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Sample records for rokiikoon nick oliveri

  1. Nick Laws | NREL

    Science.gov (United States)

    -3238 Nick's primary work at NREL revolves around REopt - a techno-economic optimization model for , Nick is continuously adapting the REopt model to accommodate the latest technologies, market factors spiral and the impact of utility rate structures on the adoption of residential solar photovoltaics and

  2. antispasmodic actions of the leaves of daniellia oliveri

    African Journals Online (AJOL)

    The leaves of the plant Daniellia oliveri are usually prescribed traditionally in northern Nigeria for varieties of gastrointestinal complaints. The antispasmodic activity of the n-butanol soluble part of the aqueous portion of the ethanolic extract was evaluated in vitro on isolated guinea pig ileum against three spasmogen; ...

  3. Antihyperglycaemic effect of aqueous extract of Daniella oliveri and ...

    African Journals Online (AJOL)

    JTEkanem

    results indicate that the decoction of the roots of S. latifolius and D. oliveri has .... Protein Determination. Protein content of the liver tissues was determined by the Biuret method as described by Gornal et al.14 using Bovine Serum. Albumin ...

  4. Daniellia oliveri As A Fodder Tree For Small Ruminant And The ...

    African Journals Online (AJOL)

    Daniellia oliveri was examined as a potential fodder for small ruminant, using nine castrated and ruminally fistulated West African Dwarf sheep (29 kg BW) to determine rumen ammonia and nutrient digestibility. Dried leaves of Daniella oliveri were offered at two levels (25% and 50% of DMI) as supplement to a basal hay ...

  5. Studying repair of a single protein-bound nick in vivo using the Flp-nick system

    DEFF Research Database (Denmark)

    Nielsen, Ida; Andersen, Anni Hangaard; Bjergbæk, Lotte

    2012-01-01

    The Flp-nick system is a simple in vivo system developed for studying the cellular responses to a protein-bound nick at a single genomic site in the budding yeast Saccharomyces cerevisiae. The Flp-nick system takes advantage of a mutant Flp recombinase that can introduce a nick at a specific Flp ...

  6. Roman for unge - og deres voksne - fra Nick Hornby

    DEFF Research Database (Denmark)

    Haarder, Jon Helt

    2007-01-01

    Nick Hornby: SLAM. Oversat fra engelsk. af Jan Hansen. 280 sider, 249 kr. Lindhardt og Ringhof. Fire stjerner Udgivelsesdato: 071207......Nick Hornby: SLAM. Oversat fra engelsk. af Jan Hansen. 280 sider, 249 kr. Lindhardt og Ringhof. Fire stjerner Udgivelsesdato: 071207...

  7. Isolation and evaluation of antiglycation potential of polyalthic acid (furano-terpene from Daniella oliveri

    Directory of Open Access Journals (Sweden)

    Olubunmi Atolani

    2014-12-01

    Full Text Available A furano-diterpene (polyalthic acid was isolated as a major stable compound for the first time from the oleoresin of the Daniella oliveri of the family Caesalpiniacea through column chromatography fractionation. Polyalthic acid was characterized using data obtained from EIMS, HREIMS, ESI-MS, MALDI-MS as well as 1D and 2D NMR and it was evaluated for its potential to inhibit the formation of advanced glycation end-products (AGEs using a standard in vitro antiglycation procedure. Polyalthic acid indicated a negative antiglycation potential compared to standard inhibitor that has 85% inhibition, which is an indication that polyalthic acid may not contribute to the antiglycation activity of the plant as acclaimed in folkloric medicine. The negative antiglycation observed could indicate that the polyalthic acid could trigger glycation, thereby subjecting users to various degrees of complications. The bioactivity evaluation on molinspiration evaluator indicated that polyalthic acid could be a potential drug candidate. The biological and chemical insights gained on polyalthic acid provide a good basis for future research. Keywords: Daniella oliveria, Polyalthic acid, Furano-diterpene, Antiglycation, Matrix assisted laser desorption/ionization (MALDI

  8. Distinct genetic control of homologous recombination repair of Cas9-induced double-strand breaks, nicks and paired nicks

    NARCIS (Netherlands)

    Vriend, Lianne E. M.; Prakash, Rohit; Chen, Chun-Chin; Vanoli, Fabio; Cavallo, Francesca; Zhang, Yu; Jasin, Maria; Krawczyk, Przemek M.

    2016-01-01

    DNA double-strand breaks (DSBs) are known to be powerful inducers of homologous recombination (HR), but single-strand breaks (nicks) have also been shown to trigger HR. Both DSB- and nick-induced HR ((nick)HR) are exploited in advanced genome-engineering approaches based on the bacterial RNA-guided

  9. Otiorhynchus oliveri N.SP.DI CURCULIONIDE DELLA SPAGNA E NOTE SUL GRUPPO DI O.andarensis REITTER,1913(Coleoptera)

    Institute of Scientific and Technical Information of China (English)

    MASSIMO MEREGALLI

    1990-01-01

    <正> Otiorhynchus oliveri,n.sp.of Curculionid from Spain and notes on some taxa of the O.andarensis Reitter,1913,group.The new species belongs to the group of O.andarensis Reitter and ischaracterized by the deeper and denser punctures,the male metatibia lacking spinose processes and genitalia.Type locality:Catòute Mount,in Leon province,on rocky limestone outcrops at 1,900 metres a.s.l.Someremarks on the chorology and bioecology of other species of the group are also provided.

  10. Population genetic structure of the endemic rosewoods Dalbergia cochinchinensis and D. oliveri at a regional scale reflects the Indochinese landscape and life-history traits

    DEFF Research Database (Denmark)

    Hartvig, Ida; So, Thea; Changtragoon, Suchitra

    2018-01-01

    of the distribution area, particularly in Cambodia. We suggest that this pattern is ancient, reflecting the demographic history of the species and possible location of refugia during earlier time periods with limited forest cover, which was supported by signs of old genetic bottlenecks. The D. oliveri populations had......Indochina is a biodiversity hot spot and harbors a high number of endemic species, most of which are poorly studied. This study explores the genetic structure and reproductive system of the threatened endemic timber species Dalbergia cochinchinensis and Dalbergia oliveri using microsatellite data...... from populations across Indochina and relates it to landscape characteristics and life-history traits. We found that the major water bodies in the region, Mekong and Tonle Sap, represented barriers to gene flow and that higher levels of genetic diversity were found in populations in the center...

  11. Essays on the future in honor of Nick Metropolis

    CERN Document Server

    Rota, Gian-Carlo

    2000-01-01

    This collection represents a unique undertaking in scientific publishing to honor Nick Metropolis. Nick was the last survivor of the World War II Manhattan Project in Los Alamos, and was an important member of the Los Alamos national Laboratory until his death in October, 1999. In this volume, some of the leading scientists and humanists of our time have contributed essays related to their respective disciplines, exploring various aspects of future developments in science and society, philosophy, national security, nuclear power, pure and applied mathematics, physics and biology, particle physics, computing, and information science.

  12. Firewood yield and profitability of a traditional Daniellia oliveri short-rotation coppice on fallow lands in Benin

    International Nuclear Information System (INIS)

    Avohou, T. Hermane; Houehounha, Remy; Glele-Kakai, Romain; Assogbadjo, Achille Ephrem; Sinsin, Brice

    2011-01-01

    Sub-Saharan Africa has a great diversity of local coppicing species which are exploited in traditional short coppice systems for firewood. Biomass yield and profitability of these systems as well as their responses to silvicultural improvement are little known. This study evaluated the firewood yield and the profitability of a traditional Daniellia oliveri short-rotation coppice on fallow lands in central Benin. Two weed management options were considered: (1) the weedy option, usually practiced by locals, which experienced grass competition and bushfires, and (2) the weed-free option, which consisted in periodic removal of grasses and other species. Destructive measurements and allometric equations were used to estimate biomass yield in 12 plots over 42 months. A cost-benefit analysis model based on the net present value and the benefit-cost ratio was used to compare the profitability of the two management options. Biomass accumulation rate averaged 1.08 ± 0.20 tonnes of dry matter ha -1 year -1 (t DM ha -1 year -1 ) in weedy conditions. Weed removal improved 3.5 times this rate in weed-free plots (3.83 ± 0.47 t DM ha -1 year -1 ). After 42 months, total biomass reached 3.67 ± 0.65 t DM ha -1 in weedy plots and 11.63 ± 0.76 t DM ha -1 in weed-free plots. Most of the biomass (≥88%) was marketable in local markets. Coppice exploitation was profitable after 24 months for both management options. Weed removal improved the profits three times. A sensitivity analysis showed that both options were still profitable with up to 25% increase of labour and transport costs, 25% decrease of biomass price and 12% increase of the discount rate. (author)

  13. Nick Park pani Wallace'ile ja Gromitile libaküüliku needuse / Mart Rummo

    Index Scriptorium Estoniae

    Rummo, Mart

    2005-01-01

    Uus animafilm "Wallace ja Gromit : Libaküüliku needus" ("Wallace & Gromit : The Curse of the Were-Rabbit") : režissöör Nick Park : Suurbritannia 2005. Selle loojast ja tema loometeest. Lisatud : Nick Park (filmograafia, auhindu), "Teised Nick Parkist" (Lauri Kaare, Mait Laas, Riho Unt)

  14. DNA fragments assembly based on nicking enzyme system.

    Directory of Open Access Journals (Sweden)

    Rui-Yan Wang

    Full Text Available A couple of DNA ligation-independent cloning (LIC methods have been reported to meet various requirements in metabolic engineering and synthetic biology. The principle of LIC is the assembly of multiple overlapping DNA fragments by single-stranded (ss DNA overlaps annealing. Here we present a method to generate single-stranded DNA overlaps based on Nicking Endonucleases (NEases for LIC, the method was termed NE-LIC. Factors related to cloning efficiency were optimized in this study. This NE-LIC allows generating 3'-end or 5'-end ss DNA overlaps of various lengths for fragments assembly. We demonstrated that the 10 bp/15 bp overlaps had the highest DNA fragments assembling efficiency, while 5 bp/10 bp overlaps showed the highest efficiency when T4 DNA ligase was added. Its advantage over Sequence and Ligation Independent Cloning (SLIC and Uracil-Specific Excision Reagent (USER was obvious. The mechanism can be applied to many other LIC strategies. Finally, the NEases based LIC (NE-LIC was successfully applied to assemble a pathway of six gene fragments responsible for synthesizing microbial poly-3-hydroxybutyrate (PHB.

  15. Bypass of a Nick by the Replisome of Bacteriophage T7*

    Science.gov (United States)

    Zhu, Bin; Lee, Seung-Joo; Richardson, Charles C.

    2011-01-01

    DNA polymerase and DNA helicase are essential components of DNA replication. The helicase unwinds duplex DNA to provide single-stranded templates for DNA synthesis by the DNA polymerase. In bacteriophage T7, movement of either the DNA helicase or the DNA polymerase alone terminates upon encountering a nick in duplex DNA. Using a minicircular DNA, we show that the helicase·polymerase complex can bypass a nick, albeit at reduced efficiency of 7%, on the non-template strand to continue rolling circle DNA synthesis. A gap in the non-template strand cannot be bypassed. The efficiency of bypass synthesis depends on the DNA sequence downstream of the nick. A nick on the template strand cannot be bypassed. Addition of T7 single-stranded DNA-binding protein to the complex stimulates nick bypass 2-fold. We propose that the association of helicase with the polymerase prevents dissociation of the helicase upon encountering a nick, allowing the helicase to continue unwinding of the duplex downstream of the nick. PMID:21701044

  16. Bypass of a nick by the replisome of bacteriophage T7.

    Science.gov (United States)

    Zhu, Bin; Lee, Seung-Joo; Richardson, Charles C

    2011-08-12

    DNA polymerase and DNA helicase are essential components of DNA replication. The helicase unwinds duplex DNA to provide single-stranded templates for DNA synthesis by the DNA polymerase. In bacteriophage T7, movement of either the DNA helicase or the DNA polymerase alone terminates upon encountering a nick in duplex DNA. Using a minicircular DNA, we show that the helicase · polymerase complex can bypass a nick, albeit at reduced efficiency of 7%, on the non-template strand to continue rolling circle DNA synthesis. A gap in the non-template strand cannot be bypassed. The efficiency of bypass synthesis depends on the DNA sequence downstream of the nick. A nick on the template strand cannot be bypassed. Addition of T7 single-stranded DNA-binding protein to the complex stimulates nick bypass 2-fold. We propose that the association of helicase with the polymerase prevents dissociation of the helicase upon encountering a nick, allowing the helicase to continue unwinding of the duplex downstream of the nick.

  17. NCYM promotes calpain-mediated Myc-nick production in human MYCN-amplified neuroblastoma cells

    Energy Technology Data Exchange (ETDEWEB)

    Shoji, Wataru [Division of Biochemistry and Innovative Cancer Therapeutics and Children' s Cancer Research Center, Chiba Cancer Center Research Institute, 666-2 Nitona, Chuo-ku, Chiba 260-8717 (Japan); Department of Pediatric Surgery, Graduate School of Medicine, Tohoku University, Sendai 980-8574 (Japan); Suenaga, Yusuke, E-mail: ysuenaga@chiba-cc.jp [Division of Biochemistry and Innovative Cancer Therapeutics and Children' s Cancer Research Center, Chiba Cancer Center Research Institute, 666-2 Nitona, Chuo-ku, Chiba 260-8717 (Japan); Cancer Genome Center, Chiba Cancer Center Research Institute, 666-2 Nitona, Chuo-ku, Chiba 260-8717 (Japan); Kaneko, Yoshiki; Islam, S.M. Rafiqul; Alagu, Jennifer [Division of Biochemistry and Innovative Cancer Therapeutics and Children' s Cancer Research Center, Chiba Cancer Center Research Institute, 666-2 Nitona, Chuo-ku, Chiba 260-8717 (Japan); Yokoi, Sana [Cancer Genome Center, Chiba Cancer Center Research Institute, 666-2 Nitona, Chuo-ku, Chiba 260-8717 (Japan); Nio, Masaki [Department of Pediatric Surgery, Graduate School of Medicine, Tohoku University, Sendai 980-8574 (Japan); Nakagawara, Akira, E-mail: nakagawara-a@koseikan.jp [Division of Biochemistry and Innovative Cancer Therapeutics and Children' s Cancer Research Center, Chiba Cancer Center Research Institute, 666-2 Nitona, Chuo-ku, Chiba 260-8717 (Japan)

    2015-06-05

    NCYM is a cis-antisense gene of MYCN and is amplified in human neuroblastomas. High NCYM expression is associated with poor prognoses, and the NCYM protein stabilizes MYCN to promote proliferation of neuroblastoma cells. However, the molecular mechanisms of NCYM in the regulation of cell survival have remained poorly characterized. Here we show that NCYM promotes cleavage of MYCN to produce the anti-apoptotic protein, Myc-nick, both in vitro and in vivo. NCYM and Myc-nick were induced at G2/M phase, and NCYM knockdown induced apoptotic cell death accompanied by Myc-nick downregulation. These results reveal a novel function of NCYM as a regulator of Myc-nick production in human neuroblastomas. - Highlights: • NCYM promotes cleavages of MYC and MYCN to produce Myc-nick in vitro. • NCYM increases Myc-nick production in MYCN-amplified neuroblastoma cells. • NCYM knockdown decreases Myc-nick production and induces apoptosis at G2/M phase.

  18. NCYM promotes calpain-mediated Myc-nick production in human MYCN-amplified neuroblastoma cells

    International Nuclear Information System (INIS)

    Shoji, Wataru; Suenaga, Yusuke; Kaneko, Yoshiki; Islam, S.M. Rafiqul; Alagu, Jennifer; Yokoi, Sana; Nio, Masaki; Nakagawara, Akira

    2015-01-01

    NCYM is a cis-antisense gene of MYCN and is amplified in human neuroblastomas. High NCYM expression is associated with poor prognoses, and the NCYM protein stabilizes MYCN to promote proliferation of neuroblastoma cells. However, the molecular mechanisms of NCYM in the regulation of cell survival have remained poorly characterized. Here we show that NCYM promotes cleavage of MYCN to produce the anti-apoptotic protein, Myc-nick, both in vitro and in vivo. NCYM and Myc-nick were induced at G2/M phase, and NCYM knockdown induced apoptotic cell death accompanied by Myc-nick downregulation. These results reveal a novel function of NCYM as a regulator of Myc-nick production in human neuroblastomas. - Highlights: • NCYM promotes cleavages of MYC and MYCN to produce Myc-nick in vitro. • NCYM increases Myc-nick production in MYCN-amplified neuroblastoma cells. • NCYM knockdown decreases Myc-nick production and induces apoptosis at G2/M phase

  19. Nick Ransford: 'There is a job to do so let's get on with it'.

    Science.gov (United States)

    Ransford, Nick; Doherty, Ruth

    2014-03-01

    Nick Ransford is a consultant in special care dentistry at Birmingham Community Healthcare NHS Trust. He has over 25 years' experience working with adults with disabilities and medical conditions across the spectrum.

  20. Pangahävitaja Leeson : "Ma kahetsen" / Nick Leeson ; interv. Argo Ideon

    Index Scriptorium Estoniae

    Leeson, Nick

    2007-01-01

    Briti börsimaakler Nick Leeson, kes pankrotistas Baringsi panga ning hävitas 17 miljardit krooni, selgitab, miks ta ei suutnud oma tegevust lõpetada ka siis kui kahjud olid kasvanud juba hiigelsuureks

  1. Footprinting of Chlorella virus DNA ligase bound at a nick in duplex DNA.

    Science.gov (United States)

    Odell, M; Shuman, S

    1999-05-14

    The 298-amino acid ATP-dependent DNA ligase of Chlorella virus PBCV-1 is the smallest eukaryotic DNA ligase known. The enzyme has intrinsic specificity for binding to nicked duplex DNA. To delineate the ligase-DNA interface, we have footprinted the enzyme binding site on DNA and the DNA binding site on ligase. The size of the exonuclease III footprint of ligase bound a single nick in duplex DNA is 19-21 nucleotides. The footprint is asymmetric, extending 8-9 nucleotides on the 3'-OH side of the nick and 11-12 nucleotides on the 5'-phosphate side. The 5'-phosphate moiety is essential for the binding of Chlorella virus ligase to nicked DNA. Here we show that the 3'-OH moiety is not required for nick recognition. The Chlorella virus ligase binds to a nicked ligand containing 2',3'-dideoxy and 5'-phosphate termini, but cannot catalyze adenylation of the 5'-end. Hence, the 3'-OH is important for step 2 chemistry even though it is not itself chemically transformed during DNA-adenylate formation. A 2'-OH cannot substitute for the essential 3'-OH in adenylation at a nick or even in strand closure at a preadenylated nick. The protein side of the ligase-DNA interface was probed by limited proteolysis of ligase with trypsin and chymotrypsin in the presence and absence of nicked DNA. Protease accessible sites are clustered within a short segment from amino acids 210-225 located distal to conserved motif V. The ligase is protected from proteolysis by nicked DNA. Protease cleavage of the native enzyme prior to DNA addition results in loss of DNA binding. These results suggest a bipartite domain structure in which the interdomain segment either comprises part of the DNA binding site or undergoes a conformational change upon DNA binding. The domain structure of Chlorella virus ligase inferred from the solution experiments is consistent with the structure of T7 DNA ligase determined by x-ray crystallography.

