin-1beta and interferon-gamma on interleukin-6 and nitric oxide secretion was investigated in pituicytes cultured for 14 days. The secretion of interleukin-6 and nitric oxide was determined after 24 h of stimulation. Pituicytes secrete interleukin-6 upon stimulation with interleukin-1beta dose...

  1. Unconventional secretion of FABP4 by endosomes and secretory lysosomes.

    Science.gov (United States)

    Villeneuve, Julien; Bassaganyas, Laia; Lepreux, Sebastien; Chiritoiu, Marioara; Costet, Pierre; Ripoche, Jean; Malhotra, Vivek; Schekman, Randy

    2017-12-06

    An appreciation of the functional properties of the cytoplasmic fatty acid binding protein 4 (FABP4) has advanced with the recent demonstration that an extracellular form secreted by adipocytes regulates a wide range of physiological functions. Little, however, is known about the mechanisms that mediate the unconventional secretion of FABP4. Here, we demonstrate that FABP4 secretion is mediated by a membrane-bounded compartment, independent of the conventional endoplasmic reticulum-Golgi secretory pathway. We show that FABP4 secretion is also independent of GRASP proteins, autophagy, and multivesicular bodies but involves enclosure within endosomes and secretory lysosomes. We highlight the physiological significance of this pathway with the demonstration that an increase in plasma levels of FABP4 is inhibited by chloroquine treatment of mice. These findings chart the pathway of FABP4 secretion and provide a potential therapeutic means to control metabolic disorders associated with its dysregulated secretion. © 2018 Villeneuve et al.

  2. Combinatorial insulin secretion dynamics of recombinant hepatic and enteroendocrine cells.

    Science.gov (United States)

    Durvasula, Kiranmai; Thulé, Peter M; Sambanis, Athanassios

    2012-04-01

    One of the most promising cell-based therapies for combating insulin-dependent diabetes entails the use of genetically engineered non-β cells that secrete insulin in response to physiologic stimuli. A normal pancreatic β cell secretes insulin in a biphasic manner in response to glucose. The first phase is characterized by a transient stimulation of insulin to rapidly lower the blood glucose levels, which is followed by a second phase of insulin secretion to sustain the lowered blood glucose levels over a longer period of time. Previous studies have demonstrated hepatic and enteroendocrine cells to be appropriate hosts for recombinant insulin expression. Due to different insulin secretion kinetics from these cells, we hypothesized that a combination of the two cell types would mimic the biphasic insulin secretion of normal β cells with higher fidelity than either cell type alone. In this study, insulin secretion experiments were conducted with two hepatic cell lines (HepG2 and H4IIE) transduced with 1 of 3 adenoviruses expressing the insulin transgene and with a stably transfected recombinant intestinal cell line (GLUTag-INS). Insulin secretion was stimulated by exposing the cells to glucose only (hepatic cells), meat hydrolysate only (GLUTag-INS), or to a cocktail of the two secretagogues. It was found experimentally that the recombinant hepatic cells secreted insulin in a more sustained manner, whereas the recombinant intestinal cell line exhibited rapid insulin secretion kinetics upon stimulation. The insulin secretion profiles were computationally combined at different cell ratios to arrive at the combinatorial kinetics. Results indicate that combinations of these two cell types allow for tuning the first and second phase of insulin secretion better than either cell type alone. This work provides the basic framework in understanding the secretion kinetics of the combined system and advances it towards preclinical studies. Copyright © 2011 Wiley Periodicals

  3. A Modular Toolkit for Generating Pichia pastoris Secretion Libraries.

    Science.gov (United States)

    Obst, Ulrike; Lu, Timothy K; Sieber, Volker

    2017-06-16

    Yeasts are powerful eukaryotic hosts for the production of recombinant proteins due to their rapid growth to high cell densities and ease of genetic modification. For large-scale industrial production, secretion of a protein offers the advantage of simple and efficient downstream purification that avoids costly cell rupture, denaturation and refolding. The methylotrophic yeast Pichia pastoris (Komagataella phaffi) is a well-established expression host that has the ability to perform post-translational modifications and is generally regarded as safe (GRAS). Nevertheless, optimization of protein secretion in this host remains a challenge due to the multiple steps involved during secretion and a lack of genetic tools to tune this process. Here, we developed a toolkit of standardized regulatory elements specific for Pichia pastoris allowing the tuning of gene expression and choice of protein secretion tag. As protein secretion is a complex process, these parts are compatible with a hierarchical assembly method to enable the generation of large and diverse secretion libraries in order to explore a wide range of secretion constructs, achieve successful secretion, and better understand the regulatory factors of importance to specific proteins of interest. To assess the performance of these parts, we built and characterized the expression and secretion efficiency of 124 constructs that combined different regulatory elements with two fluorescent reporter proteins (RFP, yEGFP). Intracellular expression from our promoters was comparatively independent of whether RFP or yEGFP, and whether plasmid-based expression or genomically integrated expression, was used. In contrast, secretion efficiency significantly varied for different genes expressed using identical regulatory elements, with differences in secretion efficiency of >10-fold observed. These results highlight the importance of generating diverse secretion libraries when searching for optimal expression conditions, and

  4. DBSecSys: a database of Burkholderia mallei secretion systems

    OpenAIRE

    Memišević, Vesna; Kumar, Kamal; Cheng, Li; Zavaljevski, Nela; DeShazer, David; Wallqvist, Anders; Reifman, Jaques

    2014-01-01

    Background Bacterial pathogenicity represents a major public health concern worldwide. Secretion systems are a key component of bacterial pathogenicity, as they provide the means for bacterial proteins to penetrate host-cell membranes and insert themselves directly into the host cells’ cytosol. Burkholderia mallei is a Gram-negative bacterium that uses multiple secretion systems during its host infection life cycle. To date, the identities of secretion system proteins for B. mallei are not we...

  5. Regulation of stretch-activated ANP secretion by chloride channels.

    Science.gov (United States)

    Han, Jeong Hee; Bai, Guang Yi; Park, Jae-Hyeong; Yuan, Kuichang; Park, Woo Hyun; Kim, Sung Zoo; Kim, Suhn Hee

    2008-04-01

    This study was aimed to define roles of stretch-activated ion channels (SACs), especially Cl(-) channels, in regulation of atrial natriuretic peptide (ANP) secretion using isolated perfused beating atria. The volume load was achieved by elevating height of outflow catheter connected to isolated rat atria and the pressure load was achieved by decreasing diameter of outflow catheter. Both methods increased atrial contractility similarly although volume load was different (736microl for volume load vs. 129microl for pressure load). Atrial stretch by volume load markedly increased ECF translocation and ANP secretion but the pressure load slightly increased. The ANP secretion was positively correlated to workload generated by volume or pressure load. Treatment of atria with gadolinium, a blocker for SACs, attenuated the ECF translocation and the ANP secretion induced by volume load. A blocker for Ca2+-activated Cl(-) channel, niflumic acid (NFA), accentuated the ANP secretion induced by volume load whereas a blocker for swelling-activated Cl(-) channel, diisothiocyanatostilbene-2,2'-disulphonic acid (DIDS), attenuated the ANP secretion. The ANP secretion of hypertrophied atria by volume load was markedly reduced and the augmented effect of NFA on volume load-induced ANP secretion was not observed. These results indicate that Cl(-) channels may differently regulate stretch-activated ANP secretion.

  6. Security of the AES with a Secret S-Box

    DEFF Research Database (Denmark)

    Tiessen, Tyge; Knudsen, Lars Ramkilde; Kölbl, Stefan

    2015-01-01

    How does the security of the AES change when the S-box is replaced by a secret S-box, about which the adversary has no knowledge? Would it be safe to reduce the number of encryption rounds? In this paper, we demonstrate attacks based on integral cryptanalysis which allow to recover both the secret...... key and the secret S-box for respectively four, five, and six rounds of the AES. Despite the significantly larger amount of secret information which an adversary needs to recover, the attacks are very efficient with time/data complexities of 217/216, 238/240 and 290/264, respectively. Another...

  7. Indomethacin decreases gastroduodenal mucosal bicarbonate secretion in humans

    DEFF Research Database (Denmark)

    Mertz-Nielsen, A; Hillingsø, Jens; Bukhave, K

    1995-01-01

    BACKGROUND: Cyclooxygenase inhibitors reduce mucosal bicarbonate secretion in the duodenum, but the evidence for their effect on bicarbonate secretion in the stomach remains controversial. We have, therefore, studied how indomethacin influences gastroduodenal bicarbonate secretion and luminal...... release of prostaglandin (PG) E2 by means of a method that enables simultaneous measurements in the stomach and the duodenum. METHODS: Gastric and duodenal perfusions were performed twice in random order during control conditions or after pretreatment with indomethacin (100 mg intravenously) in eight...... basal and stimulated bicarbonate secretion, both in the stomach and in the duodenum....

  8. Pioglitazone acutely reduces energy metabolism and insulin secretion in rats

    National Research Council Canada - National Science Library

    Lamontagne, Julien; Jalbert-Arsenault, Elise; Pepin, Emilie; Peyot, Marie-Line; Ruderman, Neil B; Nolan, Christopher J; Joly, Erik; Madiraju, S R Murthy; Poitout, Vincent; Prentki, Marc

    2013-01-01

    Our objective was to determine if the insulin-sensitizing drug pioglitazone acutely reduces insulin secretion and causes metabolic deceleration in vivo independently of change in insulin sensitivity...

  9. Secret Sessions of Congress: A Brief Historical Overview

    National Research Council Canada - National Science Library

    Amer, Mildred

    2004-01-01

    Secret or closed door sessions of the House of Representatives and Senate are held periodically to discuss business, including impeachment deliberations, deemed to require confidentiality and secrecy...

  10. Secret Sessions of Congress: A Brief Historical Overview

    National Research Council Canada - National Science Library

    Amer, Mildred

    2005-01-01

    "Secret" or "closed door" sessions of the House of Representatives and Senate are held periodically to discuss business, including impeachment deliberations, deemed to require confidentiality and secrecy...

  11. Serotonergic involvement in stress-induced vasopressin and oxytocin secretion

    DEFF Research Database (Denmark)

    Jørgensen, Henrik; Knigge, Ulrich; Kjaer, Andreas

    2002-01-01

    OBJECTIVE: To investigate the involvement of serotonin (5-hydroxytryptamine - 5-HT) receptors in mediation of stress-induced arginine vasopressin (AVP) and oxytocin (OT) secretion in male rats. DESIGN: Experiments on laboratory rats with control groups. METHODS: Different stress paradigms were...... applied after pretreatment with intracerebroventricular infusion of saline or different 5-HT antagonists. RESULTS: Restraint stress (5 min), hypotensive hemorrhage or dehydration for 24 h increased AVP secretion fivefold and OT secretion threefold. Swim stress for 3 min had no effect on AVP secretion...

  12. Parathyroid hormone secretion in chronic renal failure

    DEFF Research Database (Denmark)

    Madsen, J C; Rasmussen, A Q; Ladefoged, S D

    1996-01-01

    The aim of study was to introduce and evaluate a method for quantifying the parathyroid hormone (PTH) secretion during hemodialysis in secondary hyperparathyroidism due to end-stage renal failure. We developed a method suitable for inducing sequential hypocalcemia and hypercalcemia during....../ionized calcium curves were constructed, and a mean calcium set-point of 1.16 mmol/liter was estimated compared to the normal mean of about 1.13 mmol/liter. In conclusion, we demonstrate that it is important to use a standardized method to evaluate parathyroid hormone dynamics in chronic renal failure. By the use...... of a standardized method we show that the calcium set-point is normal or slightly elevated, indicating normal parathyroid reactivity to calcium in chronic renal failure....

  13. Kinetic regulation of coated vesicle secretion

    CERN Document Server

    Foret, Lionel

    2008-01-01

    The secretion of vesicles for intracellular transport often rely on the aggregation of specialized membrane-bound proteins into a coat able to curve cell membranes. The nucleation and growth of a protein coat is a kinetic process that competes with the energy-consuming turnover of coat components between the membrane and the cytosol. We propose a generic kinetic description of coat assembly and the formation of coated vesicles, and discuss its implication to the dynamics of COP vesicles that traffic within the Golgi and with the Endoplasmic Reticulum. We show that stationary coats of fixed area emerge from the competition between coat growth and the recycling of coat components, in a fashion resembling the treadmilling of cytoskeletal filaments. We further show that the turnover of coat components allows for a highly sensitive switching mechanism between a quiescent and a vesicle producing membrane, upon a slowing down of the exchange kinetics. We claim that the existence of this switching behaviour, also tri...

  14. Multiple Ca2+ sensors in secretion

    DEFF Research Database (Denmark)

    Walter, Alexander M; Groffen, Alexander J; Sørensen, Jakob Balslev

    2011-01-01

    Regulated neurotransmitter secretion depends on Ca(2+) sensors, C2 domain proteins that associate with phospholipids and soluble N-ethylmaleimide-sensitive fusion attachment protein receptor (SNARE) complexes to trigger release upon Ca(2+) binding. Ca(2+) sensors are thought to prevent spontaneous...... fusion at rest (clamping) and to promote fusion upon Ca(2+) activation. At least eight, often coexpressed, Ca(2+) sensors have been identified in mammals. Accumulating evidence suggests that multiple Ca(2+) sensors interact, rather than work autonomously, to produce the complex secretory response...... observed in neurons and secretory cells. In this review, we present several working models to describe how different sensors might be arranged to mediate synchronous, asynchronous and spontaneous neurotransmitter release. We discuss the scenario that different Ca(2+) sensors typically act on one shared...

  15. Secrets of mlearning failures: confronting reality

    Directory of Open Access Journals (Sweden)

    Thomas Cochrane

    2012-08-01

    Full Text Available Having implemented and evaluated over 35 mlearning projects in a variety of contexts in higher education over the past 6 years the researcher is ready to share the untold secret: not all mlearning projects succeed! This article critiques three of the researcher's mlearning projects that can be classed as “failures” and compares them to successful projects to draw out critical implications for mlearning project design and implementation to avoid common pitfalls leading to potential project failure. This article uses the researcher's six critical success factors identified across the 35 mlearning projects to evaluate these three projects, and concludes that projects resulting in “no significant difference” in pedagogical outcomes are the result of attempts to shoehorn old pedagogies into new technologies. Lecturer professional development and sustained collaborative support are critical to fostering new pedagogies utilising the unique affordances of mobile devices.

  16. Secreted fungal aspartic proteases: A review.

    Science.gov (United States)

    Mandujano-González, Virginia; Villa-Tanaca, Lourdes; Anducho-Reyes, Miguel Angel; Mercado-Flores, Yuridia

    2016-01-01

    The aspartic proteases, also called aspartyl and aspartate proteases or acid proteases (E.C.3.4.23), belong to the endopeptidase family and are characterized by the conserved sequence Asp-Gly-Thr at the active site. These enzymes are found in a wide variety of microorganisms in which they perform important functions related to nutrition and pathogenesis. In addition, their high activity and stability at acid pH make them attractive for industrial application in the food industry; specifically, they are used as milk-coagulating agents in cheese production or serve to improve the taste of some foods. This review presents an analysis of the characteristics and properties of secreted microbial aspartic proteases and their potential for commercial application. Copyright © 2016 Asociación Española de Micología. Published by Elsevier Espana. All rights reserved.

  17. Efficient Secret Key Delivery Using Heartbeats

    Directory of Open Access Journals (Sweden)

    Cho Kwantae

    2017-01-01

    Full Text Available Recently many researchers have employed physiological signals like heartbeats as a source of the key seed used in key establishment protocols. The physiological signals make it easy to establish a secret key between implantable (or attachable medical devices which can sense physiological signals. A key establishment protocol is a fundamental requirement to support the security of the healthcare and medical services such as diagnosis, treatment, prevention, and follow-up services. However, existing key establishment protocols demand high computational and communication costs or need long key establishment time. In this paper, we propose an efficient IPI-based key establishment protocol that requires relatively short time while keeping the strength of security close.

  18. Parathyroid hormone secretion in chronic renal failure

    DEFF Research Database (Denmark)

    Madsen, J C; Rasmussen, A Q; Ladefoged, S D

    1996-01-01

    /ionized calcium curves were constructed, and a mean calcium set-point of 1.16 mmol/liter was estimated compared to the normal mean of about 1.13 mmol/liter. In conclusion, we demonstrate that it is important to use a standardized method to evaluate parathyroid hormone dynamics in chronic renal failure. By the use...... of a standardized method we show that the calcium set-point is normal or slightly elevated, indicating normal parathyroid reactivity to calcium in chronic renal failure.......The aim of study was to introduce and evaluate a method for quantifying the parathyroid hormone (PTH) secretion during hemodialysis in secondary hyperparathyroidism due to end-stage renal failure. We developed a method suitable for inducing sequential hypocalcemia and hypercalcemia during...