  2. Perspectives on the ‘lens of risk’ interview series: interview with Nick Pidgeon

    NARCIS (Netherlands)

    Heyman, B.; Brown, P.

    2012-01-01

    This article is the first in a series which will appear in 2012 in the special issue series Health Care Through the `Lens of Risk'. It provides a quasi-verbatim transcript of an interview with Nick Pidgeon, one of the main contributors to the social science component of The Royal Society Risk Report

  3. An ultrasensitive colorimeter assay strategy for p53 mutation assisted by nicking endonuclease signal amplification.

    Science.gov (United States)

    Lin, Zhenyu; Yang, Weiqiang; Zhang, Guiyun; Liu, Qida; Qiu, Bin; Cai, Zongwei; Chen, Guonan

    2011-08-28

    A novel catalytic colorimetric assay assisted by nicking endonuclease signal amplification (NESA) was developed. With the signal amplification, the detection limit of the p53 target gene can be as low as 1 pM, which is nearly 5 orders of magnitude lower than that of other previously reported colorimetric DNA detection strategies based on catalytic DNAzyme.

  4. Securing wilderness landscapes in South Africa : Nick Steele, private wildlife conservancies and saving rhinos

    NARCIS (Netherlands)

    Wels,; H.,

    2015-01-01

    Private wildlife conservation is booming business in South Africa! Nick Steele stood at the cradle of this development in the politically turbulent 1970s and 1980s, by stimulating farmers in Natal (now KwaZulu-Natal) to pool resources in order to restore wilderness landscapes, but at the same time

  5. Siim Nestor soovitab : Jus Fine ja DJ Hype. Mutant ja Nick Luscombe. Kuurorti kogumik / Siim Nestor

    Index Scriptorium Estoniae

    Nestor, Siim, 1974-

    2005-01-01

    Inglise drum'n'bass-staar DJ Hype 17. märtsil klubis Privé üritusel "Jus' Fine". Inglise diskor Nick Luscombe üritusel "Mutant" 18. märtsil klubis Privé. Plaadifirma Kuurortrecords esitleb 18. märtsil oma uut kogumikplaati Pirita jõe suudmes asuval laeval Monica

  6. Divalent metal ion differentially regulates the sequential nicking reactions of the GIY-YIG homing endonuclease I-BmoI.

    Directory of Open Access Journals (Sweden)

    Benjamin P Kleinstiver

    Full Text Available Homing endonucleases are site-specific DNA endonucleases that function as mobile genetic elements by introducing double-strand breaks or nicks at defined locations. Of the major families of homing endonucleases, the modular GIY-YIG endonucleases are least understood in terms of mechanism. The GIY-YIG homing endonuclease I-BmoI generates a double-strand break by sequential nicking reactions during which the single active site of the GIY-YIG nuclease domain must undergo a substantial reorganization. Here, we show that divalent metal ion plays a significant role in regulating the two independent nicking reactions by I-BmoI. Rate constant determination for each nicking reaction revealed that limiting divalent metal ion has a greater impact on the second strand than the first strand nicking reaction. We also show that substrate mutations within the I-BmoI cleavage site can modulate the first strand nicking reaction over a 314-fold range. Additionally, in-gel DNA footprinting with mutant substrates and modeling of an I-BmoI-substrate complex suggest that amino acid contacts to a critical GC-2 base pair are required to induce a bottom-strand distortion that likely directs conformational changes for reaction progress. Collectively, our data implies mechanistic roles for divalent metal ion and substrate bases, suggesting that divalent metal ion facilitates the re-positioning of the GIY-YIG nuclease domain between sequential nicking reactions.

  7. Nick Hornby’s A Long Way Down (2005 in the Transition from Book to Movie

    Directory of Open Access Journals (Sweden)

    Ömercan Tüm

    2015-12-01

    Full Text Available Nick Hornby’s A Long Way Down (2005 in the Transition from Book to movie Abstract Challenging and profound, Nick Hornby’s novel A Long Way Down (2005 is the story of four people failing to commit suicide. The protagonists are caught in an intricate web of relationships, disappointments and missed chances on their one-way journey to understand that “The cure for unhappiness is happiness” (Elizabeth McCracken. This paper aims at demonstrating that 2014 movie version directed by Pascal Chaumeil fails to capture the essence of the book and resorts to a number of radical changes which are only supposed to attract a larger audience, but do not necessarily send the same message as the novel.

  8. Nick translation detection in situ of cellular DNA strand break induced by radiation

    International Nuclear Information System (INIS)

    Maehara, Y.; Anai, H.; Kusumoto, T.; Sakaguchi, Y.; Sugimachi, K.

    1989-01-01

    DNA strand break in HeLa cells induced by radiation was detected using the in situ nick translation method. The cells were exposed to radiation of 3, 6, 12, 18, and 24 Gy in Lab-Tek tissue culture chamber/slides and were fixed with ethanol/acetic acid on the slide glass. The break sites in DNA were translated artificially in the presence of Escherichia coli DNA polymerase I and [ 3 H]-labeled dTTP. Autoradiographic observation was made of the level of break sites in the DNA. The DNA strand break appeared even with a 3 Gy exposure, increased 8.6 times at 24 Gy compared with the control cells, and this level correlated reciprocally to change in cell viability. This nick translation method provides a rapid in situ assay for determining radiation-induced DNA damage of cultured cells, in a semi-quantitative manner

  9. Trapping and breaking of in vivo nicked DNA during pulsed-field gel electrophoresis

    Science.gov (United States)

    Khan, Sharik R.; Kuzminov, Andrei

    2013-01-01

    Pulsed field gel electrophoresis (PFGE) offers a high-resolution approach to quantify chromosomal fragmentation in bacteria, measured as percent of chromosomal DNA entering the gel. The degree of separation in PFG depends upon the size of DNA, as well as various conditions of electrophoresis, such as electric field strength (FS), time of electrophoresis, switch time and buffer composition. Here we describe a new parameter, the structural integrity of the sample DNA itself, that influences its migration through PFGs. We show that sub-chromosomal fragments containing both spontaneous and DNA damage-induced nicks are prone to breakage during PFGE. Such breakage at single strand interruptions results in artefactual decrease in molecular weight of linear DNA making accurate determination of the number of double strand breaks difficult. While breakage of nicked sub-chromosomal fragments is FS-independent, some high molecular weight sub-chromosomal fragments are also trapped within wells under the standard PFGE conditions. This trapping can be minimized by lowering the field strength and increasing the time of electrophoresis. We discuss how breakage of nicked DNA may be mechanistically linked to trapping. Our results suggest how to optimize conditions for PFGE when quantifying chromosomal fragmentation induced by DNA damage. PMID:23770235

  10. Myc-nick: a cytoplasmic cleavage product of Myc that promotes alpha-tubulin acetylation and cell differentiation.

    Science.gov (United States)

    Conacci-Sorrell, Maralice; Ngouenet, Celine; Eisenman, Robert N

    2010-08-06

    The Myc oncoprotein family comprises transcription factors that control multiple cellular functions and are widely involved in oncogenesis. Here we report the identification of Myc-nick, a cytoplasmic form of Myc generated by calpain-dependent proteolysis at lysine 298 of full-length Myc. Myc-nick retains conserved Myc box regions but lacks nuclear localization signals and the bHLHZ domain essential for heterodimerization with Max and DNA binding. Myc-nick induces alpha-tubulin acetylation and altered cell morphology by recruiting histone acetyltransferase GCN5 to microtubules. During muscle differentiation, while the levels of full-length Myc diminish, Myc-nick and acetylated alpha-tubulin levels are increased. Ectopic expression of Myc-nick accelerates myoblast fusion, triggers the expression of myogenic markers, and permits Myc-deficient fibroblasts to transdifferentiate in response to MyoD. We propose that the cleavage of Myc by calpain abrogates the transcriptional inhibition of differentiation by full-length Myc and generates Myc-nick, a driver of cytoplasmic reorganization and differentiation. Copyright 2010 Elsevier Inc. All rights reserved.

  11. The singer and the song: Nick Cave and the archetypal function of the cover version

    OpenAIRE

    Wiseman-Trowse, Nathan J B

    2013-01-01

    Throughout his career, from The Boys Next Door, through The Birthday Party, and with The Bad Seeds, Australian singer / songwriter Nick Cave has balanced his own set of creative voices alongside those of others through his choice of cover versions. Cave’s 1986 album with The Bad Seeds, ‘Kicking Against the Pricks’, is a collection of cover versions that spans American folk idioms (‘Black Betty’, ‘Hey Joe’, ‘The Singer’), Tin-Pan-Alley balladeering (‘Something’s Gotten Hold of my Heart’, ‘By t...

  12. Nicked apomyoglobin: a noncovalent complex of two polypeptide fragments comprising the entire protein chain.

    Science.gov (United States)

    Musi, Valeria; Spolaore, Barbara; Picotti, Paola; Zambonin, Marcello; De Filippis, Vincenzo; Fontana, Angelo

    2004-05-25

    Limited proteolysis of the 153-residue chain of horse apomyoglobin (apoMb) by thermolysin results in the selective cleavage of the peptide bond Pro88-Leu89. The N-terminal (residues 1-88) and C-terminal (residues 89-153) fragments of apoMb were isolated to homogeneity and their conformational and association properties investigated in detail. Far-UV circular dichroism (CD) measurements revealed that both fragments in isolation acquire a high content of helical secondary structure, while near-UV CD indicated the absence of tertiary structure. A 1:1 mixture of the fragments leads to a tight noncovalent protein complex (1-88/89-153, nicked apoMb), characterized by secondary and tertiary structures similar to those of intact apoMb. The apoMb complex binds heme in a nativelike manner, as given by CD measurements in the Soret region. Second-derivative absorption spectra in the 250-300 nm region provided evidence that the degree of exposure of Tyr residues in the nicked species is similar to that of the intact protein at neutral pH. Also, the microenvironment of Trp residues, located in positions 7 and 14 of the 153-residue chain of the protein, is similar in both protein species, as given by fluorescence emission data. Moreover, in analogy to intact apoMb, the nicked protein binds the hydrophobic dye 1-anilinonaphthalene-8-sulfonate (ANS). Taken together, our results indicate that the two proteolytic fragments 1-88 and 89-153 of apoMb adopt partly folded states characterized by sufficiently nativelike conformational features that promote their specific association and mutual stabilization into a nicked protein species much resembling in its structural features intact apoMb. It is suggested that the formation of a noncovalent complex upon fragment complementation can mimic the protein folding process of the entire protein chain, with the difference that the folding of the complementary fragments is an intermolecular process. In particular, this study emphasizes the

  13. Preparation and Characterization of Nicke-iron Alloy Film as Freestanding Electrode for Oxygen Evolution Reaction

    Directory of Open Access Journals (Sweden)

    Yao Mengqi

    2018-01-01

    Full Text Available This work reports the porous nicke-iron alloy film supported on stainless steel mesh as freestanding electrode for enhanced oxygen evolution reaction (OER catalyst prepared from an one step electrodeposition method. Results indicated that the porous nickle-iron alloy film exhibits a low overpotential of 270 mV at 10 mA cm-2 and excellent electroconductibility. The superior OER properties can be attributed to its novel synthetic process, conductive substrate and porous structure. This work will provide a new strategy to fabricate alloy film for OER electrocatalyst.

  14. Dehalogenation of chloroalkanes by nickel(i) porphyrin derivatives, a computational study.

    Science.gov (United States)

    Szatkowski, L; Hall, M B

    2016-11-14

    The nickel(i) octaethylisobacteriochlorin anion ([OEiBCh-Ni (I) ] - ) is commonly used as a synthetic model of cofactor F 430 from Methyl-Coenzyme M Reductase. In this regard, experimental studies show that [OEiBCh-Ni (I) ] - can catalyze dehalogenation of aliphatic halides in DMF solution by a highly efficient S N 2 reaction. To better understand this process, we constructed theoretical models of the dehalogenation of chloromethane by a simple nickel(i) isobacteriochlorin anion and compared its reactivity with that of similar Ni (I) complexes with other porphyrin-derived ligands: porphyrin, chlorin, bactreriochlorin, hexahydroporphyrin and octahydroporphyrin. Our calculations predict that all of the porphyrin derivative's model reactions proceed through low-spin complexes. Relative to the energy of the separate reactants the theoretical activation energies (free-energy barriers with solvation corrections) for the dehalogenation of chloromethane are similar for all of the porphyrin derivatives and range for the different functionals from 10-15 kcal mol -1 for B3LYP to 5-10 kcal mol -1 for M06-L and to 13-18 kcal mol -1 for ωB97X-D. The relative free energies of the products of the dehalogenation step, L-Ni-Me adducts, have a range from -5 to -40 kcal mol -1 for all functionals; generally becoming more negative with increasing saturation of the porphyrin ligand. Moreover, no significant differences in the theoretical chlorine kinetic isotope effect were discernable with change of porphyrin ligand.

  15. Isothermal amplification detection of nucleic acids by a double-nicked beacon.

    Science.gov (United States)

    Shi, Chao; Zhou, Meiling; Pan, Mei; Zhong, Guilin; Ma, Cuiping

    2016-03-01

    Isothermal and rapid amplification detection of nucleic acids is an important technology in environmental monitoring, foodborne pathogen detection, and point-of-care clinical diagnostics. Here we have developed a novel method of isothermal signal amplification for single-stranded DNA (ssDNA) detection. The ssDNA target could be used as an initiator, coupled with a double-nicked molecular beacon, to originate amplification cycles, achieving cascade signal amplification. In addition, the method showed good specificity and strong anti-jamming capability. Overall, it is a one-pot and isothermal strand displacement amplification method without the requirement of a stepwise procedure, which greatly simplifies the experimental procedure and decreases the probability of contamination of samples. With its advantages, the method would be very useful to detect nucleic acids in point-of-care or field use. Copyright © 2015 Elsevier Inc. All rights reserved.

  16. W. Horsley Gantt, Nick, and the Pavlovian Science at Phipps Clinic.

    Science.gov (United States)

    Ruiz, Gabriel; Sánchez, Natividad

    2016-10-24

    William Horsley Gantt is well known as one to the principal proponents of Pavlovian methodology in the U.S. After a long stay at Ivan Petrovich Pavlov's laboratory in Leningrad from 1925 to 1929, Gantt was invited by Adolf Meyer to join the Henry Phipps Psychiatric Clinic, where he founded and directed the Pavlovian Laboratory from 1930 to 1964. Soon after his arrival at Phipps Clinic in 1931, Gantt began a Pavlovian research program that included the investigation of nervous disturbances in dogs and clinical researches with psychiatric patients. In these studies, Gantt combined a physiological method (the conditional reflexes approach), with a psychiatric problem (nervous disorders) in the context of Meyer's psychobiology that established the person or individual as unit of analysis. This fact, concentrating upon a single individual, made Gantt studies with dogs recognizable and interesting to physicians, psychologists, and psychiatrists who also worked on individuals. In this paper, we use archival materials -including correspondence, notebooks, and unpublished autobiographical material- to present a case study, that of William Horsley Gantt and his dog Nick. We will explore the reasons why Gantt' studies on nervous disturbances with this dog captured the interest of psychiatrists and clinical psychologists.

  17. Processive nicking activity of T4 endonuclease V on UV-irradiated chromatin

    International Nuclear Information System (INIS)

    Gruskin, E.A.; Lloyd, R.S.

    1986-01-01

    T4 endonuclease V initiates the excision repair of pyrimidine dimers in UV-irradiated T4 infected E. coli cells. The pyrimidine dimer specific nicking activity of T4 endonuclease V functions by a processive scanning on UV-irradiated DNA. Previously it has been demonstrated that introduction of endonuclease V into repair-deficient human cells causes a restoration of UV survival in these cells. This demonstrates that endonuclease V is competent to incise mammalian DNA at the site of pyrimidine dimers. In order to assess the ability of endonuclease V to act processively on DNA associated as chromatin, minichromosomes were prepared for use as a substrate. Form I DNA was reconstituted with H3, H4 +/- H1 histones by sequential dialysis steps from 2.0 M NaCl to 50 mM NaCl. Time course reactions were performed with minichromosomes containing 10 and 25 dimers per molecule. In each case the rate of disappearance of form I DNA which was associated as chromatin was decreased relative to that of naked form I DNA. Concurrent with that observation, the rate and extent of appearance of form III DNA was increased with the DNA in minichromosomes relative to naked DNA. This is diagnostic of an enhancement of processivity. The inclusion of H1 in the minichromosomes resulted in a slight additional increase in processivity relative to minichromosomes consisting only of H3 and H4

  18. An aptasensor for staphylococcus aureus based on nicking enzyme amplification reaction and rolling circle amplification.

    Science.gov (United States)

    Xu, Jingguo; Guo, Jia; Maina, Sarah Wanjiku; Yang, Yumeng; Hu, Yimin; Li, Xuanxuan; Qiu, Jiarong; Xin, Zhihong

    2018-05-15

    An ultra-sensitive aptamer-based biosensor for the detection of staphylococcus aureus was established by adopting the nicking enzyme amplification reaction (NEAR) and the rolling circle amplification (RCA) technologies. Aptamer-probe (AP), containing an aptamer and a probe sequence, was developed to act as the recognition unit of the biosensor, which was specifically bound to S. aureus. The probe was released from AP and initiated into the subsequent DNA amplification reactions where S. aureus was present, converting the detection of S. aureus to the investigation of probe oligonucleotide. The RCA amplification products contained a G-quadruplex motif and formed a three dimensional structure in presence of hemin. The G4/hemin complex showed horseradish peroxidase (HRP)-mimic activity and catalyzed the chemiluminescence reaction of luminol mediated by H 2 O 2 . The results showed that the established biosensor could detect S. aureus specifically with a good linear correlation at 5-10 4  CFU/mL. The signal values based on NEAR-RCA two-step cycle were boosted acutely, much higher than that relied on one-cycle magnification. The limit of detection (LoD) was determined to be as low as 5 CFU/mL. The established aptasensor exhibited a good discrimination of living against dead S. aureus, and can be applied to detect S. aureus in the food industry. Copyright © 2018 Elsevier Inc. All rights reserved.