  19. A Novel Mechanism for Protein Delivery by the Type 3 Secretion System for Extracellularly Secreted Proteins.

    Science.gov (United States)

    Tejeda-Dominguez, Farid; Huerta-Cantillo, Jazmin; Chavez-Dueñas, Lucia; Navarro-Garcia, Fernando

    2017-03-28

    The type 3 secretion system (T3SS) is essential for bacterial virulence through delivering effector proteins directly into the host cytosol. Here, we identified an alternative delivery mechanism of virulence factors mediated by the T3SS, which consists of the association of extracellularly secreted proteins from bacteria with the T3SS to gain access to the host cytosol. Both EspC, a protein secreted as an enteropathogenic Escherichia coli (EPEC) autotransporter, and YopH, a protein detected on the surface of Yersinia, require a functional T3SS for host cell internalization; here we provide biophysical and molecular evidence to support the concept of the EspC translocation mechanism, which requires (i) an interaction between EspA and an EspC middle segment, (ii) an EspC translocation motif (21 residues that are shared with the YopH translocation motif), (iii) increases in the association and dissociation rates of EspC mediated by EspA interacting with EspD, and (iv) an interaction of EspC with the EspD/EspB translocon pore. Interestingly, this novel mechanism does not exclude the injection model (i.e., EspF) operating through the T3SS conduit; therefore, T3SS can be functioning as an internal conduit or as an external railway, which can be used to reach the translocator pore, and this mechanism appears to be conserved among different T3SS-dependent pathogens.IMPORTANCE The type 3 secretion system is essential for injection of virulence factors, which are delivered directly into the cytosol of the host cells for usurping and subverting host processes. Recent studies have shown that these effectors proteins indeed travel inside an "injectisome" conduit through a single step of translocation by connecting the bacterium and host cell cytoplasms. However, all findings are not compatible with this model. For example, both YopH, a protein detected on the surface of Yersinia, and EspC, an autotransporter protein secreted by enteropathogenic E. coli, require a functional T3

  20. An engineered yeast efficiently secreting penicillin.

    Directory of Open Access Journals (Sweden)

    Loknath Gidijala

    Full Text Available This study aimed at developing an alternative host for the production of penicillin (PEN. As yet, the industrial production of this beta-lactam antibiotic is confined to the filamentous fungus Penicillium chrysogenum. As such, the yeast Hansenula polymorpha, a recognized producer of pharmaceuticals, represents an attractive alternative. Introduction of the P. chrysogenum gene encoding the non-ribosomal peptide synthetase (NRPS delta-(L-alpha-aminoadipyl-L-cysteinyl-D-valine synthetase (ACVS in H. polymorpha, resulted in the production of active ACVS enzyme, when co-expressed with the Bacillus subtilis sfp gene encoding a phosphopantetheinyl transferase that activated ACVS. This represents the first example of the functional expression of a non-ribosomal peptide synthetase in yeast. Co-expression with the P. chrysogenum genes encoding the cytosolic enzyme isopenicillin N synthase as well as the two peroxisomal enzymes isopenicillin N acyl transferase (IAT and phenylacetyl CoA ligase (PCL resulted in production of biologically active PEN, which was efficiently secreted. The amount of secreted PEN was similar to that produced by the original P. chrysogenum NRRL1951 strain (approx. 1 mg/L. PEN production was decreased over two-fold in a yeast strain lacking peroxisomes, indicating that the peroxisomal localization of IAT and PCL is important for efficient PEN production. The breakthroughs of this work enable exploration of new yeast-based cell factories for the production of (novel beta-lactam antibiotics as well as other natural and semi-synthetic peptides (e.g. immunosuppressive and cytostatic agents, whose production involves NRPS's.

  1. Secretion of a pneumococcal type II secretion system pilus correlates with DNA uptake during transformation.

    Science.gov (United States)

    Balaban, Murat; Bättig, Patrick; Muschiol, Sandra; Tirier, Stephan M; Wartha, Florian; Normark, Staffan; Henriques-Normark, Birgitta

    2014-02-18

    Streptococcus pneumoniae is a major human pathogen that successfully adapts to the host environment via an efficient uptake system for free DNA liberated from other organisms in the upper respiratory tract, facilitating immune evasion and drug resistance. Although the initial signaling events leading to pneumococcal competence for DNA transformation and the fate of DNA when it has been taken up have been extensively studied, the actual mechanism by which DNA in the environment may traverse the thick capsular and cell wall layers remains unknown. Here we visualize that induction of competence results in the formation of a native morphologically distinct pilus structure on the bacterial surface. This plaited pilus is encoded by the competence (com)G locus, and, after assembly, it is rapidly released into the surrounding medium. Heterologous pneumococcal pilus expression in Escherichia coli was obtained by replacing the pulE-K putative pilin genes of the Klebsiella oxytoca type II secretion system with the complete comG locus. In the pneumococcus, the coordinated secretion of pili from the cells correlates to DNA transformation. A model for DNA transformation is proposed whereby pilus assembly "drills" a channel across the thick cell wall that becomes transiently open by secretion of the pilus, providing the entry port for exogenous DNA to gain access to DNA receptors associated with the cytoplasmic membrane.

  2. Macrophage-secreted factors inhibit ZAG expression and secretion by human adipocytes.

    Science.gov (United States)

    Gao, D; Trayhurn, P; Bing, C

    2010-08-30

    Zinc-alpha2-glycoprotein (ZAG), a novel adipokine, is downregulated in adipose tissue in obesity, a state characterized by increased adipose tissue macrophage infiltration and chronic low-grade inflammation. This study investigated whether macrophage-secreted factors and TNF-alpha, a major product of macrophages, modulate ZAG expression and secretion by human adipocytes. ZAG was produced primarily by adipocytes, and not by preadipocytes and macrophages. Incubation of preadipocytes with macrophage-conditioned medium for up to 12 days decreased ZAG mRNA and protein release, and the expression of adipogenic markers (PPARgamma and C/EBPalpha). Adipocytes treated with macrophage-conditioned medium for 24h displayed significant reductions in ZAG mRNA and release. Chronic TNF-alpha treatment let to significant decreases in ZAG expression and secretion, but marked upregulation of pro-inflammatory cytokines and chemokines (IL-6, leptin, IL-8, MCP-1 and RANTES) in adipocytes. These findings suggest that macrophage-associated inflammation may play a significant role in the downregulation of ZAG in adipose tissue in obesity. Copyright (c) 2010 Elsevier Ireland Ltd. All rights reserved.

  3. Bidrag til "Life begins at 100 - Secrets of the supercentenarians"

    DEFF Research Database (Denmark)

    von Bornemann Hjelmborg, Jacob

    2009-01-01

    Bidrag til: "Life begins at 100 - Secrets of the supercentenarians". Uddrag fra Human Genetics, vol. 119 p 1432. Udgivelsesdato: 5 september......Bidrag til: "Life begins at 100 - Secrets of the supercentenarians". Uddrag fra Human Genetics, vol. 119 p 1432. Udgivelsesdato: 5 september...

  4. Optimal Black-Box Secret Sharing over Arbitrary Abelian Groups

    DEFF Research Database (Denmark)

    Cramer, Ronald; Fehr, Serge

    2002-01-01

    A black-box secret sharing scheme for the threshold access structure T t,n is one which works over any finite Abelian group G. Briefly, such a scheme differs from an ordinary linear secret sharing scheme (over, say, a given finite field) in that distribution matrix and reconstruction vectors are ...

  5. Inhibition of secretion by asexual reproduction in Aspergillus niger

    NARCIS (Netherlands)

    Wang, Fengfeng|info:eu-repo/dai/nl/32304090X

    2014-01-01

    The aim of this Thesis was to study mechanism involved in zonal secretion in colonies of A. niger with emphasis on sporulation inhibited secretion. These mechanisms may be targets to improve A. nigeras a cell factory. I focused in particular on A. niger homologues of fluG and veA that are known

  6. In silico dissection of Type VII Secretion System components across ...

    Indian Academy of Sciences (India)

    2016-02-12

    Feb 12, 2016 ... [Das C, Ghosh TS and Mande SS 2016 In silico dissection of Type VII Secretion System components across bacteria: New directions towards functional characterization. J. Biosci. 41 133–143] DOI 10.1007/s12038-016-9599-8. 1. Introduction. Type VII Secretion System (T7SS) was first identified in.

  7. Hydrochlorothiazide increases interleukin-1 beta (IL-1β) secretion ...

    African Journals Online (AJOL)

    Previous studies showed that individuals with essential hypertension had increased interleukin-1beta (IL-1β) secretion by peripheral blood mononuclear cells (PBMCs) and also valsartan and simvastatin reduced this inflammatory marker. In this study, the effect of hydrochlorothiazide on IL-1β secretion by PBMCs in healthy ...

  8. Vote Markets, Latent Opportunism, and the Secret Ballot

    DEFF Research Database (Denmark)

    Justesen, Mogens Kamp; Bøttkjær, Louise Thorn; Gates, Scott

    The secret ballot is a cornerstone of modern democracy because it protects voter autonomy and allows voters to express their political preferences freely without fear of repercussions. In theory, the secret ballot is supposed to prevent vote buying – the exchange of votes for money or material...

  9. Oleanolic acid enhances insulin secretion in pancreatic beta-cells.

    NARCIS (Netherlands)

    Teodoro, T.; Zhang, L.; Alexander, T.; Yue, J.; Vranic, M.; Volchuk, A.

    2008-01-01

    We investigated the effect of oleanolic acid, a plant-derived triterpenoid, on insulin secretion and content in pancreatic beta-cells and rat islets. Oleanolic acid significantly enhanced insulin secretion at basal and stimulatory glucose concentrations in INS-1 832/13 cells and enhanced acute

  10. PACAP stimulates insulin secretion but inhibits insulin sensitivity in mice

    NARCIS (Netherlands)

    Filipsson, K; Pacini, G; Scheurink, AJW; Ahren, B

    Although pituitary adenylate cyclase-activating polypeptide (PACAP) stimulates insulin secretion, its net influence on glucose homeostasis in vivo has not been established. We therefore examined the action of PACAP-27 and PACAP-38 on insulin secretion, insulin sensitivity, and glucose disposal as

  11. Characterization of Pseudomonas aeruginosa Chitinase, a Gradually Secreted Protein

    NARCIS (Netherlands)

    Folders, J. (Jindra); Algra, J. (Jon); Roelofs, M.S. (Marc); Loon, L.C. van; Tommassen, J.P.M.; Bitter, Wilbert

    2001-01-01

    The gram-negative bacterium Pseudomonas aeruginosa secretes many proteins into its extracellular environment via the type I, II, and III secretion systems. In this study, a gene, chiC, coding for an extracellular chitinolytic enzyme, was identified. The chiC gene encodes a polypeptide of 483 amino

  12. Secretion of salivary statherin is compromised in uncontrolled diabetic patients

    Directory of Open Access Journals (Sweden)

    Masahiro Izumi

    2015-06-01

    Conclusions and general significance: The results show that synthesis and secretion of statherin is reduced in diabetics and this reduction is salivary gland specific. As compromised salivary statherin secretion leads to increased oral health risk, this study indicates that routine oral health assessment of these patients is warranted.

  13. The role of women's secret societies in cameroon's contemporary ...

    African Journals Online (AJOL)

    Although there are many secret societies, most of which belong to the male folk in the North West Province of Cameroon, little was and/or is known about their activities. However, Takumbeng, a women's secret society from the North West Province of Cameroon came to prominence in the 1990's during the political upheaval ...

  14. Antifungal activity of epithelial secretions from selected frog species ...

    African Journals Online (AJOL)

    This study aimed to investigate the antifungal activity of skin secretions from selected frogs (Amietia fuscigula, Strongylopus grayi and Xenopus laevis) and one toad (Amietophrynus pantherinus) of the south Western Cape Province of South Africa. Initially, different extraction techniques for the collection of skin secretions ...

  15. In silico dissection of Type VII Secretion System components across ...

    Indian Academy of Sciences (India)

    Type VII Secretion System (T7SS) is one of the factors involved in virulence of Mycobacteriun tuberculosis H37Rv. Numerous research efforts have been made in the last decade towards characterizing the components of this secretion system. An extensive genome-wide analysis through compilation of isolated information ...

  16. Renal renin secretion as regulator of body fluid homeostasis

    DEFF Research Database (Denmark)

    Damkjær, Mads; Isaksson, Gustaf L; Stubbe, Jane

    2013-01-01

    The renin-angiotensin system is essential for body fluid homeostasis and blood pressure regulation. This review focuses on the homeostatic regulation of the secretion of active renin in the kidney, primarily in humans. Under physiological conditions, renin secretion is determined mainly by sodium...

  17. Secretion of functional human interleukin-3 from Bacillus subtilis

    NARCIS (Netherlands)

    Westers, Lidia; Swaving Dijkstra, Dolf; Westers, Helga; van Dijl, Jan Maarten; Quax, Wim

    2006-01-01

    The Gram-positive bacterium Bacillus subtilis is well-known for its huge capacity to produce secreted bacterial enzymes. Nevertheless. the secretion of pharmaceutically interesting recombinant proteins by this organism is frequently inefficient. This paper documents for the first time on the

  18. Pathogenesis of canine cortisol-secreting adrenocortical tumors

    NARCIS (Netherlands)

    Kool, Miriam

    2015-01-01

    In dogs, hypercortisolism is one of the most frequently observed endocrine disorders, with an estimated incidence of about 1-2 cases per 1000 dogs per year. Approximately 15% of these cases is due to a cortisol-secreting adrenocortical tumor (AT). Cortisol-secreting ATs are characterized by

  19. Secrets and Misperceptions: The Creation of Self-Fulfilling Illusions

    Science.gov (United States)

    Cowan, Sarah K.

    2015-01-01

    This study examines who hears what secrets, comparing two similar secrets-one that is highly stigmatized and one that is less so. Using a unique survey representative of American adults and intake forms from a medical clinic, I document marked differences in who hears these secrets. People who are sympathetic to the stigmatizing secret are more likely to hear of it than those who may react negatively. This is a consequence of people not just selectively disclosing their own secrets but selectively sharing others’ as well. As a result, people in the same social network will be exposed to and influenced by different information about those they know and hence experience that network differently. When people effectively exist in networks tailored by others not to offend, then the information they hear tends to be that of which they already approve. Were they to hear secrets they disapproved of, then their attitudes might change, but they are less likely to hear those secrets. As such, the patterns of secret hearing contribute to a stasis in public opinion. PMID:26082932

  20. Sur les usages publics du secret : les archives du Service Secret Communiste Roumain

    Directory of Open Access Journals (Sweden)

    Andi Mihalache

    2009-10-01

    Full Text Available L’étude s’occupe des dossiers de l’ancienne police politique du régime communiste de Roumanie, la Securitate. Il a comme point de départ le fait que la mémoire sociale, le contexte politique et les débats publics actuels influencent la recherche historique, modifiant les perceptions sur le document d’archive. Ce dernier n’est plus vu seulement comme source principale de la recherche historique, devenant outil redoutable dans les disputes électorales, car, en base des informations extraites d’un dossier de la Securitate, l’on peut évincer un adversaire incommode. Se montrant utile à dénoncer la collaboration de quelqu’un sous le régime communiste, le document d’archive a acquis des mises symboliques imprévues, étant caché, protégé, volé, racheté, truqué, perdu et redécouvert à commande. Toutes ces pratiques compromettent de plus en plus les chances des historiens de reconstituer de manière crédible l’histoire récente des Roumains. On obtient difficilement l’accès aux dossiers de la Securitate, la patrimonialisation excessive des documents - en vérité leur trésorisation - étant conséquence directe du prestige idéologique et social que le secret détenait au temps communiste.This article focuses on the Securitate files as a political stake, but also as first rank sources for academic research, not from the current perspective of recent history, but from that of cultural history. In post-communist Romania, silence is the secret’s poor relative and confidentiality its respectable form. Ironically or not, we could state that in our country the secrets of communist repression are literally protected by law. The secret is an unbidden, illegitimate competence. In its ethics it does not matter what is good and what is bad, what is true and what is false. The main interest is what is said and what isn’t. The secret being a convention, a pact, the gesture of not saying seems to be more important than

  1. Proteomic Identification of Novel Secreted Antibacterial Toxins of the Serratia marcescens Type VI Secretion System*

    Science.gov (United States)

    Fritsch, Maximilian J.; Trunk, Katharina; Diniz, Juliana Alcoforado; Guo, Manman; Trost, Matthias; Coulthurst, Sarah J.

    2013-01-01

    It has recently become apparent that the Type VI secretion system (T6SS) is a complex macromolecular machine used by many bacterial species to inject effector proteins into eukaryotic or bacterial cells, with significant implications for virulence and interbacterial competition. “Antibacterial” T6SSs, such as the one elaborated by the opportunistic human pathogen, Serratia marcescens, confer on the secreting bacterium the ability to rapidly and efficiently kill rival bacteria. Identification of secreted substrates of the T6SS is critical to understanding its role and ability to kill other cells, but only a limited number of effectors have been reported so far. Here we report the successful use of label-free quantitative mass spectrometry to identify at least eleven substrates of the S. marcescens T6SS, including four novel effector proteins which are distinct from other T6SS-secreted proteins reported to date. These new effectors were confirmed as antibacterial toxins and self-protecting immunity proteins able to neutralize their cognate toxins were identified. The global secretomic study also unexpectedly revealed that protein phosphorylation-based post-translational regulation of the S. marcescens T6SS differs from that of the paradigm, H1-T6SS of Pseudomonas aeruginosa. Combined phosphoproteomic and genetic analyses demonstrated that conserved PpkA-dependent threonine phosphorylation of the T6SS structural component Fha is required for T6SS activation in S. marcescens and that the phosphatase PppA can reverse this modification. However, the signal and mechanism of PpkA activation is distinct from that observed previously and does not appear to require cell–cell contact. Hence this study has not only demonstrated that new and species-specific portfolios of antibacterial effectors are secreted by the T6SS, but also shown for the first time that PpkA-dependent post-translational regulation of the T6SS is tailored to fit the needs of different bacterial

  2. Omeprazole promotes proximal duodenal mucosal bicarbonate secretion in humans

    DEFF Research Database (Denmark)

    Mertz-Nielsen, Anette; Hillingsø, J; Bukhave, Klaus

    1996-01-01

    experiments. Also the combination of omeprazole and ranitidine increased (p=0.05) duodenal bicarbonate secretion, while ranitidine alone caused no change in either basal or stimulated secretion. In the stomach basal as well as vagally stimulated bicarbonate secretion was independent of the means of acid......The proton pump inhibitor, omeprazole, surprisingly resulted in higher rates of proximal duodenal mucosal bicarbonate secretion than previously reported using an H-2 receptor antagonist for gastric acid inhibition. Gastroduodenal perfusions were performed in healthy volunteers to evaluate whether...... this incidental finding is explained by more potent gastric acid inhibition by omeprazole or might be caused by the different mode of drug action. Basal and stimulated gastric and duodenal bicarbonate secretion rates were measured in the same subjects in control experiments (n=17) and after pretreatment with high...

  3. When confidentiality mandates a secret be kept: a case report.

    Science.gov (United States)

    Hough, G

    1992-01-01

    The success of group psychotherapy is impeded when group members maintain secrets and fail to self-disclose to other group members. However, revealing secrets sometimes conflicts with the legal and ethical constraints of confidentiality, thus placing a greater burden on the group. The case reported here illustrates how complex the dynamics of group therapy become when one group member maintains a secret. In this case, one member maintained a secret about another to preserve that individual's right to privacy and confidentiality. This case also details the group member's deliberations and confusion about the effect his secret had on the group's success, as well as the burden he experienced by choosing to preserve confidentiality at the expense of successful therapy for himself.

  4. Unconventional Secretion of Heat Shock Proteins in Cancer

    Science.gov (United States)

    Santos, Tiago Góss; Martins, Vilma Regina; Hajj, Glaucia Noeli Maroso

    2017-01-01

    Heat shock proteins (HSPs) are abundant cellular proteins involved with protein homeostasis. They have both constitutive and inducible isoforms, whose expression levels are further increased by stress conditions, such as temperature elevation, reduced oxygen levels, infection, inflammation and exposure to toxic substances. In these situations, HSPs exert a pivotal role in offering protection, preventing cell death and promoting cell recovery. Although the majority of HSPs functions are exerted in the cytoplasm and organelles, several lines of evidence reveal that HSPs are able to induce cell responses in the extracellular milieu. HSPs do not possess secretion signal peptides, and their secretion was subject to widespread skepticism until the demonstration of the role of unconventional secretion forms such as exosomes. Secretion of HSPs may confer immune system modulation and be a cell-to-cell mediated form of increasing stress resistance. Thus, there is a wide potential for secreted HSPs in resistance of cancer therapy and in the development new therapeutic strategies. PMID:28468249

  5. The Shaggy Inc Cap medicinal mushroom, Coprinus comatus (O.F.Mull.: Fr.) Pers. (Agaricomycetideae) substances interfere with H2O2 induction of the NF-kappaB pathway through inhibition of Ikappaalpha phosphorylation in MCF7 breast cancer cells.

    Science.gov (United States)

    Asatiani, Mikheil D; Wasser, Solomon P; Nevo, Eviatar; Ruimi, Nili; Mahajna, Jamal; Reznick, Abraham Z

    2011-01-01

    Breast cancer is the most commonly diagnosed cancer among women. Currently, there is no effective therapy for malignant estrogen-independent breast cancer. In our study, we used hydrogen peroxide, a well-known strong oxidative reagent capable of activating the nuclear factor kappa B (NF-kappaB) transcription factor. The IC50 value of the culinary-medicinal Shaggy Inc Cap mushroom Coprinus comatus culture liquid crude extract on MCF7 cell viability was found to be as low as 76 microg/mL, and the IC50 value of C. comatus ethyl acetate extract was only 32 microg/ mL. Our results also showed that both extracts significantly affected IkappaBalpha phosphorylation in a dose-dependent manner. The effect of ethyl acetate extract was comparable to the effect of curcumin, a known NF-kappaB pathway inhibitor, and seemed to be the most active inhibitor of H2O2-dependent IkappaBalpha phosphorylation. In addition, the data obtained showed that only ethyl acetate extract inhibited the activity of IKK complex, at close to 90% as compared to the control of the untreated sample. These results suggest that C. comatus contains potent compounds capable of inhibiting NF-kappaB function and also possibly acts as an antitumor agent.

  6. The Haloprotease CPI Produced by the Moderately Halophilic Bacterium Pseudoalteromonas ruthenica Is Secreted by the Type II Secretion Pathway▿ †

    Science.gov (United States)

    Sánchez-Porro, Cristina; Mellado, Encarnación; Pugsley, Anthony P.; Francetic, Olivera; Ventosa, Antonio

    2009-01-01

    The gene (cpo) encoding the extracellular protease CPI produced by the moderately halophilic bacterium Pseudoalteromonas ruthenica CP76 was cloned, and its nucleotide sequence was analyzed. The cpo gene encodes a 733-residue protein showing sequence similarity to metalloproteases of the M4 family. The type II secretion apparatus was shown to be responsible for secretion of the haloprotease CPI. PMID:19376897

  7. Identification of genes encoding the type IX secretion system and secreted proteins in Flavobacterium columnare IA-S-4

    Science.gov (United States)

    Flavobacterium columnare, a member of the phylum Bacteroidetes, causes columnaris disease in wild and aquaculture-reared freshwater fish. The mechanisms responsible for columnaris disease are not known. Many members of the phylum Bacteroidetes use type IX secretion systems (T9SSs) to secrete enzymes...

  8. "ON-AIR SECRETS": TEN YEARS LATER

    Directory of Open Access Journals (Sweden)

    Bysko Maxim V.

    2013-12-01

    Full Text Available The article is a scientific review on the training manual On-air secret by Andrey Bubukin, published in 2003. This guide has been written by a man who never had anything to do with radio, but by the will of fate suddenly became at once the program director of Russia's first non-state radio station Europa Plus Moscow, the commercial music radio. Then came to the fore not the questions of art qualities of broadcast, but technical-mathematical details of the automated programming of song material. Therefore, at the end of the Soviet broadcasting, it became possible appearance some people obviously alien to the electronic mass media. They are, however, strongly influenced the further "format" broadcasting in new Russia, so as the principles of songs rotation and playlist making on the radio still can’t do without recourse to the Bubukin's textbook. There are serious concerns about the fact that the book is intended not only for program directors, but for the music editors also. Extremely dubious assertions and recommendations related to the problems of arts, journalism, radio-presenter's skill, history of radio.

  9. Secrets of the old one Einstein, 1905

    CERN Document Server

    Bernstein, Jeremy

    2006-01-01

    In March 1905, at approximately eight week intervals, the Editor of the noted German physics journal, Annalen der Physik, received three hand-written manuscripts from a relatively unknown patent examiner in Bern, Switzerland. This patent examiner was the twenty-six year old Albert Einstein and the three papers would set the agenda for twentieth century physics. A fourth short paper was received in September 1905 and contained Einstein's derivation of the formula E=mc^2. These papers changed our lives in the twentieth century and beyond. While to a professional physicist the mathematics in these papers are quite straight forward, the ideas behind them are not. In fact, none of Einstein's contemporaries fully understood what he had done. In SECRETS OF THE OLD ONE: Einstein, 1905, renowned science writer Jeremy Bernstein makes these ideas accessible to a general reader with a limited background in mathematics. After reading this book, you will understand why 1905 is often designated as Einstein's miracle year.

  10. Undocumented secrets of MATLAB-Java programming

    CERN Document Server

    Altman, Yair

    2012-01-01

    For a variety of reasons, the MATLAB®-Java interface was never fully documented. This is really quite unfortunate: Java is one of the most widely used programming languages, having many times the number of programmers and programming resources as MATLAB. Also unfortunate is the popular claim that while MATLAB is a fine programming platform for prototyping, it is not suitable for real-world, modern-looking applications. Undocumented Secrets of MATLAB®-Java Programming aims to correct this misconception. This book shows how using Java can significantly improve MATLAB program appearance and functionality, and that this can be done easily and even without any prior Java knowledge. Readers are led step-by-step from simple to complex customizations. Code snippets, screenshots, and numerous online references are provided to enable the utilization of this book as both a sequential tutorial and as a random-access reference suited for immediate use. Java-savvy readers will find it easy to tailor code samples for th...

  11. Type I signal peptidase and protein secretion in Staphylococcus epidermidis.

    Science.gov (United States)

    Powers, Michael E; Smith, Peter A; Roberts, Tucker C; Fowler, Bruce J; King, Charles C; Trauger, Sunia A; Siuzdak, Gary; Romesberg, Floyd E

    2011-01-01

    Bacterial protein secretion is a highly orchestrated process that is essential for infection and virulence. Despite extensive efforts to predict or experimentally detect proteins that are secreted, the characterization of the bacterial secretome has remained challenging. A central event in protein secretion is the type I signal peptidase (SPase)-mediated cleavage of the N-terminal signal peptide that targets a protein for secretion via the general secretory pathway, and the arylomycins are a class of natural products that inhibit SPase, suggesting that they may be useful chemical biology tools for characterizing the secretome. Here, using an arylomycin derivative, along with two-dimensional gel electrophoresis and liquid chromatography-tandem mass spectrometry (LC-MS/MS), we identify 11 proteins whose secretion from stationary-phase Staphylococcus epidermidis is dependent on SPase activity, 9 of which are predicted to be translated with canonical N-terminal signal peptides. In addition, we find that the presence of extracellular domains of lipoteichoic acid synthase (LtaS) and the β-lactam response sensor BlaR1 in the medium is dependent on SPase activity, suggesting that they are cleaved at noncanonical sites within the protein. In all, the data define the proteins whose stationary-phase secretion depends on SPase and also suggest that the arylomycins should be valuable chemical biology tools for the study of protein secretion in a wide variety of different bacteria.

  12. Secretion of Streptomyces tendae antifungal protein 1 by Lactococcus lactis

    Directory of Open Access Journals (Sweden)

    D.A. Freitas

    2005-11-01

    Full Text Available Lactococcus lactis, the model lactic acid bacterium, is a good candidate for heterologous protein production in both foodstuffs and the digestive tract. We attempted to produce Streptomyces tendae antifungal protein 1 (Afp1 in L. lactis with the objective of constructing a strain able to limit fungal growth. Since Afp1 activity requires disulfide bond (DSB formation and since intracellular redox conditions are reportedly unfavorable for DSB formation in prokaryotes, Afp1 was produced as a secreted form. An inducible expression-secretion system was used to drive Afp1 secretion by L. lactis; Afp1 was fused or not with LEISSTCDA, a synthetic propeptide (LEISS that has been described to be a secretion enhancer. Production of Afp1 alone was not achieved, but production of LEISS-Afp1 was confirmed by Western blot and immunodetection with anti-Afp1 antibodies. This protein (molecular mass: 9.8 kDa is the smallest non-bacteriocin heterologous protein ever reported to be secreted in L. lactis via the Sec-dependent pathway. However, no anti-fungal activity was detected, even in concentrated samples of induced supernatant. This could be due to a too low secretion yield of Afp1 in L. lactis, to the absence of DSB formation, or to an improper DSB formation involving the additional cysteine residue included in LEISS propeptide. This raises questions about size limits, conformation problems, and protein secretion yields in L. lactis.

  13. Real-time single-cell imaging of protein secretion

    Science.gov (United States)

    Shirasaki, Yoshitaka; Yamagishi, Mai; Suzuki, Nobutake; Izawa, Kazushi; Nakahara, Asahi; Mizuno, Jun; Shoji, Shuichi; Heike, Toshio; Harada, Yoshie; Nishikomori, Ryuta; Ohara, Osamu

    2014-01-01

    Protein secretion, a key intercellular event for transducing cellular signals, is thought to be strictly regulated. However, secretion dynamics at the single-cell level have not yet been clarified because intercellular heterogeneity results in an averaging response from the bulk cell population. To address this issue, we developed a novel assay platform for real-time imaging of protein secretion at single-cell resolution by a sandwich immunoassay monitored by total internal reflection microscopy in sub-nanolitre-sized microwell arrays. Real-time secretion imaging on the platform at 1-min time intervals allowed successful detection of the heterogeneous onset time of nonclassical IL-1β secretion from monocytes after external stimulation. The platform also helped in elucidating the chronological relationship between loss of membrane integrity and IL-1β secretion. The study results indicate that this unique monitoring platform will serve as a new and powerful tool for analysing protein secretion dynamics with simultaneous monitoring of intracellular events by live-cell imaging. PMID:24751898

  14. Nasal secretion of the ozone scavenger uric acid

    Energy Technology Data Exchange (ETDEWEB)

    Peden, D.B.; Swiersz, M.; Ohkubo, K.; Hahn, B.; Emery, B.; Kaliner, M.A. (Univ. of North Carolina, Chapel Hill (United States))

    1993-08-01

    Uric acid, an important scavenger of ozone, has been identified as the major low molecular weight antioxidant in baseline and cholinergically induced nasal secretions. The purpose of this study was to determine the specific tissue source of uric acid in airway secretions. The secretion of uric acid is increased by cholinergic stimulation and correlates closely with the secretion of lactoferrin (a nasal glandular protein), suggesting that submucosal glands are involved. Indeed, nasal turbinate tissue was found to contain uric acid. However, careful analysis of nasal turbinate tissue failed to reveal the presence of xanthine oxidase, the enzyme responsible for uric acid synthesis. These data suggest that uric acid might be taken up secondarily by glands from plasma. This possibility was strengthened by the observation that lowering the plasma urate level with probenecid concomitantly lowered urate secretion. These findings are consistent with the hypotheses that the principal source of uric acid in nasal secretions is plasma and that uric acid is taken up, concentrated, and secreted by nasal glands.

  15. Stories and Dreams, Memories and Secrets : Functions of Narration in Amy Tan's The Hundred Secret Senses

    OpenAIRE

    Niblaeus, Frida

    2013-01-01

    The aim of this dissertation is to explore the functions of narration in Amy Tan’s novel The Hundred Secret Senses. The dissertation is divided into three parts: 1 ‘Introduction’, 2 ‘Analysis’ and 3 ‘Conclusion’. After presenting the writings of Amy Tan and my chosen primary literature, I give a brief survey of terms, theories and previous research. Part 2 ‘Analysis’ is presented in an order that corresponds approximately to the chronology of the primary literature, and will be divided into t...

  16. Activated platelets enhance IL-10 secretion and reduce TNF-α secretion by monocytes

    DEFF Research Database (Denmark)

    Gudbrandsdottir, Sif; Hasselbalch, Hans C; Nielsen, Claus H

    2013-01-01

    ), Escherichia coli LPS, or intact Porphyromonas gingivalis. Addition of platelets activated by thrombin-receptor-activating peptide enhanced IL-10 production induced by LPS (p ....05), and P. gingivalis (p IL-10 and TNF-α production were observed on addition of platelet supernatant to mononuclear cells, whereas addition of recombinant soluble CD40L mimicked the effects on IL-10...... production. Moreover, Ab-mediated blockade of CD40L counteracted the effect of platelets and platelet supernatants on TNF-α production. Monocytes separated into two populations with respect to IL-10 production induced by TG; the high-secreting fraction increased from 0.8 to 2.1% (p

  17. Neurogenic mucosal bicarbonate secretion in guinea pig duodenum.

    Science.gov (United States)

    Fei, G; Fang, X; Wang, G D; Liu, S; Wang, X Y; Xia, Y; Wood, J D

    2013-02-01

    To test a hypothesis that: (i) duodenal pH and osmolarity are individually controlled at constant set points by negative feedback control centred in the enteric nervous system (ENS); (ii) the purinergic P2Y(1) receptor subtype is expressed by non-cholinergic secretomotor/vasodilator neurons, which represent the final common excitatory pathway from the ENS to the bicarbonate secretory glands. Ussing chamber and pH-stat methods investigated involvement of the P2Y(1) receptor in neurogenic stimulation of mucosal bicarbonate (HCO(3)(-)) secretion in guinea pig duodenum. ATP increased HCO(3)(-) secretion with an EC(50) of 160 nM. MRS2179, a selective P2Y(1) purinergic receptor antagonist, suppressed ATP-evoked HCO(3)(-) secretion by 47% and Cl(-) secretion by 63%. Enteric neuronal blockade by tetrodotoxin or exposure to a selective vasoactive intestinal peptide (VIP, VPAC(1)) receptor antagonist suppressed ATP-evoked HCO(3)(-) secretion by 61 and 41%, respectively, and Cl- by 97 and 70% respectively. Pretreatment with the muscarinic antagonist, scopolamine did not alter ATP-evoked HCO3(-) or Cl(-) secretion. Whereas acid directly stimulates the mucosa to release ATP and stimulate HCO(3)(-) secretion in a cytoprotective manner, neurogenically evoked HCO(3)(-) secretion accounts for feedback control of optimal luminal pH for digestion. ATP stimulates duodenal HCO(3)(-) secretion through an excitatory action at purinergic P2Y(1) receptors on neurons in the submucosal division of the ENS. Stimulation of the VIPergic non-cholinergic secretomotor/vasodilator neurons, which are one of three classes of secretomotor neurons, accounts for most, if not all, of the neurogenic secretory response evoked by ATP. © 2012 The Authors. British Journal of Pharmacology © 2012 The British Pharmacological Society.

  18. A secreted factor represses cell proliferation in Dictyostelium

    OpenAIRE

    Brock, Debra A.; Gomer, Richard H.