  19. Last stop on the road to repair: structure of E. coli DNA ligase bound to nicked DNA-adenylate.

    Science.gov (United States)

    Nandakumar, Jayakrishnan; Nair, Pravin A; Shuman, Stewart

    2007-04-27

    NAD(+)-dependent DNA ligases (LigA) are ubiquitous in bacteria and essential for growth. Their distinctive substrate specificity and domain organization vis-a-vis human ATP-dependent ligases make them outstanding targets for anti-infective drug discovery. We report here the 2.3 A crystal structure of Escherichia coli LigA bound to an adenylylated nick, which captures LigA in a state poised for strand closure and reveals the basis for nick recognition. LigA envelopes the DNA within a protein clamp. Large protein domain movements and remodeling of the active site orchestrate progression through the three chemical steps of the ligation reaction. The structure inspires a strategy for inhibitor design.

  20. Rhombic-Shaped Nanostructures and Mechanical Properties of 2D DNA Origami Constructed with Different Crossover/Nick Designs.

    Science.gov (United States)

    Ma, Zhipeng; Huang, Yunfei; Park, Seongsu; Kawai, Kentaro; Kim, Do-Nyun; Hirai, Yoshikazu; Tsuchiya, Toshiyuki; Yamada, Hirofumi; Tabata, Osamu

    2018-01-01

    DNA origami methods enable the fabrication of various nanostructures and nanodevices, but their effective use depends on an understanding of their structural and mechanical properties and the effects of basic structural features. Frequency-modulation atomic force microscopy is introduced to directly characterize, in aqueous solution, the crossover regions of sets of 2D DNA origami based on different crossover/nick designs. Rhombic-shaped nanostructures formed under the influence of flexible crossovers placed between DNA helices are observed in DNA origami incorporating crossovers every 3, 4, or 6 DNA turns. The bending rigidity of crossovers is determined to be only one-third of that of the DNA helix, based on interhelical electrostatic forces reported elsewhere, and the measured pitches of the 3-turn crossover design rhombic-shaped nanostructures undergoing negligible bending. To evaluate the robustness of their structural integrity, they are intentionally and simultaneously stressed using force-controlled atomic force microscopy. DNA crossovers are verified to have a stabilizing effect on the structural robustness, while the nicks have an opposite effect. The structural and mechanical properties of DNA origami and the effects of crossovers and nicks revealed in this paper can provide information essential for the design of versatile DNA origami structures that exhibit specified and desirable properties. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  1. Optimization of a DNA Nicking Assay to Evaluate Oenocarpus bataua and Camellia sinensis Antioxidant Capacity

    Directory of Open Access Journals (Sweden)

    Louis-Jérôme Leba

    2014-10-01

    Full Text Available This study was aimed at assessing the DNA damage protective activity of different types of extracts (aqueous, methanolic and acetonic using an in vitro DNA nicking assay. Several parameters were optimized using the pUC18 plasmid, especially FeSO4, EDTA, solvent concentrations and incubation time. Special attention has been paid to removing the protective and damaging effect of the solvent and FeSO4 respectively, as well as to identifying the relevant positive and negative controls. For each solvent, the optimal conditions were determined: (i for aqueous extracts, 0.33 mM of FeSO4 and 0.62 mM of EDTA were incubated for 20 min at 37 °C; (ii for acetone extracts, 1.16% solvent were incubated for 15 min at 37 °C with 1.3 mM of FeSO4 and 2.5 mM of EDTA and (iii for methanol extracts, 0.16% solvent, were incubated for 1.5 h at 37 °C with 0.33 mM of FeSO4 and 0.62 mM of EDTA. Using the optimized conditions, the DNA damage protective activity of aqueous, methanolic and acetonic extracts of an Amazonian palm berry (Oenocarpus bataua and green tea (Camellia sinensis was assessed. Aqueous and acetonic Oenocarpus bataua extracts were protective against DNA damage, whereas aqueous, methanolic and acetonic extracts of Camellia sinensis extracts induced DNA damage.

  2. Genome Editing in Mouse Spermatogonial Stem Cell Lines Using TALEN and Double-Nicking CRISPR/Cas9

    Directory of Open Access Journals (Sweden)

    Takuya Sato

    2015-07-01

    Full Text Available Mouse spermatogonial stem cells (SSCs can be cultured for multiplication and maintained for long periods while preserving their spermatogenic ability. Although the cultured SSCs, named germline stem (GS cells, are targets of genome modification, this process remains technically difficult. In the present study, we tested TALEN and double-nicking CRISPR/Cas9 on GS cells, targeting Rosa26 and Stra8 loci as representative genes dispensable and indispensable in spermatogenesis, respectively. Harvested GS cell colonies showed a high targeting efficiency with both TALEN and CRISPR/Cas9. The Rosa26-targeted GS cells differentiated into fertility-competent sperm following transplantation. On the other hand, Stra8-targeted GS cells showed defective spermatogenesis following transplantation, confirming its prime role in the initiation of meiosis. TALEN and CRISPR/Cas9, when applied in GS cells, will be valuable tools in the study of spermatogenesis and for revealing the genetic mechanism of spermatogenic failure.

  3. The DNA Repair Repertoire of Mycobacterium smegmatis FenA Includes the Incision of DNA 5' Flaps and the Removal of 5' Adenylylated Products of Aborted Nick Ligation.

    Science.gov (United States)

    Uson, Maria Loressa; Ghosh, Shreya; Shuman, Stewart

    2017-09-01

    We characterize Mycobacterium smegmatis FenA as a manganese-dependent 5'-flap endonuclease homologous to the 5'-exonuclease of DNA polymerase I. FenA incises a nicked 5' flap between the first and second nucleotides of the duplex segment to yield a 1-nucleotide gapped DNA, which is then further resected in dinucleotide steps. Initial FenA cleavage at a Y-flap or nick occurs between the first and second nucleotides of the duplex. However, when the template 3' single strand is eliminated to create a 5'-tailed duplex, FenA incision shifts to between the second and third nucleotides. A double-flap substrate with a mobile junction (mimicking limited strand displacement synthesis during gap repair) is preferentially incised as the 1-nucleotide 3'-flap isomer, with the scissile phosphodiester shifted by one nucleotide versus a static double flap. FenA efficiently removes the 5' App(dN) terminus of an aborted nick ligation reaction intermediate, thereby highlighting FenA as an agent of repair of such lesions, which are formed under a variety of circumstances by bacterial NAD + -dependent DNA ligases and especially by mycobacterial DNA ligases D and C. IMPORTANCE Structure-specific DNA endonucleases are implicated in bacterial DNA replication, repair, and recombination, yet there is scant knowledge of the roster and catalytic repertoire of such nucleases in Mycobacteria This study identifies M. smegmatis FenA as a stand-alone endonuclease homologous to the 5'-exonuclease domain of mycobacterial DNA polymerase 1. FenA incises 5' flaps, 5' nicks, and 5' App(dN) intermediates of aborted nick ligation. The isolated N-terminal domain of M. smegmatis Pol1 is also shown to be a flap endonuclease. Copyright © 2017 American Society for Microbiology.

  4. Konstrukce vřeten vícevřetenového soustružnického automatu

    OpenAIRE

    Kráčmar, Tomáš

    2016-01-01

    Obsahem diplomové práce je konstrukce vřeten vícevřetenového soustružnického automatu pro práci z tyče maximálního průměru 7 mm. Práce se zabývá novou koncepcí pohonu vřeten, kde jsou vřetena poháněna externími asynchronními motory přes ozubená kola s vnitřním ozubením uložená vně vřetenového bubnu namísto současného způsobu pohonu centrálními koaxiálními hřídeli. Součástí práce je rešerše vícevřetenových automatů včetně popisu hlavních uzlů, konstrukční návrh, pevnostní výpočty, výpočty trva...

  5. DNA gyrase with a single catalytic tyrosine can catalyze DNA supercoiling by a nicking-closing mechanism

    Science.gov (United States)

    Gubaev, Airat; Weidlich, Daniela; Klostermeier, Dagmar

    2016-01-01

    The topological state of DNA is important for replication, recombination and transcription, and is regulated in vivo by DNA topoisomerases. Gyrase introduces negative supercoils into DNA at the expense of ATP hydrolysis. It is the accepted view that gyrase achieves supercoiling by a strand passage mechanism, in which double-stranded DNA is cleaved, and a second double-stranded segment is passed through the gap, converting a positive DNA node into a negative node. We show here that gyrase with only one catalytic tyrosine that cleaves a single strand of its DNA substrate can catalyze DNA supercoiling without strand passage. We propose an alternative mechanism for DNA supercoiling via nicking and closing of DNA that involves trapping, segregation and relaxation of two positive supercoils. In contrast to DNA supercoiling, ATP-dependent relaxation and decatenation of DNA by gyrase lacking the C-terminal domains require both tyrosines and strand passage. Our results point towards mechanistic plasticity of gyrase and might pave the way for finding novel and specific mechanism-based gyrase inhibitors. PMID:27557712

  6. On the use of an appropriate TdT-mediated dUTP-biotin nick end labeling assay to identify apoptotic cells.

    OpenAIRE

    Lebon, C.; Rodriguez, G.V.; Zaoui, I.E.; Jaadane, I.; Behar-Cohen, F.; Torriglia, A.

    2015-01-01

    Apoptosis is an essential cellular mechanism involved in many processes such as embryogenesis, metamorphosis, and tissue homeostasis. DNA fragmentation is one of the key markers of this form of cell death. DNA fragmentation is executed by endogenous endonucleases such as caspase-activated DNase (CAD) in caspase-dependent apoptosis. The TUNEL (TdT-mediated dUTP-biotin nick end labeling) technique is the most widely used method to identify apoptotic cells in a tissue or culture and to assess dr...

  7. DNA scanning mechanism of T4 endonuclease V. Effect of NaCl concentration on processive nicking activity

    International Nuclear Information System (INIS)

    Gruskin, E.A.; Lloyd, R.S.

    1986-01-01

    T4 endonuclease V is a pyrimidine dimer-specific endonuclease which generates incisions in DNA at the sites of pyrimidine dimers by a processive reaction mechanism. A model is presented in which the degree of processivity is directly related to the efficacy of the one-dimensional diffusion of endonuclease V on DNA by which the enzyme locates pyrimidine dimers. The modulation of the processive nicking activity of T4 endonuclease V on superhelical covalently closed circular DNA (form I) which contains pyrimidine dimers has been investigated as a function of the ionic strength of the reaction. Agarose gel electrophoresis was used to separate the three topological forms of the DNA which were generated in time course reactions of endonuclease V with dimer-containing form I DNA in the absence of NaCl, and in 25, 50, and 100 mM NaCl. The degree of processivity was evaluated in terms of the mass fraction of form III (linear) DNA which was produced as a function of the fraction of form I DNA remaining. Processivity is maximal in the absence of NaCl and decreases as the NaCl concentration is increased. At 100 mM NaCl, processivity is abolished and endonuclease V generates incisions in DNA at the site of dimers by a distributive reaction mechanism. The change from the distributive to a processive reaction mechanism occurs at NaCl concentrations slightly below 50 mM. The high degree of processivity which is observed in the absence of NaCl is reversible to the distributive mechanism, as demonstrated by experiments in which the NaCl concentration was increased during the time course reaction. In addition, unirradiated DNA inhibited the incision of irradiated DNA only at NaCl concentrations at which processivity was observed

  8. Book Reviews / Nick Ellison

    Index Scriptorium Estoniae

    Ellison, Nick

    2008-01-01

    J. Aidukaite doktoridissertatsiooni, The Emergence of the Post-Socialist Welfare State - The Case of the Baltic States: Estonia, Latvia and Lithuania, milles võrreldakse nende riikide sotsiaalpoliitikat ja sotsiaalabi süsteeme, refereering

  9. Repair of pyrimidine dimers in radiation-sensitive mutants rad3, rad4, rad6, and rad9 of Saccharomyces cerevisiae. [nicking

    Energy Technology Data Exchange (ETDEWEB)

    Prakash, L [Rochester Univ., N.Y. (USA). Dept. of Radiation Biology and Biophysics; Rochester Univ., N.Y. (USA). School of Medicine and Dentistry)

    1977-10-01

    The ability to remove ultraviolet-induced pyrimidine dimers was examined in four radiation-sensitive mutants of Saccharomyces cerevisiae. The susceptibility of DNA from irradiated cells to nicking by either the T4 uv-endonuclease or an endonuclease activity found in crude extracts of Micrococcus luteus was used to measure the presence of dimers in DNA. The rad3 and rad4 mutants are shown to be defective in dimer excision whereas the rad6 and rad9 mutants are proficient in dimer excision.

  10. Site- and strand-specific nicking of DNA by fusion proteins derived from MutH and I-SceI or TALE repeats.

    Science.gov (United States)

    Gabsalilow, Lilia; Schierling, Benno; Friedhoff, Peter; Pingoud, Alfred; Wende, Wolfgang

    2013-04-01

    Targeted genome engineering requires nucleases that introduce a highly specific double-strand break in the genome that is either processed by homology-directed repair in the presence of a homologous repair template or by non-homologous end-joining (NHEJ) that usually results in insertions or deletions. The error-prone NHEJ can be efficiently suppressed by 'nickases' that produce a single-strand break rather than a double-strand break. Highly specific nickases have been produced by engineering of homing endonucleases and more recently by modifying zinc finger nucleases (ZFNs) composed of a zinc finger array and the catalytic domain of the restriction endonuclease FokI. These ZF-nickases work as heterodimers in which one subunit has a catalytically inactive FokI domain. We present two different approaches to engineer highly specific nickases; both rely on the sequence-specific nicking activity of the DNA mismatch repair endonuclease MutH which we fused to a DNA-binding module, either a catalytically inactive variant of the homing endonuclease I-SceI or the DNA-binding domain of the TALE protein AvrBs4. The fusion proteins nick strand specifically a bipartite recognition sequence consisting of the MutH and the I-SceI or TALE recognition sequences, respectively, with a more than 1000-fold preference over a stand-alone MutH site. TALE-MutH is a programmable nickase.

  11. inflammatory investigations of stem bark of Daniellia oliveri

    African Journals Online (AJOL)

    GRACE

    2006-05-16

    May 16, 2006 ... 3Department of Pharmacognosy and Drug Development, Ahmadu Bello University, Zaria, NIGERIA. ... inflammatory effects in mice and rats, respectively. ..... McGraw-Hill (Health Profession Division), New. Page 6. York. pp. 63 – 65. Dalziel JM (1937). Ceasalpiniaceae The useful plants of West Tropical.

  12. Electric Six : Ettevaatust! Kõrgepinge!

    Index Scriptorium Estoniae

    2007-01-01

    Ameerika rockansamblist Electric Six, albumitest "Fire", "Swizerland". Ameerika rockauljast-kitarristist Nick Oliveri'st, ansambli Mondo Generator (www.mondogenerator.com) albumist "Dead Planet: SonicSlowMotionTrails". Info festivalist: www.rabarock.delfi.ee / www.rabarock.ee

  13. Ultrasensitive electrochemical biosensor for detection of DNA from Bacillus subtilis by coupling target-induced strand displacement and nicking endonuclease signal amplification.

    Science.gov (United States)

    Hu, Yuhua; Xu, Xueqin; Liu, Qionghua; Wang, Ling; Lin, Zhenyu; Chen, Guonan

    2014-09-02

    A simple, ultrasensitive, and specific electrochemical biosensor was designed to determine the given DNA sequence of Bacillus subtilis by coupling target-induced strand displacement and nicking endonuclease signal amplification. The target DNA (TD, the DNA sequence from the hypervarient region of 16S rDNA of Bacillus subtilis) could be detected by the differential pulse voltammetry (DPV) in a range from 0.1 fM to 20 fM with the detection limit down to 0.08 fM at the 3s(blank) level. This electrochemical biosensor exhibits high distinction ability to single-base mismatch, double-bases mismatch, and noncomplementary DNA sequence, which may be expected to detect single-base mismatch and single nucleotide polymorphisms (SNPs). Moreover, the applicability of the designed biosensor for detecting the given DNA sequence from Bacillus subtilis was investigated. The result obtained by electrochemical method is approximately consistent with that by a real-time quantitative polymerase chain reaction detecting system (QPCR) with SYBR Green.

  14. Nicking endonuclease-assisted signal amplification of a split molecular aptamer beacon for biomolecule detection using graphene oxide as a sensing platform.

    Science.gov (United States)

    Li, Xiang; Ding, Xuelian; Fan, Jing

    2015-12-07

    Sensitive and selective detection of ultralow concentrations of specific biomolecules is important in early clinical diagnoses and biomedical applications. Many types of aptasensors have been developed for the detection of various biomolecules, but usually suffer from false positive signals and high background signals. In this work, we have developed an amplified fluorescence aptasensor platform for ultrasensitive biomolecule detection based on enzyme-assisted target-recycling signal amplification and graphene oxide. By using a split molecular aptamer beacon and a nicking enzyme, the typical problem of false positive signals can be effectively resolved. Only in the presence of a target biomolecule, the sensor system is able to generate a positive signal, which significantly improves the selectivity of the aptasensor. Moreover, using graphene oxide as a super-quencher can effectively reduce the high background signal of a sensing platform. We select vascular endothelial growth factor (VEGF) and adenosine triphosphate (ATP) as model analytes in the current proof-of-concept experiments. It is shown that under optimized conditions, our strategy exhibits high sensitivity and selectivity for the quantification of VEGF and ATP with a low detection limit (1 pM and 4 nM, respectively). In addition, this biosensor has been successfully utilized in the analysis of real biological samples.

  15. Once a physicist: Nick Horvath

    Science.gov (United States)

    Horvath, Nick

    2009-05-01

    How did you get into basketball? That's an easy one: I was tall. I had zero interest in sport at a young age and was pushed into basketball against my will when I was about eight. I hated it. The next year, however, I was so much taller than everyone else that in my first game, even though I had no idea what I was doing, I could simply hold the ball above my head and shoot over everyone. I fell in love with basketball after that first game. If I had played badly that day, it is likely that I would not have played again in my entire life.

  16. Analysis of Apoptosis in Ultraviolet-Induced Sea Cucumber (Stichopus japonicus) Melting Using Terminal Deoxynucleotidyl-Transferase-Mediated dUTP Nick End-Labeling Assay and Cleaved Caspase-3 Immunohistochemistry.