    2005-01-01

    Many cells appear to secrete factors called chalones that limit their proliferation, but in most cases the factors have not been identified. We found that growing Dictyostelium cells secrete a 60 kDa protein called AprA for autocrine proliferation repressor. AprA has similarity to putative bacterial proteins of unknown function. Compared with wild-type cells, aprA-null cells proliferate faster, while AprA overexpressing cells proliferate slower. Growing wild-type cells secrete a factor that i...

  19. Regulation of gut hormone secretion. Studies using isolated perfused intestines

    DEFF Research Database (Denmark)

    Svendsen, Berit; Holst, Jens Juul.

    2016-01-01

    hormones is highly increased after gastric bypass operations, which have turned out to be an effective therapy of not only obesity but also type 2 diabetes. These effects are likely to be due, at least in part, to increases in the secretion of these gut hormones (except GIP). Therefore, stimulation...... of the endogenous hormone represents an appealing therapeutic strategy, which has spurred an interest in understanding the regulation of gut hormone secretion and a search for particularly GLP-1 and PYY secretagogues. The secretion of the gut hormones is stimulated by oral intake of nutrients often including...

  20. Insulin secretion and action in North Indian women during pregnancy

    DEFF Research Database (Denmark)

    Arora, G P; Almgren, P; Thaman, R G

    2017-01-01

    with normal glucose tolerance, and furthermore displayed lower insulin secretion than GDM1999 women. Urban habitat, illiteracy, high age and low BMI were independently associated with reduced insulin secretion, whereas Sikh religion, increasing age and BMI, as well as a family history of diabetes were....... RESULTS: Gestational diabetes defined using both criteria was associated with decreased insulin secretion compared with pregnant women with normal glucose tolerance. Women with gestational diabetes defined by the adapted GDM2013, but not GDM1999 criteria, were more insulin resistant than pregnant women...

  1. Towards Shift Tolerant Visual Secret Sharing Schemes without Pixel Expansion

    Directory of Open Access Journals (Sweden)

    Juan Justie Su-Tzu

    2016-01-01

    Full Text Available Naor and Shamir proposed the visual cryptography in 1995, they encrypted secret image into two meaningless random images, called shares, and it can be decrypted by human vision without any calculations. However, there would be problems in alignment when these two shares are staked by hand in practical. Therefore, this paper presents the fault-tolerant schemes of stacking two shares which are acquired from secret image encryption without pixel expansion. The main idea of these schemes is combining several pixels to be a unit, then encrypting every unit into a specific combination of pixels. It makes visual secret sharing scheme more practical.

  2. Cryptanalysis and improvement of multiparty quantum secret sharing schemes

    Energy Technology Data Exchange (ETDEWEB)

    Wang Tianyin [State key Laboratory of Networking and Switching Technology, Beijing University of Posts and Telecommunications, Beijing 100876 (China); School of Mathematical Science, Luoyang Normal University, Luoyang 471022 (China)], E-mail: wangtianyin@yahoo.cn; Wen Qiaoyan; Gao Fei; Lin Song [State key Laboratory of Networking and Switching Technology, Beijing University of Posts and Telecommunications, Beijing 100876 (China); Zhu Fuchen [National Laboratory for Modern Communications, PO Box 810, Chengdu 610041 (China)

    2008-12-22

    A scheme of multiparty quantum secret sharing of classical messages (QSSCM) [Z.J. Zhang et al., Opt. Commun. 269 (2007) 418] was proposed. Lin et al. [S. Lin et al., Opt. Commun. 281 (2008) 4553] showed the last agent can obtain half of the secret in Z.J. Zhang's et al. three-party QSSCM scheme and gave an improved version. We further show the first agent and the last agent can obtain all the secret without introducing any error in Zhang's et al. multiparty QSSCM scheme by a special attack with quantum teleportation. We also present an improved version.

  3. Cryptanalysis of quantum secret sharing based on GHZ states

    Energy Technology Data Exchange (ETDEWEB)

    Liu Xiaofen; Pan Rijing, E-mail: liuxiaofen23@fjnu.edu.cn [School of Mathematics and Computer Science, Fujian Normal University, Fuzhou 350007 (China); Key Laboratory of Network Security and Cryptography, Fujian Normal University, Fuzhou 350007 (China)

    2011-10-15

    In a recent paper (Hwang et al 2011 Phys. Scr. 83 045004), a subtle quantum secret sharing (QSS) protocol was proposed. One major advantage of this protocol is its high efficiency; that is, it is more efficient than other QSS protocols using the Greenberger-Horne-Zeilinger states. However, we analyzed the security of this protocol and found that it is insecure for one agent who may deduce half of the information about the deal's secret directly just by his/her piece of secret. Finally, a feasible improvement of this QSS protocol is put forward.

  4. Secret letters cast light on Copenhagen

    Energy Technology Data Exchange (ETDEWEB)

    Durrani, Matin

    2001-11-01

    Letters by Niels Bohr that have been kept secret since his death could explain the mystery of why Werner Heisenberg visited him in Copenhagen in 1941. When the author Michael Frayn spent two years writing Copenhagen, he had no idea how successful the play would become. He doubted that audiences would sit through a historical drama about a war-time meeting between Werner Heisenberg - head of Germany's nuclear programme - and his old mentor Niels Bohr in the Nazi-occupied Danish capital in 1941. But Frayn's efforts paid off. Audiences and critics alike have thrilled at the way the award-winning play probes the historical uncertainty that surrounds the encounter. Was Heisenberg fishing for information about the Allies' atomic plans - or was he trying to recruit Bohr for Germany's bomb programme? Did Heisenberg want to suggest that the Germans were close to finishing a bomb so that the Allies would make peace with Hitler? Maybe he was simply seeking approval from Bohr for his own atomic work. There is also a moral debate: did Heisenberg know how to build a bomb, but decided not to - or did he want to build one, but got his calculations wrong? Unfortunately, no-one was there to record or observe the encounter and we cannot know for sure what was said or implied between the two men. All we do know is that the pair dined together and took a short walk - and that the incident damaged Bohr and Heisenberg's friendship forever. To piece together what happened, historians of science have had to rely on Heisenberg's post-war recollections - which have been ambiguous and contradictory - along with scraps of evidence from secondary sources. (U.K.)

  5. Burkholderia cenocepacia type VI secretion system mediates escape of type II secreted proteins into the cytoplasm of infected macrophages.

    Directory of Open Access Journals (Sweden)

    Roberto Rosales-Reyes

    Full Text Available Burkholderia cenocepacia is an opportunistic pathogen that survives intracellularly in macrophages and causes serious respiratory infections in patients with cystic fibrosis. We have previously shown that bacterial survival occurs in bacteria-containing membrane vacuoles (BcCVs resembling arrested autophagosomes. Intracellular bacteria stimulate IL-1β secretion in a caspase-1-dependent manner and induce dramatic changes to the actin cytoskeleton and the assembly of the NADPH oxidase complex onto the BcCV membrane. A Type 6 secretion system (T6SS is required for these phenotypes but surprisingly it is not required for the maturation arrest of the BcCV. Here, we show that macrophages infected with B. cenocepacia employ the NLRP3 inflammasome to induce IL-1β secretion and pyroptosis. Moreover, IL-1β secretion by B. cenocepacia-infected macrophages is suppressed in deletion mutants unable to produce functional Type VI, Type IV, and Type 2 secretion systems (SS. We provide evidence that the T6SS mediates the disruption of the BcCV membrane, which allows the escape of proteins secreted by the T2SS into the macrophage cytoplasm. This was demonstrated by the activity of fusion derivatives of the T2SS-secreted metalloproteases ZmpA and ZmpB with adenylcyclase. Supporting this notion, ZmpA and ZmpB are required for efficient IL-1β secretion in a T6SS dependent manner. ZmpA and ZmpB are also required for the maturation arrest of the BcCVs and bacterial intra-macrophage survival in a T6SS-independent fashion. Our results uncover a novel mechanism for inflammasome activation that involves cooperation between two bacterial secretory pathways, and an unanticipated role for T2SS-secreted proteins in intracellular bacterial survival.

  6. ACTH-secreting bronchial carcinoid: a diagnostic and therapeutic challenge.

    LENUS (Irish Health Repository)

    McDermott, J H

    2008-09-01

    We describe a case of Cushing\\'s syndrome due to ectopic ACTH secretion, where the only potential source on conventional imaging was a tiny benign-appearing lung nodule, which failed to take up radiolabelled octreotide.

  7. Secrets of universe eyed as collider core installed

    CERN Document Server

    Evans, Robert

    2007-01-01

    "The world's leading centre for research into the origins of matter on Wedneday took a giant step towards completion of 1 15-year project which scientists hope will unlock many secrets of the universe."(1 page)

  8. Secrets of universe eyed as collider core installed

    CERN Document Server

    Evans, Robert

    2007-01-01

    "The world's leading centre for researach into the origins of matter on Wednesday took a giant step towards completion of a 15-year project which scientists hope will unlock many secrets of the universe." (1/2 page)

  9. Ghrelin drives GH secretion during fasting in man

    NARCIS (Netherlands)

    A.F. Muller (Alex); S.W.J. Lamberts (Steven); L.J. Hofland (Leo); M. Bidlingmaier (Martin); C.J. Strasburger; E. Ghigo (Ezio); A-J. van der Lely (Aart-Jan); J.A.M.J.L. Janssen (Joseph); P.M. van Koetsveld (Peter)

    2002-01-01

    textabstractOBJECTIVES: In humans, fasting leads to elevated serum GH concentrations. Traditionally, changes in hypothalamic GH-releasing hormone and somatostatin release are considered as the main mechanisms that induce this elevated GH secretion during fasting. Ghrelin is an

  10. [Effect of the antihypoxic agent isothiobarbamine on gastric secretion].

    Science.gov (United States)

    Troitskaia, V B; Repetun, I N; Poliakova, M A

    1986-01-01

    The experiments on dogs with the isolated Pavlov's ventricles showed that isothiobarbamine (30 mg/kg) in the days of administration decreased the volume of the gastric juice secretion and its total and free activity.

  11. Reappraisal of bicarbonate secretion by the human oesophagus

    DEFF Research Database (Denmark)

    Mertz-Nielsen, A; Hillingsø, J; Bukhave, Klaus

    1997-01-01

    BACKGROUND AND AIMS: Administration of omeprazole to healthy volunteers was recently reported to increase proximal duodenal mucosalbicarbonate secretion. As human oesophagus also secretes bicarbonate, the hypothesis was tested that omeprazole may stimulate oesophagealbicarbonate secretion and thus....../day omeprazole for three days and 80 mg intravenous omeprazole before perfusionor 600 mg/day ranitidine for three days and 50 mg/h intravenously during the perfusion. Saliva and samples of aspirate from the perfusedoesophagus and stomach were collected and bicarbonate concentrations were measured. RESULTS......: The median rates (95% confidence intervals)of intrinsic oesophageal bicarbonate secretion, corrected for contaminating salivary and gastric bicarbonate, were 89 (33-150) and 121 (63-203)mumol/h/10 cm (p > 0.5) in omeprazole and ranitidine treated subjects respectively. Salivary and gastric bicarbonate...

  12. Reappraisal of bicarbonate secretion by the human oesophagus

    DEFF Research Database (Denmark)

    Mertz-Nielsen, A; Hillingsø, Jens; Bukhave, K

    1997-01-01

    BACKGROUND AND AIMS: Administration of omeprazole to healthy volunteers was recently reported to increase proximal duodenal mucosal bicarbonate secretion. As human oesophagus also secretes bicarbonate, the hypothesis was tested that omeprazole may stimulate oesophageal bicarbonate secretion...... with either 60 mg/day omeprazole for three days and 80 mg intravenous omeprazole before perfusion or 600 mg/day ranitidine for three days and 50 mg/h intravenously during the perfusion. Saliva and samples of aspirate from the perfused oesophagus and stomach were collected and bicarbonate concentrations were...... measured. RESULTS: The median rates (95% confidence intervals) of intrinsic oesophageal bicarbonate secretion, corrected for contaminating salivary and gastric bicarbonate, were 89 (33-150) and 121 (63-203) mumol/h/10 cm (p > 0.5) in omeprazole and ranitidine treated subjects respectively. Salivary...

  13. High-yield secretion of multiple client proteins in Aspergillus.

    Science.gov (United States)

    Segato, Fernando; Damásio, André R L; Gonçalves, Thiago A; de Lucas, Rosymar C; Squina, Fabio M; Decker, Stephen R; Prade, Rolf A

    2012-07-15

    Production of pure and high-yield client proteins is an important technology that addresses the need for industrial applications of enzymes as well as scientific experiments in protein chemistry and crystallization. Fungi are utilized in industrial protein production because of their ability to secrete large quantities of proteins. In this study, we engineered a high-expression-secretion vector, pEXPYR that directs proteins towards the extracellular medium in two Aspergillii host strains, examine the effect of maltose-induced over-expression and protein secretion as well as time and pH-dependent protein stability in the medium. We describe five client proteins representing a core set of hemicellulose degrading enzymes that accumulated up to 50-100 mg/L of protein. Using a recyclable genetic marker that allows serial insertion of multiple genes, simultaneous hyper-secretion of three client proteins in a single host strain was accomplished. Copyright © 2012 Elsevier Inc. All rights reserved.

  14. The secretory synapse: the secrets of a serial killer.

    Science.gov (United States)

    Bossi, Giovanna; Trambas, Christina; Booth, Sarah; Clark, Richard; Stinchcombe, Jane; Griffiths, Gillian M

    2002-11-01

    Cytotoxic T lymphocytes (CTLs) destroy their targets by a process involving secretion of specialized granules. The interactions between CTLs and target can be very brief; nevertheless, adhesion and signaling proteins segregate into an immunological synapse. Secretion occurs in a specialized secretory domain. Use of live and fixed cell microscopy allows this secretory synapse to be visualized both temporally and spatially. The combined use of confocal and electron microscopy has produced some surprising findings, which suggest that the secretory synapse may be important both in delivering the lethal hit and in facilitating membrane transfer from target to CTL. Studies on the secretory synapse in wild-type and mutant CTLs have been used to identify proteins involved in secretion. Further clues as to the signals required for secretion are emerging from comparisons of inhibitory and activating synapses formed by natural killer cells.

  15. The Future Internet: A World of Secret Shares

    Directory of Open Access Journals (Sweden)

    William J. Buchanan

    2015-11-01

    Full Text Available The Public Key Infrastructure (PKI is crumbling, partially due to the lack of a strong understanding of how encryption actually works, but also due to weaknesses in its implementation. This paper outlines an Internet storage technique using secret sharing methods which could be used to overcome the problems inherent with PKI, while supporting new types of architectures incorporating such things as automated failover and break-glass data recovery. The paper outlines a novel architecture: SECRET, which supports a robust cloud-based infrastructure with in-built privacy and failover. In order to understand the performance overhead of SECRET, the paper outlines a range of experiments that investigate the overhead of this and other secret share methods.

  16. Bacterial Type 6 Secreted Phospholipases Play Family Feud

    OpenAIRE

    Hogan, Deborah A.; Hammond, John H.

    2013-01-01

    Secreted bacterial phospholipases play many important roles in host-pathogen interactions. In a recent study, Russell et al. (Russell et al., 2013) revealed a new role for highly conserved proteobacterial phospholipases in bacterium-bacterium interactions.

  17. TSH secreting adenoma: a rare cause of severe headache | Olt ...

    African Journals Online (AJOL)

    Considering secondary hyper thyroidism pituitary MR was performed. Pituitary MR revealed a 13x18 mm macro adenoma (Figure). The patient underwent transphenoidal surgery. Final diagnosis was TSH secreting adenoma after pathological examination. After surgery headache and hyperthyroidism were recovered

  18. Multi-group dynamic quantum secret sharing with single photons

    Energy Technology Data Exchange (ETDEWEB)

    Liu, Hongwei [School of Science and State Key Laboratory of Information Photonics and Optical Communications, Beijing University of Posts and Telecommunications, Beijing 100876 (China); Ma, Haiqiang, E-mail: hqma@bupt.edu.cn [School of Science and State Key Laboratory of Information Photonics and Optical Communications, Beijing University of Posts and Telecommunications, Beijing 100876 (China); Wei, Kejin [School of Science and State Key Laboratory of Information Photonics and Optical Communications, Beijing University of Posts and Telecommunications, Beijing 100876 (China); Yang, Xiuqing [School of Science, Beijing Jiaotong University, Beijing 100044 (China); Qu, Wenxiu; Dou, Tianqi; Chen, Yitian; Li, Ruixue; Zhu, Wu [School of Science and State Key Laboratory of Information Photonics and Optical Communications, Beijing University of Posts and Telecommunications, Beijing 100876 (China)

    2016-07-15

    In this letter, we propose a novel scheme for the realization of single-photon dynamic quantum secret sharing between a boss and three dynamic agent groups. In our system, the boss can not only choose one of these three groups to share the secret with, but also can share two sets of independent keys with two groups without redistribution. Furthermore, the security of communication is enhanced by using a control mode. Compared with previous schemes, our scheme is more flexible and will contribute to a practical application. - Highlights: • A multi-group dynamic quantum secret sharing with single photons scheme is proposed. • Any one of the groups can be chosen to share secret through controlling the polarization of photons. • Two sets of keys can be shared simultaneously without redistribution.