    Science.gov (United States)

    Yang, Jing-Feng; Gao, Rong-Chun; Wu, Hai-Tao; Li, Peng-Fei; Hu, Xian-Shu; Zhou, Da-Yong; Zhu, Bei-Wei; Su, Yi-Cheng

    2015-11-04

    The sea cucumber body wall melting phenomenon occurs under certain circumstances, and the mechanism of this phenomenon remains unclear. This study investigated the apoptosis in the ultraviolet (UV)-induced sea cucumber melting phenomenon. Fresh sea cucumbers (Stichopus japonicus) were exposed to UV radiation for half an hour at an intensity of 0.056 mW/cm(2) and then held at room temperature for melting development. The samples were histologically processed into formalin-fixed paraffin-embedded tissues. The apoptosis of samples was analyzed with the terminal deoxynucleotidyl-transferase-mediated dUTP nick end-labeling (TUNEL) assay and cleaved caspase-3 immunohistochemistry. The emergence of TUNEL-positive cells speeds up between 0.5 and 2 h after UV irradiation. Cleaved caspase-3 positive cells were obviously detected in sample tissues immediately after the UV irradiation. These results demonstrated that sea cucumber melting induced by UV irradiation was triggered by the activation of caspase-3 followed by DNA fragmentation in sea cucumber tissue, which was attributed to apoptosis but was not a consequence of autolysis activity.

  17. C-terminal phenylalanine of bacteriophage T7 single-stranded DNA-binding protein is essential for strand displacement synthesis by T7 DNA polymerase at a nick in DNA.

    Science.gov (United States)

    Ghosh, Sharmistha; Marintcheva, Boriana; Takahashi, Masateru; Richardson, Charles C

    2009-10-30

    Single-stranded DNA-binding protein (gp2.5), encoded by gene 2.5 of bacteriophage T7, plays an essential role in DNA replication. Not only does it remove impediments of secondary structure in the DNA, it also modulates the activities of the other replication proteins. The acidic C-terminal tail of gp2.5, bearing a C-terminal phenylalanine, physically and functionally interacts with the helicase and DNA polymerase. Deletion of the phenylalanine or substitution with a nonaromatic amino acid gives rise to a dominant lethal phenotype, and the altered gp2.5 has reduced affinity for T7 DNA polymerase. Suppressors of the dominant lethal phenotype have led to the identification of mutations in gene 5 that encodes the T7 DNA polymerase. The altered residues in the polymerase are solvent-exposed and lie in regions that are adjacent to the bound DNA. gp2.5 lacking the C-terminal phenylalanine has a lower affinity for gp5-thioredoxin relative to the wild-type gp2.5, and this affinity is partially restored by the suppressor mutations in DNA polymerase. gp2.5 enables T7 DNA polymerase to catalyze strand displacement DNA synthesis at a nick in DNA. The resulting 5'-single-stranded DNA tail provides a loading site for T7 DNA helicase. gp2.5 lacking the C-terminal phenylalanine does not support this event with wild-type DNA polymerase but does to a limited extent with T7 DNA polymerase harboring the suppressor mutations.

  18. C-terminal Phenylalanine of Bacteriophage T7 Single-stranded DNA-binding Protein Is Essential for Strand Displacement Synthesis by T7 DNA Polymerase at a Nick in DNA*

    Science.gov (United States)

    Ghosh, Sharmistha; Marintcheva, Boriana; Takahashi, Masateru; Richardson, Charles C.

    2009-01-01

    Single-stranded DNA-binding protein (gp2.5), encoded by gene 2.5 of bacteriophage T7, plays an essential role in DNA replication. Not only does it remove impediments of secondary structure in the DNA, it also modulates the activities of the other replication proteins. The acidic C-terminal tail of gp2.5, bearing a C-terminal phenylalanine, physically and functionally interacts with the helicase and DNA polymerase. Deletion of the phenylalanine or substitution with a nonaromatic amino acid gives rise to a dominant lethal phenotype, and the altered gp2.5 has reduced affinity for T7 DNA polymerase. Suppressors of the dominant lethal phenotype have led to the identification of mutations in gene 5 that encodes the T7 DNA polymerase. The altered residues in the polymerase are solvent-exposed and lie in regions that are adjacent to the bound DNA. gp2.5 lacking the C-terminal phenylalanine has a lower affinity for gp5-thioredoxin relative to the wild-type gp2.5, and this affinity is partially restored by the suppressor mutations in DNA polymerase. gp2.5 enables T7 DNA polymerase to catalyze strand displacement DNA synthesis at a nick in DNA. The resulting 5′-single-stranded DNA tail provides a loading site for T7 DNA helicase. gp2.5 lacking the C-terminal phenylalanine does not support this event with wild-type DNA polymerase but does to a limited extent with T7 DNA polymerase harboring the suppressor mutations. PMID:19726688

  19. Anti-Diarrheal Activity Of The Leaf Extracts Of Daniellia oliveri Hutch ...

    African Journals Online (AJOL)

    Moraceae) used in diarrhea treatment in Hausa ethnomedicine of Northern Nigeria were investigated. The study was carried out on parfused isolated rabbit jejunum and castor oil-induced diarrhea in mice. The n-butanol extracts: NBD and NBF ...

  20. The Portraiture of Nick: Scene Two, the Middle Years

    Science.gov (United States)

    Bicehouse, Vaughn L.

    2012-01-01

    In 2007, the Autism Society of America (ASA) estimated that 1.5 million Americans and their families were affected by autism. As the current Autistic Spectrum Disorder (ASD) prevalence rates continue to rise (10%-17% each year) so does the awareness that ASD is no respecter of persons. ASD touches children of every racial, ethnic, and…

  1. Greens protest at U. S. Embassy / Nick Coleman

    Index Scriptorium Estoniae

    Coleman, Nick

    2000-01-01

    Protestijad kogunesid USA Riia saatkonna hoone ette. Väljendati oma rahulolematust USA tahtmatuse suhtes vähendada süsinikdioksüüdi paiskamist õhkkonda, mis ilmnes eelmisel kuul peetud kliimamuutuse konverentsil Haagis

  2. Ten years of transition and... / Rika Ishii, Nick Sten

    Index Scriptorium Estoniae

    Ishii, Rika

    1999-01-01

    Ülevaade majandusreformidest Baltimaades ja Baltimaade majandussuhetest Euroopa Liidu ning Venemaaga. Tabel: SKP, inflatsioon, väliskaubandus ja välisinvesteeringud Eestis, Lätis, Leedus ja Kesk- ja Ida-Euroopa riikides aastatel 1991-1998

  3. In the Nick of Time: Proactive Prevention of Obligation Violations

    DEFF Research Database (Denmark)

    Basin, David; Debois, Søren; Hildebrandt, Thomas

    2016-01-01

    We present a system model, an enforcement mechanism, and a policy language for the proactive enforcement of timed provisions and obligations. Our approach improves upon existing formalisms in two ways: (1) we exploit the target system's existing functionality to avert policy violations proactively...... declaratively express timed provisions and obligations as causal relationships between events, and DCR states explicitly represent pending obligations. As key technical results, we show that enforceability of DCR policies is decidable, we give a sufficient polynomial time verifiable condition for a policy...... to be enforceable, and we give an algorithm for determining from a DCR state a sequence of actions that discharge impending obligations....

  4. Review Article: The Vanished Ghosts in Two Hungarian Family Memoirs. Farkas, Charles. 2013. Vanished by the Danube: Peace, War, Revolution, and Flight to the West (Introduction by Margaret McMullan. Albany: Excelsior Editions, State University of New York Press. 472 pp. Illus; and Barlay, Nick. 2013. Scattered Ghosts: One Family's Survival through War, Holocaust and Revolution. London and New York: I. B. Tauris. 240 pp. Illus.

    Directory of Open Access Journals (Sweden)

    Ruth G. Biro

    2015-01-01

    Full Text Available Recent personal documentary works about major historical events of the twentieth century, e.g., World War II, the Holocaust and the 1956 Hungarian Revolution, offer their readers a rich and multifaceted narrative, or a history that is also "his story," "her story" and that of entire families, cohorts and communities. Often, these works are accompanied by visual artifacts such as photographs, family tress, maps etc., or supported by concise historical surveys. Thus these memoirs complete the work of historians with the lived experiences of the few that represent many. Such is the case with two 2013 books by Charles Farkas and Nick Barlay depicting their mid-twentieth century Hungarian families, one Christian and one Jewish, through two World Wars and the anti-communist uprising, culminating in their escape to the West and in the two authors looking back upon the Hungarian past of their families.

  5. Processing of Nonconjugative Resistance Plasmids by Conjugation Nicking Enzyme of Staphylococci

    Energy Technology Data Exchange (ETDEWEB)

    Pollet, Rebecca M.; Ingle, James D.; Hymes, Jeff P.; Eakes, Thomas C.; Eto, Karina Yui; Kwong, Stephen M.; Ramsay, Joshua P.; Firth, Neville; Redinbo, Matthew R. (Curtin U.); (Sydney); (UNC)

    2016-01-04

    Antimicrobial resistance inStaphylococcus aureuspresents an increasing threat to human health. This resistance is often encoded on mobile plasmids, such as pSK41; however, the mechanism of transfer of these plasmids is not well understood. In this study, we first examine key protein-DNA interactions formed by the relaxase enzyme, NES, which initiates and terminates the transfer of the multidrug resistance plasmid pSK41. Two loops on the NES protein, hairpin loops 1 and 2, form extensive contacts with the DNA hairpin formed at theoriTregion of pSK41, and here we establish that these contacts are essential for proper DNA cleavage and religation by the full 665-residue NES proteinin vitro. Second, pSK156 and pCA347 are nonconjugativeStaphylococcus aureusplasmids that contain sequences similar to theoriTregion of pSK41 but differ in the sequence predicted to form a DNA hairpin. We show that pSK41-encoded NES is able to bind, cleave, and religate theoriTsequences of these nonconjugative plasmidsin vitro. Although pSK41 could mobilize a coresident plasmid harboring its cognateoriT, it was unable to mobilize plasmids containing the pSK156 and pCA347 variantoriTmimics, suggesting that an accessory protein like that previously shown to confer specificity in the pWBG749 system may also be involved in transmission of plasmids containing a pSK41-likeoriT. These data indicate that the conjugative relaxase intransmechanism recently described for the pWBG749 family of plasmids also applies to the pSK41 family of plasmids, further heightening the potential significance of this mechanism in the horizontal transfer of staphylococcal plasmids.

    IMPORTANCEUnderstanding the mechanism of antimicrobial resistance transfer in bacteria such asStaphylococcus aureusis an important step toward potentially slowing the spread of antimicrobial-resistant infections. This work establishes protein-DNA interactions essential for the transfer of theStaphylococcus aureusmultiresistance plasmid pSK41 by its relaxase, NES. This enzyme also processed variantoriT-like sequences found on numerous plasmids previously considered nontransmissible, suggesting that in conjunction with an uncharacterized accessory protein, these plasmids may be transferred horizontally via a relaxase intransmechanism. These findings have important implications for our understanding of staphylococcal resistance plasmid evolution.

  6. "Esst estnische Pasteten in estnischen Nischen" : Ilmar Laabans poetische Welt / Gisbert Jänicke

    Index Scriptorium Estoniae

    Jänicke, Gisbert, 1937-

    2000-01-01

    Artikkel sisaldab I. Laabani luuletusi saksa k.: Das Ende der Ankerkette ist des Gesanges Beginn ; Ich liebe euch ; "Die Quelle des Tods in sauberen Wolken..." ; Vögel des Selbstmords ; "Ich zerstreue mich über meinen Tisch..." ; Pornografisch ; Die Moral des Nordlichts ; Mikrokosmos 1, 20, 23, 26, 29, 30, 34, 49. Samuti sisaldab artikkel I. Laabani rootsik. luuletuse "Hur står det till?" saksak. teksti ja I. Laabani saksa k. kirjutatud luuletuse "Funkenspielstück".

  7. Výzkum diváka Strašnického divadla

    OpenAIRE

    Humlová, Jana

    2012-01-01

    This bachelor thesis focuses on marketing applied by theatres. Firstly there is a definition of marketing in general and then the thesis focuses on differences in marketing applied by cultural organizations, especially by theatres. Then it shows diverse types of theatres. Afterwards the thesis becomes less theoretical and gets more concrete and it explains the term marketing mix on example of Strašnice theatre. The practical part is represented by a marketing research. The purpose of the rese...

  8. Nick Jonas on Type 1 Diabetes | NIH MedlinePlus the Magazine

    Science.gov (United States)

    Skip to main content NIH MedlinePlus the Magazine NIH MedlinePlus Salud Download the Current Issue PDF [1.9 mb] Trusted Health Information from the National Institutes of Health Home Current Issue ...

  9. Triplex configuration in the nick-free DNAs that constitute the chromosomal scaffolds in grasshopper spermatids

    Czech Academy of Sciences Publication Activity Database

    Černá, Adriana; Lopez-Fernandez, C.; Fernandez, J.L.; de la Espina, S.M.D.; De la Torre, C.; Gosalvez, J.

    2008-01-01

    Roč. 117, č. 1 (2008), s. 15-24 ISSN 0009-5915 Institutional research plan: CEZ:AV0Z50380511 Keywords : chromatid scaffold * DNA loops * triplex DNA Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 5.111, year: 2008

  10. Im Wind / Mari Vallisoo ; tõlk. Gisbert Jänicke

    Index Scriptorium Estoniae

    Vallisoo, Mari, 1950-2013

    2003-01-01

    Sisu: Im Wind = Tuulega ; Zwei Frauen = Kaks naist ; In Gedanken an Aleksander Levin = aleksander Levinile mõteldes ; Jetzt im Dunkel = Nüüd pimedas ; Fragen = Küsimused ; Der Zwillingsbruder = Kaksikvend

  11. Zehn Oden / Kristjan Jaak Peterson ; [Deutsch von Gisbert Jänicke

    Index Scriptorium Estoniae

    Peterson, Kristian Jaak, 1801-1822

    2001-01-01

    On ilmunud ka kogumikus "Iaak. Kristian Jaak Peterson 200" (Tln. : Eesti Keele Sihtasutus, 2001, lk. 105-122). Sisu: Der Mond = Kuu ; Freundschaft = Sõprus ; Der Sänger = Laulja ; An Gott = Jumalale ; Hoffnung = Lootus ; Ich muss Trinken. Liedchen = Ma pean jooma. Lauluke ; Ich bin wieder glücklich = Olen jälle õnnis ; Der Mensch = Inimene ; Sonnenuntergang = Päeva loojaminemine ; Herbst = Sügise.

  12. Gedichte aus den Jahren 1968 bis 1975 / Viivi Luik ; tõlk. Gisbert Jänicke

    Index Scriptorium Estoniae

    Luik, Viivi, 1946-

    1999-01-01

    Tekst saksa ja eesti keeles. Sisu: Weite = Avarus ; "Wie der Krieg..." = "Otsekui sõda..." ; "es vergehn..." = "hävivad..." ; Das Geheimnis = Saladus ; "Ob auch ich weiss..." = "Kas minagi tean..." ; "Ich redete einmal von Feldern..." = "Rääkisin väljadest ükskord..." ; Andere = Teine ; Der Tag = Päev

  13. Tabellae et tabulata : [luuletused] / Hasso Krull ; aus dem Estnischen von Gisbert Jänicke

    Index Scriptorium Estoniae

    Krull, Hasso, 1964-

    2001-01-01

    Sisu : Der Kopf dreht sich [Pea käib ringi] ; Gedicht [Luuletus] ; Vier Tafeln 1-4 [Neli tabelit 1-4] ; I : "Gerade hier entlang kamen wir in den Verkehr..." [I : "Just siitkaudu tulime liikumisele lähedale..."] ; II : "Angenehmer Kaffeeduft. Von einer Stärke, die..." [II : "Meeldiv kohvilõhn. Sellel on jõudu, mis toob..."] ; III : "Hesekiel schreibt, "ihre Felgen waren so hoch, dass ich..." [III : "Hesekiel märgib, et "nende pöiad olid kõrged ning..."] ; IV : "Zwangsläufig wird der Luftdruck immer spürbarer..." [IV : "Paratamatult muutub õhurõhk üha märgatavamaks ..."] ; V : "Ich war im Wald, sammelte Blätter von den Bäumen, ass sie..." [V : "Olin metsa all, korjasin puulehti, sõin neid..."] ; VI : "Der Grund für die Gewalt ist immer der andere. Der..." ["Vägivalla alus on just üks Teine. Kes..."] ; - : "Grüsst euch. Euch auch..." [-: "No tere. Tere ka..."

  14. Zehn Oden = Kümme oodi / Kristjan Jaak Peterson ; Deutsch von Gisbert Jänicke

    Index Scriptorium Estoniae

    Peterson, Kristjan Jaak, 1801-1822

    2001-01-01

    Sisu: An Gott ; Sonnenuntergang ; Herbst ; Der Mensch ; Der Mond ; Freundschaft ; Ich muss trinken. Liedchen ; Hoffnung ; Ich bin wieder glücklich ; Der Sänger. Orig.: Jummalale ; Päva loja-minneminne ; Süggise ; Innimenne ; Kuu ; Söbradus ; Ma pean joma. Lauloke ; Lotus ; Ollen jälle önnis ; Laulja

  15. Preparation of short cytosine-modified oligonucleotides by nicking enzyme amplification reaction

    Czech Academy of Sciences Publication Activity Database

    Ménová, Petra; Hocek, Michal

    2012-01-01

    Roč. 48, č. 55 (2012), s. 6921-6923 ISSN 1359-7345 R&D Projects: GA ČR GA203/09/0317 Institutional support: RVO:61388963 Keywords : cross - coupling reactions * nucleoside triphosphates * functionalized DNA * restriction endonucleases * polymerase incorporation Subject RIV: CC - Organic Chemistry Impact factor: 6.378, year: 2012

  16. Distribuce předpon v českém sylabotónickém trocheji

    Czech Academy of Sciences Publication Activity Database

    Plecháč, Petr; Kolár, Robert; Hlaváčová, J.; Merthová, K.