  19. The relationship between bone turnover and insulin sensitivity and secretion

    DEFF Research Database (Denmark)

    Frost, Morten; Balkau, Beverley; Hatunic, Mensud

    2018-01-01

    Bone metabolism appears to influence insulin secretion and sensitivity, and insulin promotes bone formation in animals, but similar evidence in humans is limited. The objectives of this study are to explore if bone turnover markers were associated with insulin secretion and sensitivity...... and to determine if bone turnover markers predict changes in insulin secretion and sensitivity. The study population encompassed 576 non-diabetic adult men with normal glucose tolerance (NGT; n=503) or impaired glucose regulation (IGR; n=73). Baseline markers of bone resorption (CTX) and formation (P1NP) were......, baseline bone turnover markers were neither associated with insulin sensitivity (assessed using hyperinsulinaemic euglycaemic clamp and OGTT) nor with insulin secretion capacity (based on IVGTT and OGTT) at baseline or at follow-up. Although inverse associations between fasting glucose and markers of bone...

  20. Detection of B-lymphocytes secreting antibodies to Dermatophagoides antigens

    DEFF Research Database (Denmark)

    Sparholt, S H; Barington, T; Schou, C

    1991-01-01

    An enzyme-linked immunospot assay (ELI-spot assay) has been established to count individual cells secreting antibodies to Dermatophagoides spp. allergens. Initial optimization of the assay was performed using Der p I-specific murine hybridoma cell lines. Inhibition with soluble purified allergen...... demonstrated that spot formation was specific. Addition of cycloheximide showed that spot formation was due to antibodies actively produced and secreted and not caused by preformed surface-associated Ig. An analogous method was used to count immunoglobulin-secreting cells (IgSC) of any specificity...... Alutard, showed that 1 week after injection, circulating allergen-specific antibody-secreting cells (AbSC) were detected with this assay. The ranges were between 1 and 13 IgM-, 2 and 20 IgG- and 20 and 55 IgA allergen-specific AbSC per 10(6) MNC. It is expected that this assay will help to understand more...

  1. Type VII secretion-mycobacteria show the way.

    NARCIS (Netherlands)

    Abdallah, A.M.; Gey van Pittius, N.C.; Champion, P.A.; Cox, J.; Luirink, S.; Vandenbroucke-Grauls, C.M.J.E.; Appelmelk, B.J.; Bitter, W.

    2007-01-01

    Recent evidence shows that mycobacteria have developed novel and specialized secretion systems for the transport of extracellular proteins across their hydrophobic, and highly impermeable, cell wall. Strikingly, mycobacterial genomes encode up to five of these transport systems. Two of these

  2. Biological activities of skin secretions of the salamander Tylototriton verrucosus.

    Science.gov (United States)

    Lai, Ren; Yang, Dong-Ming; Lee, Wen-Hui; Zhang, Yun

    2002-08-01

    Water-soluble skin secretions of salamander Tylototriton verrucosus, first described by Anderson in 1871, were studied for their biological and enzymatic activities. They were found to be toxic to mice with an intraperitoneal LD50 of 11.5 mg/kg. Using Sephadex G-75 gel filtration, it was proven that the toxic components of the secretions are proteins with molecular weights ranging from 30,000 to 50,000 Da. The secretions of T. verrucosus display a wide spectrum of antimicrobial activities and also contain both proteolytic activity and trypsin inhibitory activity. In contrast, neither hemolytic nor hemorrhagic activities were found. The secretions were determined to have phospholipase A2 activity; however, no acetylcholine esterase activity was detectable under the assay conditions.

  3. Giant machine set to probe secrets of the universe

    CERN Multimedia

    2006-01-01

    "Deep underground on the Franco-Swiss border someone will throw a switch next year to start one of the most ambitious experiments in history, probing the secrets of the universe and possibly finding new dimensions." (2/3 page)

  4. Giant machine set to probe secrets of the universe

    CERN Multimedia

    2006-01-01

    "Deep underground on the Franco-Swiss border someone will throw a switch next year to start one of the most ambitious experiments in history, probing the secrets of the universe and possibly finding new dimensions." (1 page)

  5. High-Yield Secretion of Multiple Client Proteins in Aspergillus

    Energy Technology Data Exchange (ETDEWEB)

    Segato, F.; Damasio, A. R. L.; Goncalves, T. A.; de Lucas, R. C.; Squina, F. M.; Decker, S. R.; Prade, R. A.

    2012-07-15

    Production of pure and high-yield client proteins is an important technology that addresses the need for industrial applications of enzymes as well as scientific experiments in protein chemistry and crystallization. Fungi are utilized in industrial protein production because of their ability to secrete large quantities of proteins. In this study, we engineered a high-expression-secretion vector, pEXPYR that directs proteins towards the extracellular medium in two Aspergillii host strains, examine the effect of maltose-induced over-expression and protein secretion as well as time and pH-dependent protein stability in the medium. We describe five client proteins representing a core set of hemicellulose degrading enzymes that accumulated up to 50-100 mg/L of protein. Using a recyclable genetic marker that allows serial insertion of multiple genes, simultaneous hyper-secretion of three client proteins in a single host strain was accomplished.

  6. 5 CFR 1312.27 - Top secret control.

    Science.gov (United States)

    2010-01-01

    ... classified material and will be stored in a safe under his or her control. Such material will be returned to... establish custody, record distribution, routing, receipting and destruction of Top Secret material. Top...

  7. Machine set to probe secrets of the universe

    CERN Multimedia

    Lovell, Jeremy

    2006-01-01

    "Deep underground on the Franco-Swiss border someonw will throw a switch next year to start one of the most ambitious experiments in history, probing the secrets of the universe and possibly finding new dimensions."

  8. Atrial secretion of B-type natriuretic peptide

    DEFF Research Database (Denmark)

    Goetze, Jens Peter; Friis-Hansen, Lennart; Rehfeld, Jens F

    2006-01-01

    In the normal heart, the endocrine capacity resides in the atria. Atrial myocytes express and secrete natriuretic hormones that regulate fluid homeostasis and blood pressure. But in ventricular disease, atrial natriuretic peptide (ANP) and B-type natriuretic peptide (BNP) gene expression is also...... activated in ventricular myocytes. Plasma concentrations of natriuretic peptides and their biosynthetic precursors are accordingly increased in patients with marked ventricular dysfunction. In contrast, atrial peptide secretion in ventricular disease has received less attention, and our present...

  9. Role of calcium signaling in epithelial bicarbonate secretion.

    Science.gov (United States)

    Jung, Jinsei; Lee, Min Goo

    2014-06-01

    Transepithelial bicarbonate secretion plays a key role in the maintenance of fluid and protein secretion from epithelial cells and the protection of the epithelial cell surface from various pathogens. Epithelial bicarbonate secretion is mainly under the control of cAMP and calcium signaling. While the physiological roles and molecular mechanisms of cAMP-induced bicarbonate secretion are relatively well defined, those induced by calcium signaling remain poorly understood in most epithelia. The present review summarizes the current status of knowledge on the role of calcium signaling in epithelial bicarbonate secretion. Specifically, this review introduces how cytosolic calcium signaling can increase bicarbonate secretion by regulating membrane transport proteins and how it synergizes with cAMP-induced mechanisms in epithelial cells. In addition, tissue-specific variations in the pancreas, salivary glands, intestines, bile ducts, and airways are discussed. We hope that the present report will stimulate further research into this important topic. These studies will provide the basis for future medicines for a wide spectrum of epithelial disorders including cystic fibrosis, Sjögren's syndrome, and chronic pancreatitis. Copyright © 2014 Elsevier Ltd. All rights reserved.

  10. Verifiable Rational Secret Sharing Scheme in Mobile Networks

    Directory of Open Access Journals (Sweden)

    En Zhang

    2015-01-01

    Full Text Available With the development of mobile network, lots of people now have access to mobile phones and the mobile networks give users ubiquitous connectivity. However, smart phones and tablets are poor in computational resources such as memory size, processor speed, and disk capacity. So far, all existing rational secret sharing schemes cannot be suitable for mobile networks. In this paper, we propose a verifiable rational secret sharing scheme in mobile networks. The scheme provides a noninteractively verifiable proof for the correctness of participants’ share and handshake protocol is not necessary; there is no need for certificate generation, propagation, and storage in the scheme, which is more suitable for devices with limited size and processing power; in the scheme, every participant uses her encryption on number of each round as the secret share and the dealer does not have to distribute any secret share; every participant cannot gain more by deviating the protocol, so rational participant has an incentive to abide by the protocol; finally, every participant can obtain the secret fairly (means that either everyone receives the secret, or else no one does in mobile networks. The scheme is coalition-resilient and the security of our scheme relies on a computational assumption.

  11. Xanthomonas campestris pv. vesicatoria Secretes Proteases and Xylanases via the Xps Type II Secretion System and Outer Membrane Vesicles

    Science.gov (United States)

    Solé, Magali; Scheibner, Felix; Hoffmeister, Anne-Katrin; Hartmann, Nadine; Hause, Gerd; Rother, Annekatrin; Jordan, Michael; Lautier, Martine; Arlat, Matthieu

    2015-01-01

    ABSTRACT Many plant-pathogenic bacteria utilize type II secretion (T2S) systems to secrete degradative enzymes into the extracellular milieu. T2S substrates presumably mediate the degradation of plant cell wall components during the host-pathogen interaction and thus promote bacterial virulence. Previously, the Xps-T2S system from Xanthomonas campestris pv. vesicatoria was shown to contribute to extracellular protease activity and the secretion of a virulence-associated xylanase. The identities and functions of additional T2S substrates from X. campestris pv. vesicatoria, however, are still unknown. In the present study, the analysis of 25 candidate proteins from X. campestris pv. vesicatoria led to the identification of two type II secreted predicted xylanases, a putative protease and a lipase which was previously identified as a virulence factor of X. campestris pv. vesicatoria. Studies with mutant strains revealed that the identified xylanases and the protease contribute to virulence and in planta growth of X. campestris pv. vesicatoria. When analyzed in the related pathogen X. campestris pv. campestris, several T2S substrates from X. campestris pv. vesicatoria were secreted independently of the T2S systems, presumably because of differences in the T2S substrate specificities of the two pathogens. Furthermore, in X. campestris pv. vesicatoria T2S mutants, secretion of T2S substrates was not completely absent, suggesting the contribution of additional transport systems to protein secretion. In line with this hypothesis, T2S substrates were detected in outer membrane vesicles, which were frequently observed for X. campestris pv. vesicatoria. We, therefore, propose that extracellular virulence-associated enzymes from X. campestris pv. vesicatoria are targeted to the Xps-T2S system and to outer membrane vesicles. IMPORTANCE The virulence of plant-pathogenic bacteria often depends on TS2 systems, which secrete degradative enzymes into the extracellular milieu. T2S

  12. Extracellular secretion of a recombinant therapeutic peptide by Bacillus halodurans utilizing a modified flagellin type III secretion system

    Directory of Open Access Journals (Sweden)

    Nxumalo Nolwandle P

    2011-08-01

    Full Text Available Abstract Background Through modification of the flagellin type III secretion pathway of Bacillus halodurans heterologous peptides could be secreted into the medium as flagellin fusion monomers. The stability of the secreted monomers was significantly enhanced through gene-targeted inactivation of host cell extracellular proteases. In evaluating the biotechnological potential of this extracellular secretion system an anti-viral therapeutic peptide, Enfuvirtide, was chosen. Currently, Enfuvirtide is synthesised utilizing 106 chemical steps. We used Enfuvirtide as a model system in an effort to develop a more cost-effective biological process for therapeutic peptide production. Results An attempt was made to increase the levels of the fusion peptide by two strategies, namely strain improvement through gene-targeted knock-outs, as well as vector and cassette optimization. Both approaches proved to be successful. Through chromosomal inactivation of the spo0A, lytC and lytE genes, giving rise to strain B. halodurans BhFDL05S, the secretion of recombinant peptide fusions was increased 10-fold. Cassette optimization, incorporating an expression vector pNW33N and the N- and C-terminal regions of the flagellin monomer as an in-frame peptide fusion, resulted in a further 3.5-fold increase in the secretion of recombinant peptide fusions. Conclusions The type III flagellar secretion system of B. halodurans has been shown to successfully secrete a therapeutic peptide as a heterologous flagellin fusion. Improvements to both the strain and expression cassette led to increased levels of recombinant peptide, showing promise for a biotechnological application.

  13. Expressed prostatic secretion biomarkers improve stratification of NCCN active surveillance candidates: performance of secretion capacity and TMPRSS2:ERG models.

    Science.gov (United States)

    Whelan, Christopher; Kawachi, Mark; Smith, David D; Linehan, Jennifer; Babilonia, Gail; Mejia, Rosa; Wilson, Timothy; Smith, Steven S

    2014-01-01

    Active surveillance is a viable patient option for prostate cancer provided that a clinical determination of low risk and presumably organ confined disease can be made. To standardize risk stratification schemes the NCCN (National Comprehensive Cancer Network®) provides guidelines for the active surveillance option. We determined the effectiveness of expressed prostatic secretion biomarkers for detecting occult risk factors in NCCN active surveillance candidates. Expressed prostatic secretion specimens were obtained before robot-assisted radical prostatectomy. Secretion capacity biomarkers, including total RNA and expressed prostatic secretion specimen volume, were measured by standard techniques. RNA expression biomarkers, including TXNRD1 mRNA, prostate specific antigen mRNA, TMPRSS2:ERG fusion mRNA and PCA3 mRNA, were measured by quantitative reverse-transcription polymerase chain reaction. Of the 528 patients from whom expressed prostatic secretions were collected 216 were eligible for active surveillance under NCCN guidelines. Variable selection on logistic regression identified 2 models, including one featuring types III and VI TMPRSS2:ERG variants, and one featuring 2 secretion capacity biomarkers. Of the 2 high performing models the secretion capacity model was most effective for detecting cases in this group that were up-staged or up-staged plus upgraded. It decreased the risk of up-staging in patients with a negative test almost eightfold and decreased the risk of up-staging plus upgrading about fivefold while doubling the prevalence of up-staging in the positive test group. Noninvasive expressed prostatic secretion testing may improve patient acceptance of active surveillance by dramatically reducing the presence of occult risk factors among those eligible for active surveillance under NCCN guidelines. Copyright © 2014 American Urological Association Education and Research, Inc. Published by Elsevier Inc. All rights reserved.

  14. Model for glucagon secretion by pancreatic α-cells.

    Directory of Open Access Journals (Sweden)

    Virginia González-Vélez

    Full Text Available Glucagon hormone is synthesized and released by pancreatic α-cells, one of the islet-cell types. This hormone, along with insulin, maintains blood glucose levels within the physiological range. Glucose stimulates glucagon release at low concentrations (hypoglycemia. However, the mechanisms involved in this secretion are still not completely clear. Here, using experimental calcium time series obtained in mouse pancreatic islets at low and high glucose conditions, we propose a glucagon secretion model for α-cells. Our model takes into account that the resupply of releasable granules is not only controlled by cytoplasmic Ca2+, as in other neuroendocrine and endocrine cells, but also by the level of extracellular glucose. We found that, although calcium oscillations are highly variable, the average secretion rates predicted by the model fall into the range of values reported in the literature, for both stimulated and non-stimulated conditions. For low glucose levels, the model predicts that there would be a well-controlled number of releasable granules refilled slowly from a large reserve pool, probably to ensure a secretion rate that could last for several minutes. Studying the α-cell response to the addition of insulin at low glucose, we observe that the presence of insulin reduces glucagon release by decreasing the islet Ca2+ level. This observation is in line with previous work reporting that Ca2+ dynamics, mainly frequency, is altered by insulin. Thus, the present results emphasize the main role played by Ca2+ and glucose in the control of glucagon secretion by α-cells. Our modeling approach also shows that calcium oscillations potentiate glucagon secretion as compared to constant levels of this cellular messenger. Altogether, the model sheds new light on the subcellular mechanisms involved in α-cell exocytosis, and provides a quantitative predictive tool for studying glucagon secretion modulators in physiological and pathological

  15. [Irregular secretion of prolactin in infertile women with normoprolactinemic galactorrhea].

    Science.gov (United States)

    Villanueva Díaz, Carlos A; Echavarria Sánchez, Mirna; Juárez Bengoa, Armando

    2007-02-01

    Abnormal frequency and pulse amplitud of prolactin secretion in micro and macroprolactinomas has been atributed to a dysfunctional tumoral lactotrope. Previous evidence suggests that non tumoral hyperprolactinemia is caused by a hypothalamic dysfunction. The regularity of prolactin secretion has not been studied with cuantitative methods in patients with normoprolactinemic galactorrhea (NPG) which could be considered an entity that precedes non tumoral and tumoral hyperprolactinemia. To analyze the 24-hour prolactin secretion pattern and its secretion regularity in a group of infertile women with normoprolactinemic galatorea. A transversal-comparative study was carried out in 6 infertile women with normoprolactinemic galactorrhea and 4 healthy women as controls. The 24 hour prolactin profile, the ratio night time mean concentration/daytime mean concentrattion (NM/DM ratio) and apparent entropy (Ap En, Ap En ratio) were compared in the two groups. Blunting of the nyctohemeral rythm and nocturn hyperprolactinaemia occurred in patients with normoprolactinemic galactorrhea (NPG). NM/DM ratio was lower in patients with NPG than in controls (1.28 +/- 0.25 vs. 1.75 +/- 0.05; p= 0.01). Higher irregularity of prolactin secretion was found in patients with NPG (ApEn: 0.853 +/- 0.158 vs 0.608 +/- 0.171, p=0.04; Ap En ratio: 0.839 +/- 0.11 vs 0.661 +/- 0.14; p=0.04). The irregularity of prolactin secretion in patients with NPG is not dependant on the presence of a pituitary tumour which suggests that a hypothalamic dysfunction underlies this condition. An irregular secretion and a higher daily mass production of prolactin in patients with NPG could explain both galactorrhea and infertility.