    2017-01-01

    Roč. 78, č. 4 (2017), s. 322-332 ISSN 0037-7031 R&D Projects: GA ČR(CZ) GA14-31160S Institutional support: RVO:68378068 Keywords : metrics and prosody * morphology * corpus linguistics * trochee * prefixes Subject RIV: AJ - Letters, Mass-media, Audiovision OBOR OECD: Specific literatures Impact factor: 0.625, year: 2016

  17. Fluorescence Quenching in Oligonucleotides Containing 7-Substituted 7-Deazaguanine Bases Prepared by the Nicking Enzyme Amplification Reaction

    Czech Academy of Sciences Publication Activity Database

    Ménová, Petra; Dziuba, Dmytro; Güixens-Gallardo, Pedro; Jurkiewicz, Piotr; Hof, Martin; Hocek, Michal

    2015-01-01

    Roč. 26, č. 2 (2015), s. 361-366 ISSN 1043-1802 R&D Projects: GA ČR GBP206/12/G151 Institutional support: RVO:61388963 ; RVO:61388955 Keywords : ethidium bromide * electrochemical detection * DNA Subject RIV: CC - Organic Chemistry; CF - Physical ; Theoretical Chemistry (UFCH-W) Impact factor: 4.500, year: 2015

  18. Structures of minute virus of mice replication initiator protein N-terminal domain: Insights into DNA nicking and origin binding

    International Nuclear Information System (INIS)

    Tewary, Sunil K.; Liang, Lingfei; Lin, Zihan; Lynn, Annie; Cotmore, Susan F.; Tattersall, Peter; Zhao, Haiyan; Tang, Liang

    2015-01-01

    Members of the Parvoviridae family all encode a non-structural protein 1 (NS1) that directs replication of single-stranded viral DNA, packages viral DNA into capsid, and serves as a potent transcriptional activator. Here we report the X-ray structure of the minute virus of mice (MVM) NS1 N-terminal domain at 1.45 Å resolution, showing that sites for dsDNA binding, ssDNA binding and cleavage, nuclear localization, and other functions are integrated on a canonical fold of the histidine-hydrophobic-histidine superfamily of nucleases, including elements specific for this Protoparvovirus but distinct from its Bocaparvovirus or Dependoparvovirus orthologs. High resolution structural analysis reveals a nickase active site with an architecture that allows highly versatile metal ligand binding. The structures support a unified mechanism of replication origin recognition for homotelomeric and heterotelomeric parvoviruses, mediated by a basic-residue-rich hairpin and an adjacent helix in the initiator proteins and by tandem tetranucleotide motifs in the replication origins. - Highlights: • The structure of a parvovirus replication initiator protein has been determined; • The structure sheds light on mechanisms of ssDNA binding and cleavage; • The nickase active site is preconfigured for versatile metal ligand binding; • The binding site for the double-stranded replication origin DNA is identified; • A single domain integrates multiple functions in virus replication

  19. Structures of minute virus of mice replication initiator protein N-terminal domain: Insights into DNA nicking and origin binding

    Energy Technology Data Exchange (ETDEWEB)

    Tewary, Sunil K.; Liang, Lingfei; Lin, Zihan; Lynn, Annie [Department of Molecular Biosciences, University of Kansas, Lawrence, KS 66045 (United States); Cotmore, Susan F. [Departments of Laboratory Medicine, Yale University Medical School, New Haven, CT 06510 (United States); Tattersall, Peter [Departments of Laboratory Medicine, Yale University Medical School, New Haven, CT 06510 (United States); Departments of Genetics, Yale University Medical School, New Haven, CT 06510 (United States); Zhao, Haiyan, E-mail: zhaohy@ku.edu [Department of Molecular Biosciences, University of Kansas, Lawrence, KS 66045 (United States); Tang, Liang, E-mail: tangl@ku.edu [Department of Molecular Biosciences, University of Kansas, Lawrence, KS 66045 (United States)

    2015-02-15

    Members of the Parvoviridae family all encode a non-structural protein 1 (NS1) that directs replication of single-stranded viral DNA, packages viral DNA into capsid, and serves as a potent transcriptional activator. Here we report the X-ray structure of the minute virus of mice (MVM) NS1 N-terminal domain at 1.45 Å resolution, showing that sites for dsDNA binding, ssDNA binding and cleavage, nuclear localization, and other functions are integrated on a canonical fold of the histidine-hydrophobic-histidine superfamily of nucleases, including elements specific for this Protoparvovirus but distinct from its Bocaparvovirus or Dependoparvovirus orthologs. High resolution structural analysis reveals a nickase active site with an architecture that allows highly versatile metal ligand binding. The structures support a unified mechanism of replication origin recognition for homotelomeric and heterotelomeric parvoviruses, mediated by a basic-residue-rich hairpin and an adjacent helix in the initiator proteins and by tandem tetranucleotide motifs in the replication origins. - Highlights: • The structure of a parvovirus replication initiator protein has been determined; • The structure sheds light on mechanisms of ssDNA binding and cleavage; • The nickase active site is preconfigured for versatile metal ligand binding; • The binding site for the double-stranded replication origin DNA is identified; • A single domain integrates multiple functions in virus replication.

  20. Gedichte : [aus "Talv"¡ ("Winter")] : [luuletused] / Hasso Krull ; tlk. Irja Grönholm ja Gisbert Jänicke

    Index Scriptorium Estoniae

    Krull, Hasso, 1964-

    2006-01-01

    Sisu: Winter : "Die Demokratie. Sie ist in gewissem Sinn ein Kastensystem..." ; "Die Erdmännchen. Ihre Hochzeit..." ; "Die Bären. Wachen auf und fragen..." ; "Die Wanzen. Wohnten in der Matratze..." ; "Die Folter. Schon die christlich Inquisition..." ; "Das Gedächtnis. Alle schreiben sie davon..." ; "Einen Gott gibt es...". Orig.: Talv : "Demokraatia. See on omamoodi kastisüsteem..." ; "Kümnekingalised. Nende pulm..." ; "Karud. Ärkavad üles ja küsivad..." ; "Lutikad. Elasid madratsi sees..." ; "Piinamine. Juba kristlik inkvisitsioon..." ; "Mälu. Kõik kirjutavad sellest..." ; "Üks jumal on olemas..."

  1. In the Nick of Time: A Heteroskedastic SVAR Model and Its Application to the Crude Oil Futures Market

    NARCIS (Netherlands)

    Sun, Hang; Bos, Jaap W.B.; Li, Zhuo

    2017-01-01

    Many economic analyses revolve around the identification of shocks. However, this becomes difficult if we do not have enough information, for example because we do not observe the underlying process at a high enough frequency. As a result, if the response of one variable to a shock to another takes

  2. Der Meteorologe : (aus dem Band "V". Tallinn 1998) / Elo Viiding ; aus dem Estnischen von Gisbert Jänicke

    Index Scriptorium Estoniae

    Viiding, Elo, 1974-

    2002-01-01

    Sisu : Die Möglichkeit des Meteorologen = Meteoroloogi võimalikkusest ; "Der Meteorologe kam 1990 in die Stadt..." = "Meteoroloog saabus linna aastal 1990..." ; "Was wäre dir "Arbeit" des Meteorologen..." = "Mis oleks meteoroloogi töö..." ; "Und ein Unglück für den Meteorologen ist es auch..." = "Ja Meteoroloogi õnnetus on veel see..." ; Angst vor dem Altwerden des Meteorologen = Hirm Meteoroloogi vanakssaamise ees ; Fest. Geschenk = Pidu. Kink ; "Wenn der Meteorologe eine Grösse sieht, ist er darüber..." = "Kui meteoroloog näeb suurust, on ta selle kohal..." ; Der Meteorologe wird im Saal erwartet = Meteoroloogi oodatakse saali ; "Das Abkommen mit der Meteorologenerwartung kündigen..." = "Katkestada leping meteoroloogiootusega..." ; "Die "Wege des Herrn" sind der Meteorologe..." = "Looja tee" on Meteoroloog..." ; Von dem Fremden, der im Saal den Meteorologen traf = Võõra lugu, kes Meteoroloogi saalis kohtas ; "Den Fremden hervorzuhusten, der von dem..." = "Köhida enesest välja võõras, kes tahtis teha..." ; Der Fremde beruhigt sich nicht = Võõras ei jää rahule

  3. Uroboros : [aus "Uroboros"] : [luuletused] / Ene Mihkelson ; tlk. Irja Grönholm ja Gisbert Jänicke

    Index Scriptorium Estoniae

    Mihkelson, Ene, 1944-2017-

    2006-01-01

    Sisu: Uroboros : "In der Nacht fragt heimlich jemand Wie heisst du..." ; "Am Meer stand ein Mann mit einer Sense..." ; "An dem Tag als es Kugeln regnete..." ; "Zum Zeitvertrieb ersteigerte ich mir auf einer Auktion ein paar..." ; "Sie hat sich mit der Zeit eingelassen und daraus..." ; Das Feld ; "Mutter ist wieder bei mir obgleich..." ; "Wir waren ein Mädchen Wir redeten..." ; "Als ich vier Jahre alt wurde Da wurde so viel..." ; "Wer ist dein Vater Der Fremde aus der Ferne..." ; "Das letzte Wort Das letzte Das letzte Das allein...". Orig.: Uroboros : "Öö aegu keegi küsib sala Mis su nimi on..." ; "Mere ees seisis vikatiga mees..." ; "Sel päeval kui sadas kuulivihma..." ; "Ajaviiteks ostsin enampakkumisel sõpru..." ; "Ta ajaga läinud on sohki ja sellest..." ; Põld ; "Mu ema on jälle mu juures kuigi..." ; "Meid oli üks türduk Me rääkisime..." ; "Kui olin saanud neljaseks Nii väga kiitis..." ; "Kes on su isa Kaugelt tulnud võõras..." ; "Viimast sõna Viimast Viimast Ainu..."

  4. comparative analysis of calorific value of briquettes produced

    African Journals Online (AJOL)

    yanky

    4.30% at 40% starch level while Daniella oliveri + Rice husk briquettes at 20% starch level had the ... transportation, handling and storage (Husan et al. 2002). .... Table 1: Combustion Properties of Briquettes at 20% Starch Level. Biomass.

  5. Évaluation de l'activité antimicrobienne de quatre feuilles utilisées ...

    African Journals Online (AJOL)

    SARAH

    30 nov. 2015 ... Objectives: Plant leaves used as food packaging having thus a ... Daniellia oliveri and Thalia geniculata used as food packaging preserve better foods during storage. Extract ...... of selected essential oil components on Gram.

  6. Nickel(I) and nickel(III) complexes of substituted tetraaza macrocycles formed by pulse radiolysis and electrochemistry of nickel(II) precursors

    International Nuclear Information System (INIS)

    Bernhardt, P.V.; Lawrance, G.A.; Sangster, D.F.

    1988-01-01

    The square-planar nickel(II) complexes of the ligands 8-methyl-8-nitro-1,3,6,10,13,15-hexaazatricyclo[13.1.1.1/sup 13,15/]octadecane, 8-amino-8-methyl-1,3,6,10,13,15-hexaazatricyclo[13.1.1.1/sup 13,15/]octadecane, 3,7-bis(2-aminoethyl)-1,3,5,7-tetraazabicyclo[3.3.1]nonane, and 9-methyl-9-nitro-1,4,7,11-tetraazacyclotridecane (I-IV) react rapidly with hydroxyl radicals and aquated electrons (e/sub aq/). The initial transient products of these reactions decay via first-order kinetics within a few milliseconds in neutral aqueous solution at 22/degrees/C in all cases. Electronic spectra and decay rate constants, as well as formation rate constants, are reported for all transients. Reaction of the nitro-substituted complexes with e/sub aq/ led to electron addition to the nitro group rather than to the metal center; otherwise, a Ni/sup I/ transient is observed. Following reaction with OH, the product of the initial decay remains a Ni/sup III/ species. This is more long-lived, and stabilization of Ni/sup III/ by axial coordination of the pendant amine in II is indicated. No notable stabilization of Ni/sup I/ or Ni/sup III/ from the presence of the bicyclic azamethylene football in I-III occurs. Cyclic voltammetry in acetonitrile identified both one-electron oxidation and one-electron reduction processes for the nickel(II) complexes, as well as nitro group reduction, where this group was pendant to the macrocycle. 34 references, 3 figures, 3 tables

  7. Assaying Break and Nick-Induced Homologous Recombination in Mammalian Cells Using the DR-GFP Reporter and Cas9 Nucleases

    NARCIS (Netherlands)

    Vriend, Lianne E. M.; Jasin, Maria; Krawczyk, Przemek M.

    2014-01-01

    Thousands of DNA breaks occur daily in mammalian cells, including potentially tumorigenic double-strand breaks (DSBs) and less dangerous but vastly more abundant single-strand breaks (SSBs). The majority of SSBs are quickly repaired, but some can be converted to DSBs, posing a threat to the

  8. Die Form der Zeit : [aus "Aja kuju"¡ ("Die Form der Zeit")] : [luuletused] / Doris Kareva ; tlk. Irja Grönholm ja Gisbert Jänicke

    Index Scriptorium Estoniae

    Kareva, Doris, 1958-

    2006-01-01

    Sisu: Die Form der Zeit : "Die lodernde, reglose Sonne..." ; "Was ist, kann ausgedrückt werden..." ; "Ich höre Stunde um Stunde..." ; "Ich ging am Strand entlang..." ; "Um sichtbarer zu leben..." Der beste Teil des Tags ist die Nacht..." ; "Nächte verschreiben sich dem Morgen..." ; "Herb, karg ist das Licht des Nordens..." ; "Alles rotiert, alles pulsiert..." ; "Sprache ist in der Tat das Haus des Seins..." ; "Der Schmetterling, dessen Flügelschlag..." ; "Die blendende, peinlich reine, bodenlose...". Orig.: Aja kuju : "Leekiv liikumatu päike..." ; "See, mis on, on väljendatav..." ; "Kuulan tunde ja tunde..." ; "Kõndisin mere ääres..." ; "Selleks, et elada selgemalt..." ; "Päeva parim osa on öö..." ; "Ööd kirjutavad end hommikuks..." ; "Karm, napp on põhjamaa valgus..." ; "Kõik tiirleb, kõik tuksleb..." ; "Keel on tõesti olemise koda..." ; "Liblikas, kelle tiivalöök..." ; "Lume pimestav, piinavalt puhas..."

  9. Srovnání organizačních struktur vybraných mládežnických klubů

    OpenAIRE

    Vavříček, Jaroslav

    2006-01-01

    and keywords Title: Comparison of organizational structures of selected youth clubs The Goals: Specify the terms connected with the thesis, work out the methodology part, introduce selected clubs and their organizational structures, interview members and employees ofthese dubs, present my idea. Methodology: Organizational structures of selected soccer dubs and the ways oftheir funding. Suggestion of an idea! organizational structure, informations also acquirred by nonstandard interviews with ...

  10. Lider fallen zu wie die Tore der Burg : [luuletused kogust "Simunapäev"] / Kristiina Ehin ; aus dem Estnischen von Gisbert Jänicke

    Index Scriptorium Estoniae

    Ehin, Kristiina, 1977-

    2004-01-01

    Sisu: die nachricht von der sanktgeorgsnacht = Jüriöö sõnum ; sie standen unter einem schiefen dach = nad seisid viltuse katuse all ; "der soldat steht auf dem hof ..." = "püssimees seisab õues..." ; "der mond heult..." = "kuu ulub..." ; der frauenräuber = naisteröövija ; "ich schlafe als schlamm im wald..." = "magan mudana metsas..." ; "ruhig erhebt sich der weihrauchdunst..." = "vaikselt tõuseb taevasse..."

  11. An energy source which has come just in the nick of time; Une source d'energie qui tombe pile

    Energy Technology Data Exchange (ETDEWEB)

    Jacquemin, Ch.

    1999-12-01

    Fifty-six projects from industrialists and public research organisms have been proposed to the 'fuel cell' research and technological innovation network. Moreover, a project carried out by the French PSA/Renault car manufacturers and devoted to the development of vehicles equipped with fuel cells has been labelled by the network. This short paper summarizes the aim and organization of the fuel cell network in the French scientific and technical program. (J.S.)

  12. Der Handgänger : [aus "Kaitseala" ("Schutzgebiet")] : [luuletused] / Kristiina Ehin ; tlk. Irja Grönholm ja Gisbert Jänicke

    Index Scriptorium Estoniae

    Ehin, Kristiina, 1977-

    2006-01-01

    Luuletusi tsüklist "Kätelkäija", kogumikust Kaitseala : Mohni 2004-2005 (Tallinn : Huma, 2005). Sisu: Der Handgänger ; "draussen ist üppiger juli..." ; "ich menschliche hülle keuche über die erde..." ; "die einsamen waldseen..." ; "wieso finden wir zueinander..." ; "die erste rätsel betrifft die wahrnehmung..." ; "die uhr schlägt für dich ind für mich die stunde des abschieds..." ; weisen, die mein herz dahintrugen 1.-2. Orig.: Kätelkäija ; "väljas on lopsakas juuli..." ; "ihulise inimesena hingeldan mööda maad..." ; "üksildased metsajärved..." ; "miks jõuame teineteiseni..." ; "esimene mõistatus puudutab taju..." ; "kell lööb minu ja sinu siit lahkumise tundi..." ; viisid mis viisid mu südame 1.-2.

  13. Nachtvögel. Nachtgedanken : Gedichte aus den Jahren 1995-1997 / Jaan Kaplinski ; tõlk. Gisbert Jänicke

    Index Scriptorium Estoniae

    Kaplinski, Jaan, 1941-

    1999-01-01

    Luuletused J. Kaplinski kogust "Öölinnud. Öömõtted". Tekst saksa ja eesti k. Sisu: "Über alles ist schon geschrieben, über alles schon gesungen..." = "Kõigest on kirjutatud, kõigest on lauldud..." ; "Die Blumen, durch die ich gesprochen..." = "Need lilled, mille läbi ma olen rääkinud..." ; "Du sagst, das Leichte besiegt am Ende immer das Schwere..." = "Sa ütlesid, et kergus võidab lõpuks ikka raskuse..." ; "In allem, was du jemals..." = "Kõiges, mis sa kunagi oled kirjutanud..." ; "Mondschöpfungsnacht. Ich wasche meine füsse im Dunkeln auf dem Steg. Warm ist es..." = "Kuuloomise öö. Pesen jalgu pimedas purdel. Soe..." ; "Immer mehr Autos, immer weniger Kühe..." = "Autosid ikka rohkem, lehmi vähem..."

  14. Vorliegende Gedichte stammen aus dem Zyklus "Der Handgänger" : [luuletused] / Kristiina Ehin ; eesti keelest tlk Gisbert Jänicke ja Irja Grönholm

    Index Scriptorium Estoniae

    Ehin, Kristiina, 1977-

    2006-01-01

    Luuletusi tsüklist "Kätelkäija" (Kaitseala. Tallinn : Huma, 2005). - Andmeid autori kohta lk. 61. Sisu: Der Handgänger = Kätelkäija : "draussen ist üppiger juli..." = "väljas on lopsakas juuli..." ; "ich menschliche hülle keuche über die erde..." = "ihulise inimesena hingeldan mööda maad..." ; "die einsamen waldseen..." = "üksildased metsajärved..." ; "im schüttelfrost kam sie ans ufer..." = "kaldalejõudmine külmavärinais..."

  15. Production potential and stability of a broadleaved mixed oak/hornbeam forest stand situated on a eutrophic site, Ždánický les

    Directory of Open Access Journals (Sweden)

    Václav Hurt

    2012-01-01

    Full Text Available The paper focuses on assessing the growth and production of a mixed oak/hornbeam forest stand established by combined regeneration in 1940 to 1942. The stand is situated at an altitude of 460 m. Since 1961, it is left to its natural development. The 25–year–old stand was characterized as an individually mixed, both diameter- and height-differentiated pole-stage stand. The proportion of tree species was as follows: sessile oak 77 %, hornbeam 19 %, birch 1 %, lime 1 %, black poplar 1 %, wild cherry tree, wild service tree, and field maple. During 41 years of measurements, the proportion of oak slightly decreased to 76 %, on the other hand, the proportion of hornbeam increased to 22%. The initial growing stock of the 25–year–old stand, 75 m3.ha−1, increased to 323 m3.ha−1 at an age of 66 years in 2008. At present, current volume increment ranged between 6.3 m3.ha−1.year−1 and 11.6 m3.ha−1.year−1 during years 1967 and 1998. Since the age of 61, the growth of the stand has decreased and then even ceased due to increased mortality of oak.

  16. Nářeční lexikum na východní Moravě a karpatská salašnická kolonizace

    Czech Academy of Sciences Publication Activity Database

    Goláňová, Hana

    -, č. 4 (2016), s. 316-327 ISSN 0862-8351 R&D Projects: GA ČR(CZ) GA16-04648S Institutional support: RVO:68378092 Keywords : Carpathian shepherd colonization * Obščekarpatskij dialektologičeskij atlas * dialectal lexicon * carpathism * eastern-Moravian dialect Subject RIV: AI - Linguistics

  17. Scope and Limitations of the Nicking Enzyme Amplification Reaction for the Synthesis of Base-Modified Oligonucleotides and Primers for PCR

    Czech Academy of Sciences Publication Activity Database

    Ménová, Petra; Raindlová, Veronika; Hocek, Michal

    2013-01-01

    Roč. 24, č. 6 (2013), s. 1081-1093 ISSN 1043-1802 R&D Projects: GA ČR GA203/09/0317 Institutional support: RVO:61388963 Keywords : isothermal DNA amplification * cross - coupling reactions * nucleoside triphosphates * polymerase incorporation * functionalized DNA * nucleic-acids Subject RIV: CC - Organic Chemistry Impact factor: 4.821, year: 2013

  18. Stille wird zu Farben : [aus "Vaikus saab värvideks" ("Stille wird zu Farben")] : [luuletused] / Jaan Kaplinski ; tlk. Irja Grönholm ja Gisbert Jänicke

    Index Scriptorium Estoniae

    Kaplinski, Jaan, 1941-

    2006-01-01

    Sisu: Stille wird zu Farben ; Der letzte rote Apfel ; Wer allein bleibt ; Ruf nach dem Vorherigen ; Mit jedem Frühling ; So viele Namen ; In die Spuren ; Höher ; Es wird Licht ; Mit dem Sommerwrind um die Wette ; Ob sie noch hört. Orig.: Vaikus saab värvideks ; Viimane verev õun ; Üksi jääb ; Hüüab eelmist ; Iga kevadega ; Niipalju nimesid ; Astub tuule jälgedele ; Käib kõrgemalt ; Saab valguseks ; Suvetuulega võidu ; Kas kuuleb enam

  19. A direct view by immunofluorescent comet assay (IFCA) of DNA damage induced by nicking and cutting enzymes, ionizing (137)Cs radiation, UV-A laser microbeam irradiation and the radiomimetic drug bleomycin.

    Science.gov (United States)

    Grigaravicius, Paulius; Rapp, Alexander; Greulich, Karl Otto

    2009-03-01

    In DNA repair research, DNA damage is induced by different agents, depending on the technical facilities of the investigating researchers. A quantitative comparison of different investigations is therefore often difficult. By using a modified variant of the neutral comet assay, where the histone H1 is detected by immunofluorescence [immunofluorescent comet assay (IFCA)], we achieve previously unprecedented resolution in the detection of fragmented chromatin and show that trillions of ultraviolet A photons (of a few eV), billions of bleomycin (BLM) molecules and thousands of gamma quanta (of 662 keV) generate, in first order, similar damage in the chromatin of HeLa cells. A somewhat more detailed inspection shows that the damage caused by 20 Gy ionizing radiation and by a single laser pulse of 10 microJ are comparable, while the damage caused by 12 microg/ml BLM depends highly on the individual cell. Taken together, this work provides a detailed view of DNA fragmentation induced by different treatments and allows comparing them to some extent, especially with respect to the neutral comet assay.

  20. Der allerschönste Krieg : [aus "Kõige ilusam sõda"¡ ("Der allerschönste Krieg")] : [luuletused] / Asko Künnap ; tlk. Irja Grönholm ja Gisbert Jänicke

    Index Scriptorium Estoniae

    Künnap, Asko

    2006-01-01

    Sisu: Der allerschönste Krieg : Ich suchte mein nichtgelebtes Leben ; Wie fühlt sich's Seite an Seite ; So toll, dass wir kommen konnten ; Wie kommt man lebend aus Schweden zurück? ; Im Schlaf vereint ; Ruft die Spinnen - dass sie die Nacht weben ; Du hältst die Welt von mir fern ; Ruder mich zur Insel ; In den Augen des Mädchens aus dem Norden ; Der allerschönste Krieg ; Der Teufel im Sakko zu sechshundert Kronen. Orig.: Kõige ilusam sõda : Otsisin oma elamata jäänud elu ; Kuidas on kõrvuti olla ; Nii tore, et tulla saime ; Kuidas tulla elusana tagasi Rootsist? ; Unes ühes ; Kutsuge ämblikud - las punuvad öö ; Sina hoiad maailma minust ära ; Sõua mind saareni ; Ühe põhjamaa tüdruku silmades ; Kõige ilusam sõda ; Kurat kuuesajakroonises pintsakus

  1. Adresát Jindřich Honzl. Dopisy a scénické návrhy Jindřicha Štyrského z Honzlovy pozůstalosti

    Czech Academy of Sciences Publication Activity Database

    Bydžovská, Lenka; Srp, K.

    2008-01-01

    Roč. 56, č. 6 (2008), s. 529-537 ISSN 0049-5123 R&D Projects: GA AV ČR IAA800330603 Institutional research plan: CEZ:AV0Z80330511 Keywords : stage design * artificialism * surrealism Subject RIV: AL - Art, Architecture, Cultural Heritage

  2. Effect of feeding some evergreen tropical browse plant leaves on ...

    African Journals Online (AJOL)

    A feeding trial was conducted with thirty (30) weaner rabbits to investigate the nutritive potentials of some evergreen tropical browse plant leaves (Ficcus thoningii, Vitex doniana, Daniela oliveri, Sarcocephalus latifolia). Mixed breed rabbits were used and randomly assigned to five (5) treatments (T1 - T5). The rabbits in ...

  3. Author Details

    African Journals Online (AJOL)

    Tope, SF. Vol 4, No 4 (2010) - Articles Foraging behaviour of Apis mellifera adansonii Latreille (Hymenoptera : Apidae) on Daniellia oliveri, Delonix regia, Hymenocardia acida and Terminalia mantaly flowers in Ngaoundéré (Cameroon) Abstract PDF. ISSN: 1997-342X. AJOL African Journals Online. HOW TO USE AJOL.

  4. Author Details

    African Journals Online (AJOL)

    Mbianda, AP. Vol 4, No 4 (2010) - Articles Foraging behaviour of Apis mellifera adansonii Latreille (Hymenoptera : Apidae) on Daniellia oliveri, Delonix regia, Hymenocardia acida and Terminalia mantaly flowers in Ngaoundéré (Cameroon) Abstract PDF. ISSN: 1997-342X. AJOL African Journals Online. HOW TO USE AJOL ...

  5. 1809-IJBCS-Article-Hoekou Yao

    African Journals Online (AJOL)

    hp

    Original Paper http://indexmedicus.afro.who.int. Évaluation des propriétés antimicrobiennes de Pterocarpus erinaceus Poir. (Faboïdeae) et Daniellia oliveri (Rolfe) Hutch. et Dalz (Caesalpinoïdeae), utilisées en médecine traditionnelle au Togo. Nassifatou Koko TITTIKPINA 1, Amégninou AGBAN 2,Koffi Apeti GBOGBO 1,.

  6. In vitro anthelmintic effect of two medicinal plants (Anogeissus ...

    African Journals Online (AJOL)

    STORAGESEVER

    2009-09-15

    Sep 15, 2009 ... A study was conducted to evaluate Anogeissus leiocarpus leaf and Daniellia oliveri stem barks as effective remedy for gastrointestinal parasites. The anthelmintic activity of these extracts on eggs, first stage larvae and adults of Haemonchus contortus was examined by in vitro tests. The extracts were.

  7. Further Thoughts on "How Task Features Impact Evidence from Assessments Embedded in Simulations and Games"

    Science.gov (United States)

    Oliveri, María Elena; Khan, Saad

    2014-01-01

    María Oliveri, and Saad Khan write that the article: "How Task Features Impact Evidence from Assessments Embedded in Simulations and Games" provided helpful illustrations regarding the implementation of evidence-centered assessment design (Mislevy & Haertel, 2006; Mislevy, Steinberg, & Almond, 1999) with games and simulations.…

  8. [Characterization of the genetic variability of field strains of Brucella canis isolated in Antioquia].

    Science.gov (United States)

    Vidal Arboleda, Juana L; Ortiz Roman, Luisa F; Olivera Angel, Martha

    2017-12-22

    Brucella canis is a facultative intracellular pathogen responsible for canine brucellosis, a zoonotic disease that affects canines, causing abortions and reproductive failure; and the production of non-specific symptoms in humans. In 2005 the presence of B. canis in Antioquia was demonstrated and the strains were identified as type 2. The sequencing of the genome of a field strain denoted Brucella canis str. Oliveri, showed species-specific indel events, which led us to investigate the genomic characteristics of the B. canis strain isolated and to establish the phylogenetic relationships and the divergence time of B. canis str. Oliveri. Conventional PCR sequencing was performed in 30 field strains identifying 5 indel events recognized in B. canis str. Oliveri. ADN from Brucella suis, Brucella melitensis and vaccine strains from Brucella abortus were used as control, and it was determined that all of the studied field strains shared 4 out of the 5 indels of the sequenced Oliveri strain, indicating the presence of more than one strain circulating in the region. Phylogenetic analysis was performed with 24 strains of Brucella using concatenated sequences of genetic markers for species differentiation. The molecular clock hypothesis and Tajima's relative rate test were tested, showing that the Oliveri strain, similarly to other canis species, diverged from B. suis. The molecular clock hypothesis between Brucella species was rejected and an evolution rate and a similar genetic distance between the B. canis were demonstrated. Copyright © 2017 Asociación Argentina de Microbiología. Publicado por Elsevier España, S.L.U. All rights reserved.

  9. Low Ultraviolet B and Increased Risk of Brain Cancer: An Ecological Study of 175 Countries

    Science.gov (United States)

    2009-07-01

    Mean Author Year Journal Location 25(OH)D ng/ml Mazess et al. 1985 Am J Clin Nutr Alaska, USA 16.6 Oliveri et al. 1990 Medicina Buenos Aires...Clin Nutr USA 32.9 Rucker et al. 2002 Cmaj Calgary, Canada 23.8 Tangpricha et al. 2002 Am J Med USA 29.1 Fassi et al. 2003 Medicina Buenos Aires

  10. Cytotoxic geranylated xanthones and O-alkylated derivatives of alpha-mangostin

    DEFF Research Database (Denmark)

    Ha, Ly Dieu; Hansen, Poul Erik; Vang, Ole

    2009-01-01

    Two new geranylated xanthones, 6-O-methylcowanin (4) and oliverixanthone (5), along with five known compounds, cowanin, rubraxanthone, cowaxanthone, cowanol, and β-mangostin, have been isolated from the bark of Garcinia oliveri. For comparison of their biological activities, one mono- and seven di-O-alkylated...... that α-mangostin had the strongest activity, and all the O-alkylated α-mangostin derivatives showed reduced activity compared to the naturally occurring α-mangostin....

  11. Lidový hudebně-taneční projev jako scénický tvar - folklorní soubory v českých zemích v péči jedné instituce

    Czech Academy of Sciences Publication Activity Database

    Stavělová, Daniela

    2015-01-01

    Roč. 25, č. 4 (2015), s. 292-306 ISSN 0862-8351 Institutional support: RVO:68378076 Keywords : folk dance * folk music * stage * show * institution * folklore ensemble * folklore stylization Subject RIV: AC - Archeology, Anthropology, Ethnology

  12. Functional Dissection of the DNA Interface of the Nucleotidyltransferase Domain of Chlorella Virus DNA Ligase*

    Science.gov (United States)

    Samai, Poulami; Shuman, Stewart

    2011-01-01

    Chlorella virus DNA ligase (ChVLig) has pluripotent biological activity and an intrinsic nick-sensing function. ChVLig consists of three structural modules that envelop nicked DNA as a C-shaped protein clamp: a nucleotidyltransferase (NTase) domain and an OB domain (these two are common to all DNA ligases) as well as a distinctive β-hairpin latch module. The NTase domain, which performs the chemical steps of ligation, binds the major groove flanking the nick and the minor groove on the 3′-OH side of the nick. Here we performed a structure-guided mutational analysis of the NTase domain, surveying the effects of 35 mutations in 19 residues on ChVLig activity in vivo and in vitro, including biochemical tests of the composite nick sealing reaction and of the three component steps of the ligation pathway (ligase adenylylation, DNA adenylylation, and phosphodiester synthesis). The results highlight (i) key contacts by Thr-84 and Lys-173 to the template DNA strand phosphates at the outer margins of the DNA ligase footprint; (ii) essential contacts of Ser-41, Arg-42, Met-83, and Phe-75 with the 3′-OH strand at the nick; (iii) Arg-176 phosphate contacts at the nick and with ATP during ligase adenylylation; (iv) the role of Phe-44 in forming the protein clamp around the nicked DNA substrate; and (v) the importance of adenine-binding residue Phe-98 in all three steps of ligation. Kinetic analysis of single-turnover nick sealing by ChVLig-AMP underscored the importance of Phe-75-mediated distortion of the nick 3′-OH nucleoside in the catalysis of DNA 5′-adenylylation (step 2) and phosphodiester synthesis (step 3). Induced fit of the nicked DNA into a distorted conformation when bound within the ligase clamp may account for the nick-sensing capacity of ChVLig. PMID:21335605

  13. Functional dissection of the DNA interface of the nucleotidyltransferase domain of chlorella virus DNA ligase.

    Science.gov (United States)

    Samai, Poulami; Shuman, Stewart

    2011-04-15

    Chlorella virus DNA ligase (ChVLig) has pluripotent biological activity and an intrinsic nick-sensing function. ChVLig consists of three structural modules that envelop nicked DNA as a C-shaped protein clamp: a nucleotidyltransferase (NTase) domain and an OB domain (these two are common to all DNA ligases) as well as a distinctive β-hairpin latch module. The NTase domain, which performs the chemical steps of ligation, binds the major groove flanking the nick and the minor groove on the 3'-OH side of the nick. Here we performed a structure-guided mutational analysis of the NTase domain, surveying the effects of 35 mutations in 19 residues on ChVLig activity in vivo and in vitro, including biochemical tests of the composite nick sealing reaction and of the three component steps of the ligation pathway (ligase adenylylation, DNA adenylylation, and phosphodiester synthesis). The results highlight (i) key contacts by Thr-84 and Lys-173 to the template DNA strand phosphates at the outer margins of the DNA ligase footprint; (ii) essential contacts of Ser-41, Arg-42, Met-83, and Phe-75 with the 3'-OH strand at the nick; (iii) Arg-176 phosphate contacts at the nick and with ATP during ligase adenylylation; (iv) the role of Phe-44 in forming the protein clamp around the nicked DNA substrate; and (v) the importance of adenine-binding residue Phe-98 in all three steps of ligation. Kinetic analysis of single-turnover nick sealing by ChVLig-AMP underscored the importance of Phe-75-mediated distortion of the nick 3'-OH nucleoside in the catalysis of DNA 5'-adenylylation (step 2) and phosphodiester synthesis (step 3). Induced fit of the nicked DNA into a distorted conformation when bound within the ligase clamp may account for the nick-sensing capacity of ChVLig.

  14. "Nepřipustíme, aby nedostavěný most zdržel přijíždějící vlak!". Postoj mládeže k přírodě během budování mládežnických železničních tratí v Československu a Jugoslávii (1946-1949)

    Czech Academy of Sciences Publication Activity Database

    Sovilj, Milan

    2016-01-01

    Roč. 38, č. 4 (2016), s. 36-39 ISSN 0418-5129 R&D Projects: GA ČR GA15-04902S Institutional support: RVO:68378114 Keywords : Czechoslovakia * Yugoslavia * youth railwyas * history of nature Subject RIV: AB - History

  15. Characterization of linear forms of the circular enterocin AS-48 obtained by limited proteolysis

    NARCIS (Netherlands)

    Montalbán-López, Manuel; Spolaore, Barbara; Pinato, Odra; Martínez-Bueno, Manuel; Valdivia, Eva; Maqueda, Mercedes; Fontana, Angelo

    2008-01-01

    AS-48 is a 70-residue circular peptide from Enterococcus faecalis with a broad antibacterial activity. Here, we produced by limited proteolysis a protein species carrying a single nicking and fragments of 55 and 38 residues. Nicked AS-48 showed a lower helicity by far-ultraviolet circular dichroism

  16. Sudha Bhattacharya Jawaharlal Nehru University New Delhi

    Indian Academy of Sciences (India)

    TSC

    Bottom strand. 1. 2. 3. G A T C. 5. 1. 3. 4. 2. *. Nicking profile of EN on the 174 bp substrate. Insertion point. Bottom strand labelled 174bp substrate. Transposon insertion point. Endonuclease nicks at three hot spots. The transposon inserts at hot spot # 3 ...

  17. Alteration of Sequence Specificity of the Type IIS Restriction Endonuclease BtsI

    OpenAIRE

    Guan, Shengxi; Blanchard, Aine; Zhang, Penghua; Zhu, Zhenyu

    2010-01-01

    The Type IIS restriction endonuclease BtsI recognizes and digests at GCAGTG(2/0). It comprises two subunits: BtsIA and BtsIB. The BtsIB subunit contains the recognition domain, one catalytic domain for bottom strand nicking and part of the catalytic domain for the top strand nicking. BtsIA has the rest of the catalytic domain that is responsible for the DNA top strand nicking. BtsIA alone has no activity unless it mixes with BtsIB to reconstitute the BtsI activity. During characterization of ...