  16. Nonlinear analysis and prediction of pulsatile hormone secretion

    Energy Technology Data Exchange (ETDEWEB)

    Prank, K. [Abteilung Klinische Endokrinologie, Medizinische Hochschule Hannover, D-30623 Hannover (Germany)]|[Howard Hughes Medical Institute and Computational Neurobiology Laboratory, The Salk Institute, San Diego, California 92186-5800 (United States); Kloppstech, M. [Abteilung Klinische Endokrinologie, Medizinische Hochschule Hannover, D-30623 Hannover (Germany); Nowlan, S.J. [Howard Hughes Medical Institute and Computational Neurobiology Laboratory, The Salk Institute, San Diego, California 92186-5800 (United States); Harms, H.M.; Brabant, G.; Hesch, R. [Abteilung Klinische Endokrinologie, Medizinische Hochschule Hannover, D-30623 Hannover (Germany); Sejnowski, T.J. [Howard Hughes Medical Institute and Computational Neurobiology Laboratory, The Salk Institute, San Diego, California 92186-5800 (United States)

    1996-06-01

    Pulsatile hormone secretion is observed in almost every hormonal system. The frequency of episodic hormone release ranges from approximately 10 to 100 pulses in 24 hours. This temporal mode of secretion is an important feature of intercellular information transfer in addition to a dose-response dependent regulation. It has been demonstrated in a number of experiments that changes in the temporal pattern of pulsatile hormone secretion specifically regulate cellular and organ function and structure. Recent evidence links osteoporosis, a disease characterized by loss of bone mass and structure, to changes in the dynamics of pulsatile parathyroid hormone (PTH) secretion. In our study we applied nonlinear and linear time series prediction to characterize the secretory dynamics of PTH in both healthy human subjects and patients with osteoporosis. Osteoporotic patients appear to lack periods of high predictability found in normal humans. In contrast to patients with osteoporosis patients with hyperparathyroidism, a condition which despite sometimes reduced bone mass has a preserved bone architecture, show periods of high predictability of PTH secretion. Using stochastic surrogate data sets which match certain statistical properties of the original time series significant nonlinear determinism could be found for the PTH time series of a group of healthy subjects. Using classical nonlinear analytical techniques we could demonstrate that the irregular pattern of pulsatile PTH secretion in healthy men exhibits characteristics of deterministic chaos. Pulsatile secretion of PTH in healthy subjects seems to be a first example of nonlinear determinism in an apparently irregular hormonal rhythm in human physiology. {copyright} {ital 1996 American Institute of Physics.}

  17. Phylogenetic and functional signals in gymnosperm ovular secretions.

    Science.gov (United States)

    Nepi, Massimo; Little, Stefan; Guarnieri, Massimo; Nocentini, Daniele; Prior, Natalie; Gill, Julia; Barry Tomlinson, P; Ickert-Bond, Stefanie M; Pirone, Cary; Pacini, Ettore; von Aderkas, Patrick

    2017-11-28

    Gymnosperms are either wind-pollinated (anemophilous) or both wind- and insect-pollinated (ambophilous). Regardless of pollination mode, ovular secretions play a key role in pollen capture, germination and growth; they are likely also involved in pollinator reward. Little is known about the broad-scale diversity of ovular secretions across gymnosperms, and how these may relate to various reproductive functions. This study analyses the sugar and amino acid profiles of ovular secretions across a range of ambophilous (cycads and Gnetales) and anemophilous gymnosperms (conifers) to place them in an evolutionary context of their possible functions during reproduction. Ovular secretions from 13 species representing all five main lineages of extant gymnosperms were sampled. High-performance liquid chromatography techniques were used to measure sugar and amino acid content. Multivariate statistics were applied to assess whether there are significant differences in the chemical profiles of anemophilous and ambophilous species. Data were compared with published chemical profiles of angiosperm nectar. Chemical profiles were placed in the context of phylogenetic relationships. Total sugar concentrations were significantly higher in ovular secretions of ambophilous species than wind-pollinated taxa such as Pinaceae and Cupressophyta. Ambophilous species had lower amounts of total amino acids, and a higher proportion of non-protein amino acids compared with anemophilous lineages, and were also comparable to angiosperm nectar. Results suggest that early gymnosperms likely had ovular secretion profiles that were a mosaic of those associated with modern anemophilous and ambophilous species. Ginkgo, thought to be anemophilous, had a profile typical of ambophilous taxa, suggesting that insect pollination either exists in Gingko, but is undocumented, or that its ancestral populations were insect-pollinated. Chemical profiles of ovular secretions of ambophilous gymnosperms show a clear

  18. Human mesenchymal stem cells secrete hyaluronan-coated extracellular vesicles.

    Science.gov (United States)

    Arasu, Uma Thanigai; Kärnä, Riikka; Härkönen, Kai; Oikari, Sanna; Koistinen, Arto; Kröger, Heikki; Qu, Chengjuan; Lammi, Mikko J; Rilla, Kirsi

    2017-12-01

    Extracellular vesicles (EVs) secreted by stem cells are potential factors mediating tissue regeneration. They travel from bone marrow stem cells into damaged tissues, suggesting that they can repair tissue injuries without directly replacing parenchymal cells. We have discovered that hyaluronan (HA) synthesis is associated with the shedding of HA-coated EVs. The aim of this study was to test whether bone marrow-derived hMSCs secrete HA-coated EVs. The EVs secreted by MSCs were isolated by differential centrifugation and characterized by nanoparticle tracking analysis. Their morphology and budding mechanisms were inspected by confocal microscopy and correlative light and electron microscopy. Hyaluronan synthesis of hMSCs was induced by lipopolysaccharide and inhibited by RNA interference and 4-methylumbelliferone. It was found that the MSCs have extremely long apical and lateral HA-coated filopodia, typical for cells with an active HA secretion. Additionally, they secreted HA-coated EVs carrying mRNAs for CD44 and all HAS isoforms. The results show that stem cells have a strong intrinsic potential for HA synthesis and EV secretion, and the amount of HA carried on EVs reflects the HA content of the original cells. These results show that the secretion of HA-coated EVs by hMSCs is a general process, that may contribute to many of the mechanisms of HA-mediated tissue regeneration. Additionally, an HA coat on EVs may regulate their interactions with target cells and participate in extracellular matrix remodeling. Copyright © 2017 Elsevier B.V. All rights reserved.

  19. Secretion of Insulinotropic Proteins by Commensal Bacteria: Rewiring the Gut To Treat Diabetes▿ †

    OpenAIRE

    Duan, Faping; Curtis, Katherine L.; March, John C.

    2008-01-01

    Here, we show that commensal bacteria can stimulate intestinal epithelial cells to secrete insulin in response to glucose. Commensal strains were engineered to secrete the insulinotropic proteins GLP-1 and PDX-1. Epithelia stimulated by engineered strains and glucose secreted up to 1 ng ml−1 of insulin with no significant background secretion.

  20. Mutations in ppe38 block PE_PGRS secretion and increase virulence of Mycobacterium tuberculosis

    NARCIS (Netherlands)

    Ates, Louis S; Dippenaar, Anzaan; Ummels, Roy; Piersma, Sander R; van der Woude, Aniek D; van der Kuij, Kim; Le Chevalier, Fabien; Mata-Espinosa, Dulce; Barrios-Payán, Jorge; Marquina-Castillo, Brenda; Guapillo, Carolina; Jiménez, Connie R; Pain, Arnab; Houben, Edith N G; Warren, Robin M; Brosch, Roland; Hernández-Pando, Rogelio; Bitter, Wilbert

    Mycobacterium tuberculosis requires a large number of secreted and exported proteins for its virulence, immune modulation and nutrient uptake. Most of these proteins are transported by the different type VII secretion systems1,2. The most recently evolved type VII secretion system, ESX-5, secretes

  1. Influence of Flavonoids on Mechanism of Modulation of Insulin Secretion.

    Science.gov (United States)

    Soares, Juliana Mikaelly Dias; Pereira Leal, Ana Ediléia Barbosa; Silva, Juliane Cabral; Almeida, Jackson R G S; de Oliveira, Helinando Pequeno

    2017-01-01

    The development of alternatives for insulin secretion control in vivo or in vitro represents an important aspect to be investigated. In this direction, natural products have been progressively explored with this aim. In particular, flavonoids are potential candidates to act as insulin secretagogue. To study the influence of flavonoid on overall modulation mechanisms of insulin secretion. The research was conducted in the following databases and platforms: PubMed, Scopus, ISI Web of Knowledge, SciELO, LILACS, and ScienceDirect, and the MeSH terms used for the search were flavonoids, flavones, islets of Langerhans, and insulin-secreting cells. Twelve articles were included and represent the basis of discussion on mechanisms of insulin secretion of flavonoids. Papers in ISI Web of Knowledge were in number of 1, Scopus 44, PubMed 264, ScienceDirect 511, and no papers from LILACS and SciELO databases. According to the literature, the majority of flavonoid subclasses can modulate insulin secretion through several pathways, in an indication that corresponding molecule is a potential candidate for active materials to be applied in the treatment of diabetes. The action of natural products on insulin secretion represents an important investigation topic due to their importance in the diabetes controlIn addition to their typical antioxidant properties, flavonoids contribute to the insulin secretionThe modulation of insulin secretion is induced by flavonoids according to different mechanisms. Abbreviations used: KATP channels: ATP-sensitive K+ channels, GLUT4: Glucose transporter 4, ERK1/2: Extracellular signal-regulated protein kinases 1 and 2, L-VDCCs: L-type voltage-dependent Ca+2 channels, GLUT1: Glucose transporter 1, AMPK: Adenosine monophosphate-activated protein kinase, PTP1B: Protein tyrosine phosphatase 1B, GLUT2: Glucose transporter 2, cAMP: Cyclic adenosine monophosphate, PKA: Protein kinase A, PTK: Protein tyrosine kinase, CaMK II: Ca2+/calmodulin

  2. Traffic Noise Exposure Increases Gastric Pepsin Secretion in Rat

    Directory of Open Access Journals (Sweden)

    Azam Moslehi

    2016-04-01

    Full Text Available Noise is considered as one of the most severe sources of environmental and workplace constraints. Many noise effects are well known on immune function, hormonal levels, cardiovascular and respiratory systems. In this study, our aim is to evaluate the effects of traffic noise exposure on basal and stimulated gastric pepsin secretion. 48 male rats were exposed to traffic noise (86 dB for a short term of (8h/ day for 1 day and a long term of (8h/ day for 7, 14, 21 and 28 days as well as a control group. The gastric contents were collected by the wash-out technique. Pepsin secretion was measured by employing the Anson method. Histological studies were carried out on the epithelial layer. The corticosteroid hormone was measured in the serum for the stress augmentation. The present finding indicated no changes in pepsin secretion content in the short term, but in the 14 and 21 days traffic noise exposure, basal gastric pepsin secretion increased markedly compared to the control group. Histological results showed that the number of oxyntic glands and cell nuclei decreased in comparison with the control group while the thickness of the epithelial layer increases. In addition, the corticosterone levels increase in all groups in comparison with the control. It seems that the increase of gastric pepsin secretion is due to the description and translation processes in the peptic cells and needs enough time for completion.

  3. Androecia in two Clusia species: development, structure and resin secretion.

    Science.gov (United States)

    Sá-Haiad, B; Silva, C P; Paula, R C V; Rocha, J F; Machado, S R

    2015-07-01

    Clusia fluminensis and C. lanceolata are dioecious shrubs having resiniferous flowers with strongly distinct androecia. The aim of this study was to investigate the development and anatomy of their androecia and the ultrastructure, histochemistry and secretory process of their androecium resin glands, examining whether the cellular aspects of resin secretion differed between these two morphologically distinct androecia. Stamens differ, being free in C. fluminensis and clustered in a synandrium in C. lanceolata. Staminode sterility is due to the undifferentiated nature of the anthers in C. lanceolata and degeneration of meiocytes and anther indehiscence in C. fluminensis. Resin is produced in subepidermal cavities and canals with wide lumens. In the secretory stage, epithelial cells present sinuous walls, voluminous nuclei, polymorphic plastids associated with periplastidial reticulum, mitochondria, oil bodies, multivesicular bodies, endoplasmic reticulum and dictyosomes. The resin is released through rupture points on the distal surface of stamens and staminodes, associated with disrupted cavities and canals. Our results show morphological diversity associated with functional similarity. Also, a secretion pattern shared by the two species includes initiation of the secretory process in young floral buds, compartmentalisation of the secretion in pre-anthesis buds and release of secretions at anthesis. Cellular aspects of resin secretion in these species are quite similar, as are the chemical identities of the main components of the floral resins of the genus. © 2015 German Botanical Society and The Royal Botanical Society of the Netherlands.

  4. Traffic Noise Exposure Increases Gastric Pepsin Secretion in Rat.

    Science.gov (United States)

    Moslehi, Azam; Nabavizadeh, Fatemeh; Keshavarz, Mansoor; Rouhbakhsh, Nematollah; Sotudeh, Masoud; Salimi, Ehsan; Barzegar Behrooz, Amir

    2016-03-01

    Noise is considered as one of the most severe sources of environmental and workplace constraints. Many noise effects are well known on immune function, hormonal levels, cardiovascular and respiratory systems. In this study, our aim is to evaluate the effects of traffic noise exposure on basal and stimulated gastric pepsin secretion. 48 male rats were exposed to traffic noise (86 dB) for a short term of (8h/day for 1 day) and a long term of (8h/day for 7, 14, 21 and 28 days) as well as a control group. The gastric contents were collected by the wash-out technique. Pepsin secretion was measured by employing the Anson method. Histological studies were carried out on the epithelial layer. The corticosteroid hormone was measured in the serum for the stress augmentation. The present finding indicated no changes in pepsin secretion content in the short term, but in the 14 and 21 days traffic noise exposure, basal gastric pepsin secretion increased markedly compared to the control group. Histological results showed that the number of oxyntic glands and cell nuclei decreased in comparison with the control group while the thickness of the epithelial layer increases. In addition, the corticosterone levels increase in all groups in comparison with the control. It seems that the increase of gastric pepsin secretion is due to the description and translation processes in the peptic cells and needs enough time for completion.

  5. Zinc-secreting Paneth cells studied by ZP fluorescence.

    Science.gov (United States)

    Giblin, Leonard J; Chang, Christopher J; Bentley, Anthony F; Frederickson, Cathleen; Lippard, Stephen J; Frederickson, Christopher J

    2006-03-01

    We have used a new family of zinc-specific-responsive fluorescent dyes (ZPs) to study the sequestration and secretion of zinc from Paneth cells, which are located in the bases of the crypts of Lieberkühn within the rat small intestine. Vivid ZP fluorescence zinc staining of Paneth cell secretory granules is seen in both cryostat sections and isolated crypts, providing firm evidence for a pool of labile (rapidly exchangeable) zinc within these cells. We further demonstrate that this ionic zinc pool is secreted under physiological conditions. In vivo stimulation of the small intestine by IP injection of the secretagogue pilocarpine results in discrete zinc staining within the lumens of subsequently isolated crypts, concomitant with a decrease in the zinc staining of Paneth cell granules located within the same crypts. In contrast, the secretion of zinc into the lumens of isolated crypts stimulated in vitro with either carbachol or LPS (lipopolysaccharide) is not observed. However, a distinct change in Paneth cell morphology, suggesting attempted secretion, is seen in response to the direct application of cholinergics but not LPS. These findings suggest that zinc is coreleased with other Paneth cell anti-microbials, and that the intact intestine is necessary for secretion into the crypt lumen.

  6. Insulin secretion abnormalities in exocrine pancreatic sufficient cystic fibrosis patients.

    Science.gov (United States)

    Wooldridge, Jamie L; Szczesniak, Rhonda D; Fenchel, Matthew C; Elder, Deborah A

    2015-11-01

    The aim of this study is to assess insulin secretion in pediatric cystic fibrosis (CF) patients with exocrine pancreatic sufficiency. Glucose and insulin responses during an oral glucose tolerance test (OGTT) were measured in 146 CF patients. Patients were divided into exocrine sufficient (CF-PS) and insufficient (CF-PI) groups based on pancreatic enzyme usage and fecal elastase. A reference group included healthy, non-diabetic subjects. All CF groups showed reduced insulin secretion as measured by insulinogenic index. The CF-PS patients had normal glucose tolerance. There was a direct correlation between BMI z-score and insulin area under the curve. Patients with CF have reduced insulin secretion during an OGTT regardless of exocrine pancreatic status. The abnormal insulin secretion in all CF patients may predispose them for glucose intolerance, particularly when challenged by inflammation, infection, or nutritional deficiency. In addition, the diminished insulin secretion may contribute to increased catabolism. Lastly, the CF-related diabetes (CFRD) screening guidelines should be followed by all CF patients regardless of pancreatic status. Copyright © 2015 European Cystic Fibrosis Society. Published by Elsevier B.V. All rights reserved.