  18. Civil Engineering Applications of Ground Penetrating Radar Recent Advances @ the ELEDIA Research Center

    Science.gov (United States)

    Salucci, Marco; Tenuti, Lorenza; Nardin, Cristina; Oliveri, Giacomo; Viani, Federico; Rocca, Paolo; Massa, Andrea

    2014-05-01

    -set-based optimization have been developed for the qualitative reconstruction of multiple and disconnected homogeneous scatterers [5]. Finally, the real-time detection and classification of subsurface scatterers has been investigated by means of learning-by-examples (LBE) techniques, such as Support Vector Machines (SVM) [6]. Acknowledgment - This work was partially supported by COST Action TU1208 'Civil Engineering Applications of Ground Penetrating Radar' References [1] M. Salucci, D. Sartori, N. Anselmi, A. Randazzo, G. Oliveri, and A. Massa, 'Imaging Buried Objects within the Second-Order Born Approximation through a Multiresolution Regularized Inexact-Newton Method', 2013 International Symposium on Electromagnetic Theory (EMTS), (Hiroshima, Japan), May 20-24 2013 (invited). [2] A. Giannopoulos, 'Modelling ground penetrating radar by GprMax', Construct. Build. Mater., vol. 19, no. 10, pp.755 -762 2005 [3] L. Poli, G. Oliveri, P. Rocca, and A. Massa, "Bayesian compressive sensing approaches for the reconstruction of two-dimensional sparse scatterers under TE illumination," IEEE Trans. Geosci. Remote Sensing, vol. 51, no. 5, pp. 2920-2936, May. 2013. [4] L. Poli, G. Oliveri, and A. Massa, "Imaging sparse metallic cylinders through a Local Shape Function Bayesian Compressive Sensing approach," Journal of Optical Society of America A, vol. 30, no. 6, pp. 1261-1272, 2013. [5] M. Benedetti, D. Lesselier, M. Lambert, and A. Massa, "Multiple shapes reconstruction by means of multi-region level sets," IEEE Trans. Geosci. Remote Sensing, vol. 48, no. 5, pp. 2330-2342, May 2010. [6] L. Lizzi, F. Viani, P. Rocca, G. Oliveri, M. Benedetti and A. Massa, "Three-dimensional real-time localization of subsurface objects - From theory to experimental validation," 2009 IEEE International Geoscience and Remote Sensing Symposium, vol. 2, pp. II-121-II-124, 12-17 July 2009.

  19. 75 FR 71787 - Advisory Committee On Historical Diplomatic Documentation; Notice of Meeting

    Science.gov (United States)

    2010-11-24

    ... session must notify Nick Sheldon, Office of the Historian (202-663-1123) no later than December 9, 2010... of State, Office of the Historian, Washington, DC 20520, telephone (202) 663-1123, (e-mail history...

  20. 76 FR 53165 - Advisory Committee on Historical Diplomatic Documentation; Notice of Meeting

    Science.gov (United States)

    2011-08-25

    ... planning to attend must notify Nick Sheldon, Office of the Historian (202-663-1123) no later than September... of State, Office of the Historian, Washington, DC 20520, telephone (202) 663-1123, (e-mail history...

  1. 76 FR 9071 - Advisory Committee on Historical Diplomatic Documentation; Notice of Meeting

    Science.gov (United States)

    2011-02-16

    ... notify Nick Sheldon, Office of the Historian (202-663-1123) no later than March 3, 2011 to provide date..., Advisory Committee on Historical Diplomatic Documentation, Department of State, Office of the Historian...

  2. 76 FR 28844 - Advisory Committee on Historical Diplomatic Documentation; Notice of Meeting

    Science.gov (United States)

    2011-05-18

    ... notify Nick Sheldon, Office of the Historian (202-663-1123) no later than June 2, 2011 to provide date of..., Department of State, Office of the Historian, Washington, DC 20520, telephone (202) 663-1123, (e-mail history...

  3. Will we ever see their like again? Departing directors at the top US physics laboratories leave a discipline in search of inspired leadership

    CERN Multimedia

    1998-01-01

    Burt Richter stands down next August as director of the Stanford Linear Accelerator Centre. His departure follows that of Nick Samios from Brookhaven National Laboratory and the retirement next year of John Peoples from Fermilab.

  4. 78 FR 67927 - Foreign Trade Regulations (FTR): Mandatory Automated Export System Filing for All Shipments...

    Science.gov (United States)

    2013-11-13

    ... Internet at [email protected] ). FOR FURTHER INFORMATION CONTACT: Nick Orsini, Chief, Foreign Trade Division... 0607-AA50 Foreign Trade Regulations (FTR): Mandatory Automated Export System Filing for All Shipments... approval of new information collection requirements. [[Page 67928

  5. Plaadid / Rein Vader, Heidi Purga, Tiiu Laks...[jt.

    Index Scriptorium Estoniae

    2008-01-01

    Uutest heliplaatidest Nick Cave & The Bad Seeds "Dig, Lazarus, Dig!", Erykah Badu "New Amerikah Part One", Leslie da Bass "Nights By Open Windows", Jack Johnson "Sleep Througth The Static", Janet Jackson "Discipline", "Rock & Roll City"

  6. Pop / Siim Nestor

    Index Scriptorium Estoniae

    Nestor, Siim, 1974-

    2008-01-01

    Heliplaatidest: Janet Jackson "Discipline", Jack Johnson "Sleep Through The Static", The Mars Volta "The Bedlam in Goliath", Christopher Clark "Turning Dragon", Erykah Badu "New Amerykah Part One (4th World War)", Nick Cave & The Bad Seeds "Dig, Lazarus, Dig!!!"

  7. Folk, akustiline selline, on hambus Tristan Priimäel / Tristan Priimägi

    Index Scriptorium Estoniae

    Priimägi, Tristan, 1976-

    2004-01-01

    Heliplaatidest: Iron & Wine "Our Endless Numbered Days", Sufjan Stevens "Seven Swans", Joanna Newsom "The Milk-Eyed Mender", Nick Drake "Made to Love Magic", Kings Of Convenience "Riot On an Empty Street", White Magic "Through the Sun Door"

  8. Pop / Tauno Maarpuu

    Index Scriptorium Estoniae

    Maarpuu, Tauno

    2006-01-01

    Heliplaatidest: Scandinavian Music Group "Hölmö rakkaus", Baron Zen "At The Mall", Cave, Nick & Warren Ellis "The Proposition. Original Soundtrack", Eläkeläiset "Humppasirkus", Warren Zevon "Reconsider Me: The Love Songs"

  9. Pre-Processing Data Using Wavelet Transform and PCA Based on ...

    Indian Academy of Sciences (India)

    5

    wavelet theory in the last century, using this tool in the hydrology has been an ... These models are based on Statistical Learning Theory, which is ...... Hutcheson G and Nick S 1999 The multivariate social scientist: Introductory statistics using.

  10. A multiverse play divides opinion

    Science.gov (United States)

    Crease, Robert P.

    2015-03-01

    The stage lights rise. A man and woman meet in a cute way - "Do you know why it's impossible to lick the tips of your elbows?" she asks - they chat momentarily, and separate. The play is Constellations by Nick Payne.

  11. Sequence Classification: 390305 [

    Lifescience Database Archive (English)

    Full Text Available SOMERASE I TOPA (OMEGA-PROTEIN) (RELAXING ENZYME) (UNTWISTING ENZYME) (SWIVELASE) (TYPE I DNA TOPOISOMERASE) (NICKING-CLOSING ENZYME) || http://www.ncbi.nlm.nih.gov/protein/31794816 ...

  12. Nicholas J. Nagle | NREL

    Science.gov (United States)

    @nrel.gov | 303-384-6184 Research Interests Nick is interested in understanding how advanced feedstocks Technology (2015) "Acid-catalyzed algal biomass pretreatment for integrated lipid and carbohydrate-based

  13. Pop / Mart Juur

    Index Scriptorium Estoniae

    Juur, Mart, 1964-

    2005-01-01

    Heliplaatidest: Nick Cave & The Bad Seeds "B-Sides & Rarities", Koneveljet "Hoax", Everything But The Girl "Adapt or Die. Ten Years of Remixes", Autechre "Untitled", Daedelus "Exquisite Corpse", Keane "Live recordings 2004", Iris Garrelfs "Specified Encounters"

  14. Vene elektrivõrk : Gazprom on ebakindel gaasitarnija / Raimo Poom

    Index Scriptorium Estoniae

    Poom, Raimo

    2007-01-01

    Tallinnas toimunud rahvusvahelisel energiajulgeoleku konverentsil räägiti energia- ja keskkonnateemadel. Vene elektrivõrkude esindaja Marina Liborakina hoiatas Gazpromi vigade eest gaasi tarnimisel. Kommenteerib Briti nõustamisfirma E3G juhataja Nick Mabey

  15. Site-specifically modified oligodeoxyribonucleotides as templates for Escherichia coli DNA polymerase I

    International Nuclear Information System (INIS)

    O'Connor, D.; Stoehrer, G.

    1985-01-01

    Oligodeoxyribonucleotides with site-specific modifications have been used as substrates for Escherichia coli DNA polymerase I holoenzyme and Klenow fragment. Modifications included the bulky guanine-8-aminofluorene adduct and a guanine oxidation product resembling the product of photosensitized DNA oxidation. By a combination of primers and nick-mers, conditions of single-strand-directed DNA synthesis and nick-translation could be created. The results show that the polymerase can bypass both types of lesions. Bypass occurs on a single-stranded template but is facilitated on a nicked, double-stranded template. Only purines, with guanine more favored than adenine, are incorporated across both lesions. The results indicate that site-specifically modified oligonucleotides can be sensitive probes for the action of polymerases on damaged templates. They also suggest a function for polymerase I, in its nick-translation capacity, during DNA repair and mutagenesis

  16. Book Review: A Liberal Actor in a Realist World the European Union ...

    African Journals Online (AJOL)

    Abstract. Book Title: A Liberal Actor in a Realist World the European Union Regulatory State and the Global Political Economy of Energy. Book Author: Andreas Goldthau & Nick Sitter. Oxford University Press Oxford 2015. ISBN 9780198719595 ...

  17. Briti asepeaminister arvustas Brüsselis võetud positsiooni / Evelyn Kaldoja

    Index Scriptorium Estoniae

    Kaldoja, Evelyn, 1980-

    2011-01-01

    Suurbritannia asepeaminister Nick Clegg kritiseeris peaminister David Cameroni seisukohti Euroopa Ülemkogul. Suurbritannia otsusest välja jääda Euroopa Liidu uuest valitsustevahelisest rahanduskokkuleppest

  18. Plaadid / Villu Koskaru

    Index Scriptorium Estoniae

    Koskaru, Villu, 1976-

    2003-01-01

    Uutest plaatidest Era "The Mass", Dannii Minogue "Neon Nights", Aaliyah "I Care 4 U", Linkin Park " Meteora", "Smash Hits 6", The Mighty Pop "Yellow", John Taylor "Rosslyn", Nick Cave & The Bad Seds "Nocturama"

  19. Harbor porpoise - Passive acoustic monitoring of harbor porpoises in Puget Sound

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Drs. Marla Holt, Brad Hanson, and Candice Emmons of the NWFSC, along with efforts from the NWFSC dive team (coordinated by Nick Tolimieri), are currently conducting...

  20. Uusi plaate

    Index Scriptorium Estoniae

    2003-01-01

    Heliplaatidest: Tom Jones "Greatest hits", Pet Shop Boys "Disco 3", Epoca De Ouro Ensemble "Cafe Brasil 2", Nick Cave and The Bad Seeds "Nocturama", Pistepirkko "The Nature of 22 Pistepirkko", Jimi Tenor "Higher Planes", "Guitars, Saxes & More", "Inspired 2"

  1. Structure-function analysis of the OB and latch domains of chlorella virus DNA ligase.

    Science.gov (United States)

    Samai, Poulami; Shuman, Stewart

    2011-06-24

    Chlorella virus DNA ligase (ChVLig) is a minimized eukaryal ATP-dependent DNA sealing enzyme with an intrinsic nick-sensing function. ChVLig consists of three structural domains, nucleotidyltransferase (NTase), OB-fold, and latch, that envelop the nicked DNA as a C-shaped protein clamp. The OB domain engages the DNA minor groove on the face of the duplex behind the nick, and it makes contacts to amino acids in the NTase domain surrounding the ligase active site. The latch module occupies the DNA major groove flanking the nick. Residues at the tip of the latch contact the NTase domain to close the ligase clamp. Here we performed a structure-guided mutational analysis of the OB and latch domains. Alanine scanning defined seven individual amino acids as essential in vivo (Lys-274, Arg-285, Phe-286, and Val-288 in the OB domain; Asn-214, Phe-215, and Tyr-217 in the latch), after which structure-activity relations were clarified by conservative substitutions. Biochemical tests of the composite nick sealing reaction and of each of the three chemical steps of the ligation pathway highlighted the importance of Arg-285 and Phe-286 in the catalysis of the DNA adenylylation and phosphodiester synthesis reactions. Phe-286 interacts with the nick 5'-phosphate nucleotide and the 3'-OH base pair and distorts the DNA helical conformation at the nick. Arg-285 is a key component of the OB-NTase interface, where it forms a salt bridge to the essential Asp-29 side chain, which is imputed to coordinate divalent metal catalysts during the nick sealing steps.

  2. Structure-Function Analysis of the OB and Latch Domains of Chlorella Virus DNA Ligase*

    Science.gov (United States)

    Samai, Poulami; Shuman, Stewart

    2011-01-01

    Chlorella virus DNA ligase (ChVLig) is a minimized eukaryal ATP-dependent DNA sealing enzyme with an intrinsic nick-sensing function. ChVLig consists of three structural domains, nucleotidyltransferase (NTase), OB-fold, and latch, that envelop the nicked DNA as a C-shaped protein clamp. The OB domain engages the DNA minor groove on the face of the duplex behind the nick, and it makes contacts to amino acids in the NTase domain surrounding the ligase active site. The latch module occupies the DNA major groove flanking the nick. Residues at the tip of the latch contact the NTase domain to close the ligase clamp. Here we performed a structure-guided mutational analysis of the OB and latch domains. Alanine scanning defined seven individual amino acids as essential in vivo (Lys-274, Arg-285, Phe-286, and Val-288 in the OB domain; Asn-214, Phe-215, and Tyr-217 in the latch), after which structure-activity relations were clarified by conservative substitutions. Biochemical tests of the composite nick sealing reaction and of each of the three chemical steps of the ligation pathway highlighted the importance of Arg-285 and Phe-286 in the catalysis of the DNA adenylylation and phosphodiester synthesis reactions. Phe-286 interacts with the nick 5′-phosphate nucleotide and the 3′-OH base pair and distorts the DNA helical conformation at the nick. Arg-285 is a key component of the OB-NTase interface, where it forms a salt bridge to the essential Asp-29 side chain, which is imputed to coordinate divalent metal catalysts during the nick sealing steps. PMID:21527793

  3. Lack of evidence that the XqYq pairing tips at meiosis in the mouse show hypersensitivity to DNAse I.

    Science.gov (United States)

    Separovic, E R; Chandley, A C

    1987-01-01

    In situ nick translation procedures have been applied to meiotic metaphase I divisions of the normal and XY, Sxr mouse. Unlike in man, where the pairing tips of the XY bivalent show a special sensitivity to DNAse I nicking, no such sensitivity can be detected for either of these types of mouse. Hypersensitivity in the D-band equivalent region of the X chromosome does, however, exist, this site being early replicating in somatic cells and housing the X inactivation centre (Xce).

  4. The preparation of radioactivity labelled DNA

    International Nuclear Information System (INIS)

    Ballance, P.; Morgan, J.; McGregor, G.; Durkacz, B.

    1984-01-01

    The nick translation reaction, which uses the endonuclease enzyme to incorporate pieces of DNA into the genetic material of other organisms, is being used more and more in Molecular Biology research for the preparation of pure DNA labelled with 32 P. However results are presented which show that high radiation doses are received by the hands of nick translation workers. A Scheme of Work to reduce these doses is described. (author)

  5. "Who's There?": Shakespeare and the Dragon of Autism

    Science.gov (United States)

    Renino, Christopher

    2009-01-01

    Dan recently turned 13; Nick will soon be 12. Both boys are smart and fun; both are loving, write like angels, like to learn, and want friends; both are autistic and hope to learn to speak. They find it challenging to relate to and function in this world, and they work hard to improve their abilities to do so. Last fall, Nick and Dan became…

  6. Use of Performance Measures to Evaluate, Document Competence and Deterioration of ASSET Surgical Skills

    Science.gov (United States)

    2016-05-01

    meeting with Advisory Board Member, Prof Nick Sevdalis (London UK) in Seattle at the ASE meeting (April 21-25 2015) to address how to validate our...cognitive task analysis and human factors, and Prof. Nick Sevdalis , an experimental psychologist and expert in patient safety in hospital...Individual Procedure Score (IPS) performance metric and to update Prof Sevdalis on project progress.  Presented a poster and 2 podium presentation at

  7. Crossing the street

    DEFF Research Database (Denmark)

    Shepherd, Nick

    2001-01-01

    Nick Shepherd teaches a class on 'race and racism' at the University of Cape Town. Here he shares the journey he undertakes with his students to find a "civil space'' to discuss sensitive and complex issues.......Nick Shepherd teaches a class on 'race and racism' at the University of Cape Town. Here he shares the journey he undertakes with his students to find a "civil space'' to discuss sensitive and complex issues....

  8. A view from the top: US enrichment Corp.'s William H. Timbers, Jr

    International Nuclear Information System (INIS)

    Giltenan, E.

    1993-01-01

    Nick Timbers took over as the first Transition Manager of the US Enrichment Corporation upon its founding last July 1st. Although USEC is not involved in negotiating the HEU deal, the fledgling company will be in charge of actually buying and selling the resulting LEU. Whenever the deal is finally signed. After the politics and haggling are over, it will be up to Nick Timbers to make the deal work on the global uranium market. The view from USEC is resolute. No matter what shape the final HEU deal takes, Nick Timbers promises that USEC will remain a competitive supplier of enrichment services. Timbers pledges that any extra costs associated with the HEU deal will not be passed on to customers. He took time out from his recent busy schedule to share his thoughts on HEU and its aftermath

  9. Permeabilization of ultraviolet-irradiated chinese hamster cells with polyethylene glycol and introduction of ultraviolet endonuclease from Micrococcus luteus

    International Nuclear Information System (INIS)

    Yarosh, D.B.; Setlow, R.B.

    1981-01-01

    Chinese hamster V-79 cells were made permeable by treatment with polyethylene glycol and then incubated with a Micrococcus luteus extract containing ultraviolet-specific endonuclease activity. This treatment introduced nicks in irradiated, but not in unirradiated, deoxyribonucleic acid. The nicks remained open for at least 3 h; there was no loss of endonuclease-sensitive sites, and no excision of dimers as measured by chromatography was detected. In addition, there was no increase in ultraviolet resistance in treated cells. This suggests that the absence of a significant amount of excision repair in rodent cells is due to the lack of both incision and excision capacity

  10. Substrate specificity of Micrococcus luteus uv endonuclease and its overlap with DNA photolyase activity

    International Nuclear Information System (INIS)

    Patrick, M.H.