  7. Peroxicretion: a novel secretion pathway in the eukaryotic cell

    Directory of Open Access Journals (Sweden)

    Luesken Francisca A

    2009-05-01

    Full Text Available Abstract Background Enzyme production in microbial cells has been limited to secreted enzymes or intracellular enzymes followed by expensive down stream processing. Extracellular enzymes consists mainly of hydrolases while intracellular enzymes exhibit a much broader diversity. If these intracellular enzymes could be secreted by the cell the potential of industrial applications of enzymes would be enlarged. Therefore a novel secretion pathway for intracellular proteins was developed, using peroxisomes as secretion vesicles. Results Peroxisomes were decorated with a Golgi derived v-SNARE using a peroxisomal membrane protein as an anchor. This allowed the peroxisomes to fuse with the plasma membrane. Intracellular proteins were transported into the peroxisomes by adding a peroxisomal import signal (SKL tag. The proteins which were imported in the peroxisomes, were released into the extra-cellular space through this artificial secretion pathway which was designated peroxicretion. This concept was supported by electron microscopy studies. Conclusion Our results demonstrate that it is possible to reroute the intracellular trafficking of vesicles by changing the localisation of SNARE molecules, this approach can be used in in vivo biological studies to clarify the different control mechanisms regulating intracellular membrane trafficking. In addition we demonstrate peroxicretion of a diverse set of intracellular proteins. Therefore, we anticipate that the concept of peroxicretion may revolutionize the production of intracellular proteins from fungi and other microbial cells, as well as from mammalian cells.

  8. Maturation processes and structures of small secreted peptides in plants

    Directory of Open Access Journals (Sweden)

    Ryo eTabata

    2014-07-01

    Full Text Available In the past decade, small secreted peptides have proven to be essential for various aspects of plant growth and development, including the maintenance of certain stem cell populations. Most small secreted peptides identified in plants to date are recognised by membrane-localized receptor kinases, the largest family of receptor proteins in the plant genome. This peptide-receptor interaction is essential for initiating intracellular signalling cascades. Small secreted peptides often undergo post-translational modifications and proteolytic processing to generate the mature peptides. Recent studies suggest that, in contrast to the situation in mammals, the proteolytic processing of plant peptides involves a number of complex steps. Furthermore, NMR-based structural analysis demonstrated that post-translational modifications induce the conformational changes needed for full activity. In this mini review, we summarise recent advances in our understanding of how small secreted peptides are modified and processed into biologically active peptides and describe the mature structures of small secreted peptides in plants.

  9. A subcellular model of glucose-stimulated pancreatic insulin secretion.

    Science.gov (United States)

    Pedersen, Morten Gram; Corradin, Alberto; Toffolo, Gianna M; Cobelli, Claudio

    2008-10-13

    When glucose is raised from a basal to stimulating level, the pancreatic islets respond with a typical biphasic insulin secretion pattern. Moreover, the pancreas is able to recognize the rate of change of the glucose concentration. We present a relatively simple model of insulin secretion from pancreatic beta-cells, yet founded on solid physiological grounds and capable of reproducing a series of secretion patterns from perfused pancreases as well as from stimulated islets. The model includes the notion of distinct pools of granules as well as mechanisms such as mobilization, priming, exocytosis and kiss-and-run. Based on experimental data, we suggest that the individual beta-cells activate at different glucose concentrations. The model reproduces most of the data it was tested against very well, and can therefore serve as a general model of glucose-stimulated insulin secretion. Simulations predict that the effect of an increased frequency of kiss-and-run exocytotic events is a reduction in insulin secretion without modification of the qualitative pattern. Our model also appears to be the first physiology-based one to reproduce the staircase experiment, which underlies 'derivative control', i.e. the pancreatic capacity of measuring the rate of change of the glucose concentration.

  10. Suppression of Insulin Production and Secretion by a Decretin Hormone

    Science.gov (United States)

    Alfa, Ronald W.; Park, Sangbin; Skelly, Kathleen-Rose; Poffenberger, Gregory; Jain, Nimit; Gu, Xueying; Kockel, Lutz; Wang, Jing; Liu, Yinghua; Powers, Alvin C.; Kim, Seung K.

    2015-01-01

    SUMMARY Decretins, hormones induced by fasting that suppress insulin production and secretion, have been postulated from classical human metabolic studies. From genetic screens, we identified Drosophila Limostatin (Lst), a peptide hormone that suppresses insulin secretion. Lst is induced by nutrient restriction in gut-associated endocrine cells. limostatin deficiency led to hyperinsulinemia, hypoglycemia and excess adiposity. A conserved 15-residue polypeptide encoded by limostatin suppressed secretion by insulin-producing cells. Targeted knockdown of CG9918, a Drosophila orthologue of Neuromedin U receptors (NMUR), in insulin-producing cells phenocopied limostatin deficiency, and attenuated insulin suppression by purified Lst, suggesting CG9918 encodes an Lst receptor. NMUR1 is expressed in islet β-cells, and purified NMU suppresses insulin secretion from human islets. A human mutant NMU variant that co-segregates with familial early-onset obesity and hyperinsulinemia fails to suppress insulin secretion. We propose Lst as an index member of an ancient hormone class called decretins, which suppress insulin output. PMID:25651184

  11. Bottleneck in secretion of α-amylase in Bacillus subtilis.

    Science.gov (United States)

    Yan, Shaomin; Wu, Guang

    2017-07-19

    Amylase plays an important role in biotechnology industries, and Gram-positive bacterium Bacillus subtilis is a major host to produce heterogeneous α-amylases. However, the secretion stress limits the high yield of α-amylase in B. subtilis although huge efforts have been made to address this secretion bottleneck. In this question-oriented review, every effort is made to answer the following questions, which look simple but are long-standing, through reviewing of literature: (1) Does α-amylase need a specific and dedicated chaperone? (2) What signal sequence does CsaA recognize? (3) Does CsaA require ATP for its operation? (4) Does an unfolded α-amylase is less soluble than a folded one? (5) Does α-amylase aggregate before transporting through Sec secretion system? (6) Is α-amylase sufficient stable to prevent itself from misfolding? (7) Does α-amylase need more disulfide bonds to be stabilized? (8) Which secretion system does PrsA pass through? (9) Is PrsA ATP-dependent? (10) Is PrsA reused after folding of α-amylase? (11) What is the fate of PrsA? (12) Is trigger factor (TF) ATP-dependent? The literature review suggests that not only the most of those questions are still open to answers but also it is necessary to calculate ATP budget in order to better understand how B. subtilis uses its energy for production and secretion.

  12. Ultrastructure and oil secretion in Hiptage sericea Hook

    Directory of Open Access Journals (Sweden)

    K. Arumugasamy

    2014-01-01

    Full Text Available The oil secreting glands of Hiptage sericea Hook. consist of three regions: epithelial, sub-epithelial and sub-glandular. In early stages, the oil secreting cells are characterized by the presence of plastids with starch grains and electron translucent vesicles, mitochondria, rER, polysomes, small vacuoles, numerous lipid bodies and well-defined nucleus with nucleolus. Later, the accumulation of plastoglobuli and inclusion bodies occur in the matrix of the plastid. Tubular, smooth endoplasmic reticulum begins to appear in the cytoplasm. With the onset of secretion, the osmiophilic contents of plastids which appear as electron dense, round droplets move-into cytoplasm and often occur in the region of the plasmalemma invaginations. However, in matured glands the lipid bodies disappear from the cytoplasm. The size of the vacuoles increases and are filled with electron opaque substance. Similar substances are also found in the sub-cuticular spaces as well as outside the cuticle.

  13. Hyperosmolar solutions stimulate mucus secretion in the ferret trachea.

    Science.gov (United States)

    Kishioka, Chikako; Okamoto, Kosuke; Kim, Jung-Soo; Rubin, Bruce K

    2003-07-01

    Inhalation of hypertonic saline or mannitol solutions acutely increases mucociliary clearance. Because increased clearance is often coupled with increased mucus secretion, we hypothesized that hyperosmolar agents would stimulate mucus secretion. The isolated tracheae of healthy young adult ferrets were studied in a basic research laboratory. We demonstrated that there was a dose-dependent increase in mucin secretion by enzyme-linked lectin assay after incubation with 1.69 g/dL (597 mOsm/L), 3.69 g/dL (1,192 mOsm/L), 5.69 g/dL (1,823 mOsm/L), and 10.69 g/dL (3,612 mOsm/L) of saline solution over Krebs-Henseleit solution control (288 mOsm/L) [p care when used for patients with mucus hypersecretion and impaired mucus clearance.

  14. Noncoherent capacity of secret-key agreement with public discussion

    KAUST Repository

    Agrawal, Anurag

    2011-09-01

    We study the noncoherent capacity of secret-key agreement with public discussion over independent identically distributed (i.i.d.) Rayleigh fading wireless channels, where neither the sender nor the receivers have access to instantaneous channel state information (CSI). We present two results. At high signal-to-noise ratio (SNR), the secret-key capacity is bounded in SNR, regardless of the number of antennas at each terminal. Second, for a system with a single antenna at both the legitimate and the eavesdropper terminals and an arbitrary number of transmit antennas, the secret-key capacity-achieving input distribution is discrete, with a finite number of mass points. Numerically we observe that at low SNR, the capacity achieving distribution has two mass points with one of them at the origin. © 2011 IEEE.

  15. Security of Linear Secret-Sharing Schemes Against Mass Surveillance

    DEFF Research Database (Denmark)

    Giacomelli, Irene; Olimid, Ruxandra; Ranellucci, Samuel

    2015-01-01

    Following the line of work presented recently by Bellare, Paterson and Rogaway, we formalize and investigate the resistance of linear secret-sharing schemes to mass surveillance. This primitive is widely used to design IT systems in the modern computer world, and often it is implemented by a prop......Following the line of work presented recently by Bellare, Paterson and Rogaway, we formalize and investigate the resistance of linear secret-sharing schemes to mass surveillance. This primitive is widely used to design IT systems in the modern computer world, and often it is implemented...... by a proprietary code that the provider (“big brother”) could manipulate to covertly violate the privacy of the users (by implementing Algorithm-Substitution Attacks or ASAs). First, we formalize the security notion that expresses the goal of big brother and prove that for any linear secret-sharing scheme...

  16. Secret Sharing over Fast-Fading MIMO Wiretap Channels

    Directory of Open Access Journals (Sweden)

    Bloch Matthieu

    2009-01-01

    Full Text Available Secret sharing over the fast-fading MIMO wiretap channel is considered. A source and a destination try to share secret information over a fast-fading MIMO channel in the presence of an eavesdropper who also makes channel observations that are different from but correlated to those made by the destination. An interactive, authenticated public channel with unlimited capacity is available to the source and destination for the secret sharing process. This situation is a special case of the "channel model with wiretapper" considered by Ahlswede and Csiszár. An extension of their result to continuous channel alphabets is employed to evaluate the key capacity of the fast-fading MIMO wiretap channel. The effects of spatial dimensionality provided by the use of multiple antennas at the source, destination, and eavesdropper are then investigated.

  17. Targeting development of incretin-producing cells increases insulin secretion

    DEFF Research Database (Denmark)

    Petersen, Natalia; Reimann, Frank; van Es, Johan H

    2015-01-01

    Glucagon-like peptide-1-based (GLP-1-based) therapies improve glycemic control in patients with type 2 diabetes. While these agents augment insulin secretion, they do not mimic the physiological meal-related rise and fall of GLP-1 concentrations. Here, we tested the hypothesis that increasing...... systems and augmented glucose-stimulated GLP-1 secretion. In a high-fat diet-fed mouse model of impaired glucose tolerance and type 2 diabetes, dibenzazepine administration increased L cell numbers in the intestine, improved the early insulin response to glucose, and restored glucose tolerance....... Dibenzazepine also increased K cell numbers, resulting in increased gastric inhibitory polypeptide (GIP) secretion. Using a GLP-1 receptor antagonist, we determined that the insulinotropic effect of dibenzazepine was mediated through an increase in GLP-1 signaling. Together, our data indicate that modulation...

  18. Regulation of renin secretion by renal juxtaglomerular cells

    DEFF Research Database (Denmark)

    Friis, Ulla G; Madsen, Kirsten; Stubbe, Jane

    2013-01-01

    , neurotransmitters, and cell swelling converge on the stimulatory cyclic AMP (cAMP) pathway. Renin secretion is attenuated in mice deficient in beta-adrenoceptors, prostaglandin E(2)-EP4 receptors, Gsα protein, and adenylyl cyclases 5 and 6. Phosphodiesterases (PDE) 3 and 4 degrade cAMP in JG cells, and PDE3...... is inhibited by cyclic GMP (cGMP) and couples the cGMP pathway to the cAMP pathway. Cyclic AMP enhances K(+)-current in JG cells and is permissive for secretion by stabilizing membrane potential far from threshold that activates L-type voltage-gated calcium channels. Intracellular calcium paradoxically...... inhibits renin secretion likely through attenuated formation and enhanced degradation of cAMP; by activation of chloride currents and interaction with calcineurin. Connexin 40 is necessary for localization of JG cells in the vascular wall and for pressure- and macula densa-dependent suppression of renin...

  19. Incretins, insulin secretion and Type 2 diabetes mellitus

    DEFF Research Database (Denmark)

    Vilsbøll, T; Holst, Jens Juul

    2004-01-01

    the genes encoding their receptors have been deleted. In patients with Type 2 diabetes, the incretin effect is either greatly impaired or absent, and it is assumed that this could contribute to the inability of these patients to adjust their insulin secretion to their needs. In studies of the mechanism...... of the impaired incretin effect in Type 2 diabetic patients, it has been found that the secretion of GIP is generally normal, whereas the secretion of GLP-1 is reduced, presumably as a consequence of the diabetic state. It might be of even greater importance that the effect of GLP-1 is preserved whereas...... the effect of GIP is severely impaired. The impaired GIP effect seems to have a genetic background, but could be aggravated by the diabetic state. The preserved effect of GLP-1 has inspired attempts to treat Type 2 diabetes with GLP-1 or analogues thereof, and intravenous GLP-1 administration has been shown...

  20. Recurrent menorrhagia in an adolescent with a platelet secretion defect.

    Science.gov (United States)

    Santos, Xiomara M; Bercaw-Pratt, Jennifer L; Yee, Donald L; Dietrich, Jennifer E

    2011-04-01

    Although von Willebrand disease is the most common inherited bleeding disorder, platelet function disorders are less well recognized as a cause of bleeding. We report a case of menorrhagia caused by an unsuspected platelet secretion defect. A 13-year-old Asian female, with unknown family history, presented with menorrhagia not responsive to intravenous conjugated estrogens, requiring transfusion of 7 units of packed red blood cells. Initial screening tests for bleeding disorders were normal; however, due to high clinical suspicion, further specific testing with platelet aggregometry was performed, which revealed a platelet secretion defect. The prevalence of platelet secretion defects in adolescents with menorrhagia is unknown, but may be higher than currently recognized. When screening tests are normal, yet suspicion remains high for an underlying hemostatic disorder, platelet aggregometry must be performed. Copyright © 2011 North American Society for Pediatric and Adolescent Gynecology. Published by Elsevier Inc. All rights reserved.

  1. Possible modulatory effect of endogenous islet catecholamines on insulin secretion

    Directory of Open Access Journals (Sweden)

    Gagliardino Juan J

    2001-10-01

    Full Text Available Abstract Background The possible participation of endogenous islet catecholamines (CAs in the control of insulin secretion was tested. Methods Glucose-induced insulin secretion was measured in the presence of 3-Iodo-L-Tyrosine (MIT, a specific inhibitor of tyrosine-hydroxylase activity, in fresh and precultured islets isolated from normal rats. Incubated islets were also used to measure CAs release in the presence of low and high glucose, and the effect of α2-(yohimbine [Y] and idazoxan [I] and α1-adrenergic antagonists (prazosin [P] and terazosin [T] upon insulin secretion elicited by high glucose. Results Fresh islets incubated with 16.7 mM glucose released significantly more insulin in the presence of 1 μM MIT (6.66 ± 0.39 vs 5.01 ± 0.43 ng/islet/h, p Conclusion Our results suggest that islet-originated CAs directly modulate insulin release in a paracrine manner.

  2. Secret Sharing over Fast-Fading MIMO Wiretap Channels

    Directory of Open Access Journals (Sweden)

    Tan F. Wong

    2009-01-01

    Full Text Available Secret sharing over the fast-fading MIMO wiretap channel is considered. A source and a destination try to share secret information over a fast-fading MIMO channel in the presence of an eavesdropper who also makes channel observations that are different from but correlated to those made by the destination. An interactive, authenticated public channel with unlimited capacity is available to the source and destination for the secret sharing process. This situation is a special case of the “channel model with wiretapper” considered by Ahlswede and Csiszár. An extension of their result to continuous channel alphabets is employed to evaluate the key capacity of the fast-fading MIMO wiretap channel. The effects of spatial dimensionality provided by the use of multiple antennas at the source, destination, and eavesdropper are then investigated.

  3. Ionizing radiation induces tumor cell lysyl oxidase secretion

    DEFF Research Database (Denmark)

    Shen, Colette J; Sharma, Ashish; Vuong, Dinh-Van

    2014-01-01

    metalloproteinases, among others, to promote tumor progression. Lysyl oxidase is known to play an important role in hypoxia-dependent cancer cell dissemination and metastasis. Here, we investigated the effects of IR on the expression and secretion of lysyl oxidase (LOX) from tumor cells. METHODS: LOX-secretion along...... with enzymatic activity was investigated in multiple tumor cell lines in response to irradiation. Transwell migration assays were performed to evaluate invasive capacity of naive tumor cells in response to IR-induced LOX. In vivo studies for confirming IR-enhanced LOX were performed employing...... immunohistochemistry of tumor tissues and ex vivo analysis of murine blood serum derived from locally irradiated A549-derived tumor xenografts. RESULTS: LOX was secreted in a dose dependent way from several tumor cell lines in response to irradiation. IR did not increase LOX-transcription but induced LOX...

  4. Prelysosomal Compartments in the Unconventional Secretion of Amyloidogenic Seeds

    DEFF Research Database (Denmark)

    Borland, Helena; Vilhardt, Frederik

    2017-01-01

    . The ESCRT-mediated import of cytosolic amyloid into late endosomal exosomes, a known vehicle of transmission of macromolecules between cells, is also reviewed. Finally, mechanisms of lysosomal dysfunction, deficiency, and exocytosis are exemplified in the context of genetically identified risk factors......A mechanistic link between neuron-to-neuron transmission of secreted amyloid and propagation of protein malconformation cytopathology and disease has recently been uncovered in animal models. An enormous interest in the unconventional secretion of amyloids from neurons has followed. Amphisomes......, mainly for Parkinson's disease. Exocytosis of prelysosomal or lysosomal organelles is a last resort for clearance of cytotoxic material and alleviates cytopathy. However, they also represent a vehicle for the concentration, posttranslational modification, and secretion of amyloid seeds....

  5. Endogenous ACh tonically stimulates ANP secretion in rat atria.

    Science.gov (United States)

    Kim, Hye Yoom; Cho, Kyung Woo; Xu, Dong Yuan; Kang, Dae Gill; Lee, Ho Sub

    2013-10-01

    Exogenous acetylcholine (ACh) is known to stimulate atrial natriuretic peptide (ANP) secretion concomitantly with a decrease in atrial pulse pressure. However, the role of intrinsic ACh in the regulation of ANP secretion remains unknown. Recently, it was shown that nonneuronal and neuronal ACh is present in the cardiac atria. From this finding we hypothesize that endogenously released ACh is involved in the regulation of ANP secretion in an autocrine or paracrine manner in the atria. Experiments were performed in isolated beating rat atria. ANP was measured using radioimmunoassay. To increase the availability of the ACh in the extracellular space of the atrium, its degradation was inhibited with an inhibitor of acetylcholinesterase. Acetylcholinesterase inhibition with physostigmine increased ANP secretion concomitantly with a decrease in atrial dynamics in a concentration-dependent manner. Inhibitors of M2 muscarinic ACh receptor (mAChR), methoctramine, and ACh-activated K(+) (KACh(+)) channels, tertiapin-Q, abolished the physostigmine-induced changes. The effects were not observed in the atria from rats treated with pertussis toxin. Furthermore, the physostigmine-induced effects were attenuated by an inhibitor of high-affinity choline transporter, hemicholinium-3, which is a rate-limiting step of ACh synthesis. Inhibitors of the mAChR signaling pathway and ACh synthesis also attenuated the basal levels of ANP secretion and accentuated atrial dynamics. These findings suggest that endogenously released ACh tonically stimulates ANP secretion from atrial cardiomyocytes via activation of M2 mAChR-Gi/o-KACh(+) channel signaling. It is also suggested that the ACh-ANP signaling is implicated in cardiac physiology and pathophysiology.

  6. FluG affects secretion in colonies of Aspergillus niger.

    Science.gov (United States)

    Wang, Fengfeng; Krijgsheld, Pauline; Hulsman, Marc; de Bekker, Charissa; Müller, Wally H; Reinders, Marcel; de Vries, Ronald P; Wösten, Han A B

    2015-01-01

    Colonies of Aspergillus niger are characterized by zonal heterogeneity in growth, sporulation, gene expression and secretion. For instance, the glucoamylase gene glaA is more highly expressed at the periphery of colonies when compared to the center. As a consequence, its encoded protein GlaA is mainly secreted at the outer part of the colony. Here, multiple copies of amyR were introduced in A. niger. Most transformants over-expressing this regulatory gene of amylolytic genes still displayed heterogeneous glaA expression and GlaA secretion. However, heterogeneity was abolished in transformant UU-A001.13 by expressing glaA and secreting GlaA throughout the mycelium. Sequencing the genome of UU-A001.13 revealed that transformation had been accompanied by deletion of part of the fluG gene and disrupting its 3' end by integration of a transformation vector. Inactivation of fluG in the wild-type background of A. niger also resulted in breakdown of starch under the whole colony. Asexual development of the ∆fluG strain was not affected, unlike what was previously shown in Aspergillus nidulans. Genes encoding proteins with a signal sequence for secretion, including part of the amylolytic genes, were more often downregulated in the central zone of maltose-grown ∆fluG colonies and upregulated in the intermediate part and periphery when compared to the wild-type. Together, these data indicate that FluG of A. niger is a repressor of secretion.

  7. Increased amyloidogenic secretion in cerebellar granule cells undergoing apoptosis

    Science.gov (United States)

    Galli, Cinzia; Piccini, Alessandra; Ciotti, Maria Teresa; Castellani, Loriana; Calissano, Pietro; Zaccheo, Damiano; Tabaton, Massimo

    1998-01-01

    Some clues suggest that neuronal damage induces a secondary change of amyloid β protein (Aβ) metabolism. We investigated this possibility by analyzing the secretion of Aβ and processing of its precursor protein (amyloid precursor protein, APP) in an in vitro model of neuronal apoptosis. Primary cultures of rat cerebellar granule neurons were metabolically labeled with [35S]methionine. Apoptosis was induced by shifting extracellular KCl concentration from 25 mM to 5 mM for 6 h. Control and apoptotic neurons were then subjected to depolarization-stimulated secretion. Constitutive and stimulated secretion media and cell lysates were immunoprecipitated with antibodies recognizing regions of Aβ, full-length APP, α- and β-APP secreted forms. Immunoprecipitated proteins were separated by SDS/PAGE and quantitated with a PhosphorImager densitometer. Although intracellular full-length APP was not significantly changed after apoptosis, the monomeric and oligomeric forms of 4-kDa Aβ were 3-fold higher in depolarization-stimulated secretion compared with control neurons. Such increments were paralleled by a corresponding increase of the β-APPs/α-APPs ratio in apoptotic secretion. Immunofluorescence studies performed with an antibody recognizing an epitope located in the Aβ sequence showed that the Aβ signal observed in the cytoplasm and in the Golgi apparatus of control neurons is uniformly redistributed in the condensed cytoplasm of apoptotic cells. These studies indicate that neuronal apoptosis is associated with a significant increase of metabolic products derived from β-secretase cleavage and suggest that an overproduction of Aβ may be the consequence of neuronal damage from various causes. PMID:9448317

  8. Kinetics of alpha-amylase secretion in Aspergillus oryzae

    DEFF Research Database (Denmark)

    Henriksen, Anne Laurence Santerre; Carlsen, Morten; Bang de, H.

    1999-01-01

    Pulse and pulse-chase experiments have been performed to study L-[S-35] methionine incorporation and protein secretion kinetics in Aspergillus oryzae. Pulse experiments confirmed the mechanism of methionine uptake reported previously for Penicillium chrysogenum (Benko et al., 1967). Pulse......-chase experiments were carried out to investigate the alpha-amylase secretion kinetics in A. oryzae. No unglycosylated alpha-amylase was detected neither intracellularly nor extracellularly demonstrating that glycosylation was not the rate controlling step in the secretory pathway. The pulse chase experiments...

  9. ["Secret causes": causality and determinism in the classical age].

    Science.gov (United States)

    Cléro, Jean-Pierre

    2014-01-01

    The notion of the "secret cause", which appears in many classical texts is tied to a particular practice of science and a conception of its methods where the "law" finds itself at the center of the nexus. If certain phenomena appear to escape the law, one is obliged to amend the law through the introduction of a series of "small equations." If the calculation of probabilities is deployed, this is to precisely reveal causes which are, at their origin, secret, but which will gradually become less so and eventually conform to laws.

  10. Functional characterization of serotonin receptor subtypes in human duodenal secretion

    DEFF Research Database (Denmark)

    Engelmann, Bodil Elisabeth; Bindslev, Niels; Poulsen, Steen Seier

    2006-01-01

    Serotonin (5-HT) stimulates ion secretion in the gastrointestinal tract and the sensitivity for 5-HT might be altered in dyspeptic patients infected with Helicobacter pylori. The purpose of the present study was to characterize the 5-HT-induced electrogenic ion transport in the duodenum of dyspep......Serotonin (5-HT) stimulates ion secretion in the gastrointestinal tract and the sensitivity for 5-HT might be altered in dyspeptic patients infected with Helicobacter pylori. The purpose of the present study was to characterize the 5-HT-induced electrogenic ion transport in the duodenum...

  11. Characterization of the macula densa stimulus for renin secretion

    DEFF Research Database (Denmark)

    Lorenz, J N; Weihprecht, H; Schnermann, J

    1990-01-01

    These studies utilize the isolated perfused rabbit juxtaglomerular apparatus (JGA) to study the macula densa signal for renin secretion in the absence of the confounding influences of intravascular pressure and renal nerve activity. In the first experimental series, JGAs were perfused alternately...... with high- and low-NaCl solutions to determine the reversibility of the renin response to changes in NaCl concentration. Compared with high-NaCl controls, perfusion with a low-NaCl solution resulted in a fivefold increase in renin secretion rate (RSR) [2.1-10.0 nano-Goldblatt hog units (n...

  12. Laparoscopic Partial Adrenalectomy for Bilateral Cortisol-secreting Adenomas

    Directory of Open Access Journals (Sweden)

    Jeffrey P. Domino

    2007-04-01

    Full Text Available Bilateral cortisol-secreting adenomas are a rare cause of Cushing's syndrome. We report a case of a 35-year-old woman who presented with ACTH-independent Cushing's syndrome and bilateral adrenal adenomas. Adrenal venous sampling confirmed both adenomas to be hyper-secreting cortisol. She underwent bilateral laparoscopic adrenalectomy; total right and partial left adrenalectomies. At 2-year follow-up, she is maintained on low-dose fludrocortisone and hydrocortisone, and without recurrence of hypercorticolism. Laparoscopic partial adrenalectomy is a feasible option for this rare condition; however, long-term follow-up is needed to determine her total independence from steroid usage.

  13. Effects of antimycin A and 2-deoxyglucose on secretion in human platelets. Differential inhibition of the secretion of acid hydrolases and adenine nucleotides.

    Science.gov (United States)

    Holmsen, H; Robkin, L; Day, H J

    1979-08-15

    1. Shape change, aggregation and secretion of dense-granule constituents in platelets differ in their dependence on cellular energy metabolism. The possibility that such a difference also exists between secretion of dense-granule constituents and acid hydrolases was investigated. 2. Human platelets were incubated with [(14)C]adenine in plasma, and then washed and resuspended in salt solutions. The effects of incubating the cells with antimycin A and 2-deoxyglucose on the concentrations of [(14)C]ATP, ADP, AMP, IMP and inosine plus hypoxanthine and on thrombin-induced secretion of ATP plus ADP and acid hydrolases were studied. The metabolic inhibitors only affected (14)C-labelled nucleotides, whereas thrombin only liberated unlabelled ATP and ADP. 3. The extent of secretion decreased progressively with time during incubation with the metabolic inhibitors. At any time the secretion of acid hydrolases, beta-N-acetylglucosaminidase, beta-glucuronidase and beta-galactosidase was inhibited to a greater extent than secretion of ATP plus ADP (dense-granule secretion). 4. Incubation with the metabolic inhibitors shifted the log (dose)-response relationship to higher thrombin concentrations, and with a greater shift for acid hydrolase secretion than for dense-granule secretion. 5. Antimycin, when present alone, caused a marked decrease in the rate of acid hydrolase secretion, but had no effect on dense-granule secretion. 6. These results further support the view that acid hydrolase secretion and dense-granule secretion are separate processes with different requirements for ATP energy. Acid hydrolase secretion, but not dense-granule secretion, appears to depend on a simultaneous rapid generation of ATP, which can be accomplished by oxidative, but not by glycolytic, ATP production.

  14. DPP4 gene variation affects GLP-1 secretion, insulin secretion, and glucose tolerance in humans with high body adiposity

    DEFF Research Database (Denmark)

    Böhm, Anja; Wagner, Robert; Machicao, Fausto

    2017-01-01

    , inter-individual variance in the responsiveness to DPP-4 inhibitors was reported. Thus, we asked whether genetic variation in the DPP4 gene affects incretin levels, insulin secretion, and glucose tolerance in participants of the TÜbingen Family study for type-2 diabetes (TÜF). RESEARCH DESIGN......-BMI interaction effects were detected (p = 0.8). After stratification for body fat content, the SNP negatively affected glucose-stimulated GLP-1 levels (p = 0.0229), insulin secretion (p = 0.0061), and glucose tolerance (p = 0.0208) in subjects with high body fat content only. CONCLUSIONS: A common variant, i.......e., SNP rs6741949, in the DPP4 gene interacts with body adiposity and negatively affects glucose-stimulated GLP-1 levels, insulin secretion, and glucose tolerance. Whether this SNP underlies the reported inter-individual variance in responsiveness to DPP-4 inhibitors, at least in subjects with high body...

  15. Quantitative secretion and maximal secretion capacity of retinol, b-carotene and a-tocopherol into cows' milk

    DEFF Research Database (Denmark)

    Jensen, Søren Krogh; Kjær, Anna Kirstin Bjørnbak; Hermansen, John Erik

    1999-01-01

    Changes in the contents of retinol, a-tocopherol and b-carotene in plasma, milk and milk fat from 38 Holstein-Friesian cows were followed during their ®rst lactation, and the quantitative and kinetic relationships for secretion of a-tocopherol and b-carotene from blood into milk were determined......, retinol and b-carotene concentration also differed according to sire. The concentrations of all three vitamins in milk and milk fat differed according to sire and stage of lactation. Furthermore, the total secretion of retinol, a-tocopherol and b-carotene into milk (expressed as mg}d) differed with sire...... and stage of lactation. The quantitative secretion of a-tocopherol and b-carotene from blood into milk followed Michaelis±Menten kinetics for active transport across membranes. Values of maximum secretory capacity Vmax and the half-rate constant Km for both a-tocopherol and b-carotene varied according...

  16. On the diversity-multiplexing tradeoff of secret-key agreement over multiple-antenna channels

    KAUST Repository

    Zorgui, Marwen

    2014-09-01

    We consider secret-key agreement with public discussion over Rayleigh fading quasi-static channels. First, the secret-key diversity gain and the secret-key multiplexing gain are defined. Then, the secret-key diversity multiplexing tradeoff (DMT) is established. The eavesdropper is shown to \\'steal\\' only transmit antennas. We show that likewise the DMT without secrecy constraint, the secret-key DMT is the same either with or without full channel state information (CSI) at the transmitter (CSI-T). This insensitivity of secret-key DMT toward CSI-T highlights a fundamental difference between secret-key agreement and the wiretap channel whose secret DMT depends crucially on CSI-T. Several secret-key DMT-achieving schemes are presented in case of full CSI-T.

  17. Fatty Acid Composition of Different Collections of Coprinoid Mushrooms (Agaricomycetes) and Their Nutritional and Medicinal Values.

    Science.gov (United States)

    Badalyan, Susanna M

    2016-01-01

    Qualitative and quantitative analyses of the free fatty acid (FA) composition of ethanol extracts obtained from mycelia of 30 genetically identified strains of 18 species of coprinoid mushrooms (CMs) or coprini belonging to clades Coprinellus (C. bisporus, C. curtus, C. disseminatus, C. domesticus, C. ellisii, C.flocculosus, C. micaceus, C. aff radians I, C. aff radians II, C. aff radians III, C. xanthothrix, Coprinellus sp. 4), Coprinopsis (C. cinerea, C. cothurnata, C. gonophylla, C. lagopides, C. maysoidispora, C. strossmayeri), and Coprinus (C. comatus) were evaluated using gas chromatography. Two unsaturated (lineolic, oleic) and 3 saturated (palmitic, stearic, myristic) FAs were detected in the tested mycelial samples. Amounts of unsaturated FAs prevailed over saturated FAs. Lineolic and palmitic acids were detected in all the species in high quantities (up to 64.73% and 53.82%, respectively). All 5 FAs were found in the Coprinus clade (family Agaricaceae) as opposed to the Coprinellus and Coprinopsis clades (family Psathyrellaceae). Based on the unsaturated and saturated FA content, the species from the clade Coprinellus were different from Coprinopsis species and C. comatus. The obtained data confirm that palmitic, linoleic, and stearic acids are predominant in mushrooms, including coprini. The presence of large amounts of unsaturated FAs in the tested coprini make them an option for further studies to formulate novel mushroom-based dietary supplements and nutriceuticals.

  18. Serotonergic involvement in stress-induced vasopressin and oxytocin secretion

    DEFF Research Database (Denmark)

    Jørgensen, Henrik; Knigge, Ulrich; Kjaer, Andreas

    2002-01-01

    OBJECTIVE: To investigate the involvement of serotonin (5-hydroxytryptamine - 5-HT) receptors in mediation of stress-induced arginine vasopressin (AVP) and oxytocin (OT) secretion in male rats. DESIGN: Experiments on laboratory rats with control groups. METHODS: Different stress paradigms were...

  19. Le CERN va tenter de percer les secrets

    CERN Multimedia

    2005-01-01

    CERN will try to drill the secrets of neutrinos by setting up an experiment on a large scale. This one should begin at CERN in May 2006: neutrinos are considered by the physicians as the most imperceptible particles in the Universe

  20. Degrading capability and activity of extracellular xylanase secreted ...

    African Journals Online (AJOL)

    The natural lignocellulose degrading capabilities of extracellular enzyme secreted by a composite microbial system XDC-2 were studied. Peptone cellulose solution (PCS) medium was beneficial to the degradation of lignocellulosic materials and ATCC 1053 medium promoted enzyme production of XDC-2. The exocellular ...