    1975-01-01

    The action of an endonuclease from Micrococcus luteus that operates on uv damage in DNA overlaps with that of DNA photolyase from yeast: homo- and heterocyclobutane dipyrimidines in DNA are substrates for both enzymes, but pyrimidine adducts or the spore photoproduct in DNA are not. As expected from this overlap, the action of the two enzymes is mutually interfering: single-strand nicks introduced by the endonuclease effectively preclude photoreactivation; conversely, formation of a photolyase-cyclobutane dipyrimidine complex can prevent nicking by the endonuclease

  11. Hatten af for Cohens skønheder og skyggesider

    DEFF Research Database (Denmark)

    Mortensen, Lise Majgaard

    2011-01-01

    Der er højt til loftet i Aarhus Teaters nye teaterkoncert, som behandler selveste Leonard Cohens lyriske univers. Det er svært at lade være med at sammenligne forestillingen her med Nick Cave Teaterkoncerten, der gik sin sejrsgang i 2007 og 2008. Men det skal man ikke. Instruktøren er den samme....... Men hvor ’Nick Cave’ var rå, grim, provokerende og uanstændig, er ’Leonard Cohen’ en skrøbelig gentleman med en skyggeside....

  12. A conserved helicase processivity factor is needed for conjugation and replication of an integrative and conjugative element.

    Directory of Open Access Journals (Sweden)

    Jacob Thomas

    Full Text Available Integrative and conjugative elements (ICEs are agents of horizontal gene transfer and have major roles in evolution and acquisition of new traits, including antibiotic resistances. ICEs are found integrated in a host chromosome and can excise and transfer to recipient bacteria via conjugation. Conjugation involves nicking of the ICE origin of transfer (oriT by the ICE-encoded relaxase and transfer of the nicked single strand of ICE DNA. For ICEBs1 of Bacillus subtilis, nicking of oriT by the ICEBs1 relaxase NicK also initiates rolling circle replication. This autonomous replication of ICEBs1 is critical for stability of the excised element in growing cells. We found a conserved and previously uncharacterized ICE gene that is required for conjugation and replication of ICEBs1. Our results indicate that this gene, helP (formerly ydcP, encodes a helicase processivity factor that enables the host-encoded helicase PcrA to unwind the double-stranded ICEBs1 DNA. HelP was required for both conjugation and replication of ICEBs1, and HelP and NicK were the only ICEBs1 proteins needed for replication from ICEBs1 oriT. Using chromatin immunoprecipitation, we measured association of HelP, NicK, PcrA, and the host-encoded single-strand DNA binding protein Ssb with ICEBs1. We found that NicK was required for association of HelP and PcrA with ICEBs1 DNA. HelP was required for association of PcrA and Ssb with ICEBs1 regions distal, but not proximal, to oriT, indicating that PcrA needs HelP to progress beyond nicked oriT and unwind ICEBs1. In vitro, HelP directly stimulated the helicase activity of the PcrA homologue UvrD. Our findings demonstrate that HelP is a helicase processivity factor needed for efficient unwinding of ICEBs1 for conjugation and replication. Homologues of HelP and PcrA-type helicases are encoded on many known and putative ICEs. We propose that these factors are essential for ICE conjugation, replication, and genetic stability.

  13. ‘Capiture’ plants with interesting biological activities: a case to go

    Directory of Open Access Journals (Sweden)

    Tittikpina Nassifatou Koko

    2017-10-01

    Full Text Available The investigation of natural products used in Traditional Medicine in Africa is complicated as modern analytical and screening methods are often not available. Computer aided product identification from traditional usage records (CAPITURE may provide an interesting alternative and has been evaluated in the context of an ethnobotanical survey on fungal diseases and their traditional treatment in Tchamba District (Togo. 53 traditional healers were interviewed and their knowledge recorded. Several indicators, the Use Value (UV, Plant Part Value (PPV, Specific Use (SU Value, Intraspecific Use Value (IUV and Informant Consensus Factor (ICF, were applied to the data. Those indices, in addition to a bibliographic review, were then fed into a computer-aided approach which predicted two interesting plants out of the 43 species survey-recorded and their specific activities: Pterocarpus erinaceus sap against ringworm, Daniellia oliveri sap against intertrigo and respectively their roots and trunk barks against candidiasis. Subsequent laboratory-based studies have confirmed the predicted antimicrobial activities with MIC (128 μg/mL to 30 mg/mL and without any notable toxicity on a normal human cell (MRC-5 cells. Although such a method may not be flawless, it is able to provide first leads, and in the face of limited resources, is an attractive alternative worth considering.

  14. Comparison of Auditory Brainstem Response in Noise Induced Tinnitus and Non-Tinnitus Control Subjects

    Directory of Open Access Journals (Sweden)

    Ghassem Mohammadkhani

    2009-12-01

    Full Text Available Background and Aim: Tinnitus is an unpleasant sound which can cause some behavioral disorders. According to evidence the origin of tinnitus is not only in peripheral but also in central auditory system. So evaluation of central auditory system function is necessary. In this study Auditory brainstem responses (ABR were compared in noise induced tinnitus and non-tinnitus control subjects.Materials and Methods: This cross-sectional, descriptive and analytic study is conducted in 60 cases in two groups including of 30 noise induced tinnitus and 30 non-tinnitus control subjects. ABRs were recorded ipsilateraly and contralateraly and their latencies and amplitudes were analyzed.Results: Mean interpeak latencies of III-V (p= 0.022, I-V (p=0.033 in ipsilatral electrode array and mean absolute latencies of IV (p=0.015 and V (p=0.048 in contralatral electrode array were significantly increased in noise induced tinnitus group relative to control group. Conclusion: It can be concluded from that there are some decrease in neural transmission time in brainstem and there are some sign of involvement of medial nuclei in olivery complex in addition to lateral lemniscus.

  15. Pop & rock. Retro. Klassika / Margus Kiis

    Index Scriptorium Estoniae

    Kiis, Margus

    2001-01-01

    Orbital "The Altogether", Vonda Shepard "Ally McBeal (For Once In My Life)", Jääboiler "Niiongi", Tanel Padar ja Speed Free "Woman Knows", Arvo Pärt "Johannes-Passion", Nikka Costa "Everybody Got Their Something", Stevie Nicks "Trouble In Shangri-La", Richard Thompson "Action Packed The Best of the Capitol Years", "Eurovision Song Contest 2001" heliplaatide tutvustused

  16. Technology

    Science.gov (United States)

    Breakstone, Elizabeth

    2008-01-01

    This column presents an interview with Nick Baker, Reference and Web Services librarian at Williams College. Baker has produced several films for and about the library profession. In 2007, he won an InfoTubey Award for Outstanding Library YouTube Productions for his work. In this interview, Baker shares his ideas and experiences about this…

  17. The Significance of Learning Nicknames of Public Figures in Modern English and American Language Models of the World

    Science.gov (United States)

    Garayeva, Almira K.; Akhmetzyanov, Ildar G.; Khismatullina, Lutsia G.

    2016-01-01

    The importance of the topic of this study is determined by several factors: increased interest of linguists to the problem of interaction between language and culture; the need to study the onomastic units as body language. The purpose of this article is to identify the types of motivational nick names of famous American and English public…

  18. RBS and SEM analysis of the nickel-fullerene hybrid systems

    Czech Academy of Sciences Publication Activity Database

    Vacík, Jiří; Naramoto, J.; Narumi, K.; Yamanoto, S.; Abe, J.

    2003-01-01

    Roč. 206, - (2003), s. 395-398 ISSN 0168-583X Institutional research plan: CEZ:AV0Z1048901 Keywords : nicke-fullerene hybrid Subject RIV: BM - Solid Matter Physics ; Magnetism Impact factor: 1.041, year: 2003

  19. DNA replication: stalling a fork for imprinting and switching

    DEFF Research Database (Denmark)

    Egel, Richard

    2004-01-01

    Mating-type switching in fission yeast has long been known to be directed by a DNA 'imprint'. This imprint has now been firmly characterized as a protected site-specific and strand-specific nick. New work also links the widely conserved Swi1-Swi3 complex to the protection of stalled replication...

  20. Pop & rock / Valner Valme

    Index Scriptorium Estoniae

    Valme, Valner, 1970-

    2001-01-01

    Uute heliplaatide: Nick Cave And The Bad Seeds "No More Shall We Part", Andru Donalds "Let's Talk About It", No Angels "Elle'ments", The Experimental Pop Band "The Tracksuit Trilogy", Ellen ten Damme "I Am Here", Id_Rev "Sina ei" tutvustused

  1. Beyond Tokenism: Teaching a Diverse History in the Post-14 Curriculum

    Science.gov (United States)

    Dennis, Nick

    2016-01-01

    Nick Dennis shows how a "multidirectional memory" approach to teaching history can move history teachers beyond seeing black history as separate or distracting from the history that must be taught at examination level. He gives examples of ways in which a diverse history can be built into examination courses, strengthening historical…

  2. DNA expressions - A formal notation for DNA

    NARCIS (Netherlands)

    Vliet, Rudy van

    2015-01-01

    We describe a formal notation for DNA molecules that may contain nicks and gaps. The resulting DNA expressions denote formal DNA molecules. Different DNA expressions may denote the same molecule. Such DNA expressions are called equivalent. We examine which DNA expressions are minimal, which

  3. Fighting Fire with Fire

    DEFF Research Database (Denmark)

    Bossetta, Michael

    2017-01-01

    Advancing the concept of populism as a political style, this study compares the debate performances of two British party leaders, Nick Clegg and Nigel Farage, as they clashed in a pair of televised debates over Britain’s EU membership ahead of the 2014 European Parliament elections. The argument ...

  4. 76 FR 73608 - Reliability Technical Conference, North American Electric Reliability Corporation, Public Service...

    Science.gov (United States)

    2011-11-29

    ... or municipal authority play in forming your bulk power system reliability plans? b. Do you support..., North American Electric Reliability Corporation (NERC) Nick Akins, CEO of American Electric Power (AEP..., EL11-62-000] Reliability Technical Conference, North American Electric Reliability Corporation, Public...

  5. The Myth of Religious Experience Revisited

    Directory of Open Access Journals (Sweden)

    Zangwill Nick

    2017-10-01

    Full Text Available This article replies to Manuel Fasko’s “The Demystification of Nick Zangwill’s ‘Myth of Religious Experience’” (2017, showing how author’s argument against the possibility of religious experience presented in “The Myth of Religious Experience” (2004 remains in tact.

  6. The good, the bad, and the voter: the impact of hate speech prosecution of a politician on the electoral support for his party

    NARCIS (Netherlands)

    van Spanje, J.; de Vreese, C.

    2015-01-01

    Hate speech prosecution of politicians is a common phenomenon in established democracies. Examples of politicians tried for hate speech include Nick Griffin in Britain and Jean-Marie Le Pen in France. Does hate speech prosecution of politicians affect the electoral support for their party? This is

  7. Development Program for Field-Repairable/Expendable Main Rotor Blades

    Science.gov (United States)

    1976-09-01

    curve, based on the data obtained from this testing. In this curve, the runout points are plotted with arrows and the fatigue curve is drawn through...paper, or abrasive wheel , blend out scratch, nick, pitting, etc. CAUTION The final polish of smooth blending will be in a spanwise direction. Remove

  8. Enzyme-linked electrochemical DNA ligation assay using magnetic beads.

    Science.gov (United States)

    Stejskalová, Eva; Horáková, Petra; Vacek, Jan; Bowater, Richard P; Fojta, Miroslav

    2014-07-01

    DNA ligases are essential enzymes in all cells and have been proposed as targets for novel antibiotics. Efficient DNA ligase activity assays are thus required for applications in biomedical research. Here we present an enzyme-linked electrochemical assay based on two terminally tagged probes forming a nicked junction upon hybridization with a template DNA. Nicked DNA bearing a 5' biotin tag is immobilized on the surface of streptavidin-coated magnetic beads, and ligated product is detected via a 3' digoxigenin tag recognized by monoclonal antibody-alkaline phosphatase conjugate. Enzymatic conversion of napht-1-yl phosphate to napht-1-ol enables sensitive detection of the voltammetric signal on a pyrolytic graphite electrode. The technique was tested under optimal conditions and various situations limiting or precluding the ligation reaction (such as DNA substrates lacking 5'-phosphate or containing a base mismatch at the nick junction, or application of incompatible cofactor), and utilized for the analysis of the nick-joining activity of a range of recombinant Escherichia coli DNA ligase constructs. The novel technique provides a fast, versatile, specific, and sensitive electrochemical assay of DNA ligase activity.

  9. the efficacy of xylopia aethiopica and piper guinense seeds powder

    African Journals Online (AJOL)

    Sammy

    rice grain pests, it is essential that necessary control measures are put in place to ... Similarly, Teetes and Gilstrap (1986) were of the view that the use of chemical ... were killed, as suggested by Ivbijaro (1990) and Wee and Nick (1998).

  10. Zarqawi’s Sfumato: Operational Art in Irregular Warfare

    Science.gov (United States)

    2013-05-23

    in Baghdad and Karbala during the Shia festival of Ashura in March of 2004.80 These attacks were preceded by the bombing of the Jordanian embassy...American contractors Nick Berg and Eugene Armstrong. Zarqawi released edited videos with music and logos detailing multiple suicide bombings against

  11. The Significance of Bottom in Shakespeare's A Midsummer Night's Dream

    OpenAIRE

    KREMENIK, Michael

    1999-01-01

    The importance of the character Nick Bottom in A Midsummer Night's Dream will be discussed. He is the only character able to converse with both the natural and fairy worlds in the play. This makes his character special. This article will attempt to analyze why this is so.

  12. Pop / Mart Kuldkepp

    Index Scriptorium Estoniae

    Kuldkepp, Mart

    2008-01-01

    Heliplaatidest: The Wombats "A Guide to Love, Loss & Desperation", Lenny Kravitz "It is Time for a Love Revolution", Nick Cave & Warren Ellis "OST: The Assassination of Jesse James by the Coward Robert Ford", Pat Metheny "Secret Story", Lupe Fiasco "The Cool"

  13. A new protocol for extraction of C0t-1 DNA from rice

    African Journals Online (AJOL)

    USER

    2010-07-12

    Jul 12, 2010 ... plant species differentiation and genome evolution. A new protocol to ... According to nick translation theory, the genomic DNA was digested by DNase I and ... according to the formula Cot-1 = 1 = mol/L × Ts. Since highly and.

  14. 76 FR 56196 - Agency Information Collection Activities; Proposed Collection; Comment Request

    Science.gov (United States)

    2011-09-12

    ..., and IV); (b) the FTC's consumer complaint systems; (c) the FTC's program evaluation activities and (d... (Nick M) and Ami Dziekan (Ami D), Division of Enforcement, Bureau of Consumer Protection, Federal Trade... Commission's Rules of Practice (primarily Parts I, II, and IV); (b) the FTC's consumer complaint systems; (c...

  15. Wallace ja Gromit tulid Londonis linale / Andres Laasik

    Index Scriptorium Estoniae

    Laasik, Andres, 1960-2016

    2005-01-01

    Londonis esilinastus animafilm "Wallace ja Gromit : Libaküüliku needus" : režissöör Nick Park : Suurbritannia 2005. Kohal olid ka tegelastele hääle andnud Ralph Fiennes, Helena Bonham Carter ja Peter Sallis

  16. The Pedagogue of the Auratic Medium--Extending the Argument

    Science.gov (United States)

    Dobson, Stephen

    2008-01-01

    Nick Peim has recently revisited the work of Walter Benjamin; specifically his famous essay on art and mechanical reproduction. In this reply, I too draw upon the inspiration of Benjamin to extend the argument to the question of experience and what might count as knowledge, both in a philosophical sense and also in terms of the curriculum. To…

  17. Analysis of SSN 688 Class Submarine Maintenance Delays

    Science.gov (United States)

    2017-06-01

    Simplified Notional Submarine FRP (Independent Deployer) ..................11  Figure 8.  Evolution of Los Angeles Class Submarine Notional...Number TFP Technical Foundation Paper URO Unrestricted Operations xv ACKNOWLEDGMENTS I would like to thank my lead advisor, Professor Nick Dew...only on Los Angeles (SSN 688)-class submarines. Being the higher quantity and older generation submarine hull type, the Los Angeles class submarine

  18. Children in Hot Cars Result in Fatal Consequences

    Medline Plus

    Full Text Available ... a hot vehicle. “Putting it bluntly, leaving your child in a hot car is like leaving your child in a lit oven,” said Dr. Nick Jouriles. “ ... or experience changes in your schedule or your child is in the back seat, making it more ...

  19. Limited numbers of apoptotic cells in fresh paraffin embedded bone marrow samples of patients with myelodysplastic syndrome

    NARCIS (Netherlands)

    Brada, SJL; van de Loosdrecht, AA; Koudstaal, J; de Wolf, JTM; Vellenga, E

    In myelodysplasia (MDS) the precise mechanism of ineffective erythropoiesis is not fully elucidated, but it is suggested that apoptosis may contribute to this process. We performed TdT-mediated dUTP-nick end labelling (TUNEL) staining of paraffin embedded bone marrow specimens to assess the amount

  20. Before Coffee, Facebook: New Literacy Learning for 21st Century Teachers

    Science.gov (United States)

    Roach, Audra K.; Beck, Jessica J.

    2012-01-01

    And so a middle school language arts teacher begins her Saturday. Before coffee, Facebook. In this themed issue on professional development in an era of nick.com, the authors propose that teachers' new literacy learning is as close as their own screens. Teachers, too, live literate lives online in this new age of composition, with impulses to…

  1. Berlinale terad / Kadri Kõusaar

    Index Scriptorium Estoniae

    Kõusaar, Kadri

    2006-01-01

    56. Berliini festivali filme - Jasmila Zhbanici "Grbavica", Michael Winterbottomi, Mat Whitecrossi "Teekond Guantanamosse", tailase Pen-ek Ratanaruangi "Nähtamatud lained", John Hillcoati "Ettepanek" (stsenarist ja muusika autor Nick Cave), Bennett Milleri "Capote" (nimiosas P.S. Hoffman), Yang Yong-hi dokfilm "Kallis Pyongyang", Tomer Heymanni dokfilm "Pabernukud"

  2. 77 FR 8892 - Detailed Planning To Consider Additional Land Protection on the Missouri River From Fort Randall...

    Science.gov (United States)

    2012-02-15

    ... 25486, DFC, Denver, CO 80225. FOR FURTHER INFORMATION CONTACT: Nick Kaczor, Planning Team Leader... old, wider river valley contains important habitat for at least 60 native and 26 sport fish. In... historical characteristics witnessed by Lewis and Clark during their explorations in the early 1800s. The FWS...

  3. Student Engagement, Ideological Contest and Elective Affinity: The Zepke Thesis Reviewed

    Science.gov (United States)

    Trowler, Paul

    2015-01-01

    This paper takes up issues raised in two articles by Nick Zepke and portrayed here as "the Zepke thesis". This thesis argues that the literature on, interest in and practices around student engagement in higher education have an elective affinity with neo-liberal ideology. At one level this paper counters many of the assertions that…

  4. Fulltext PDF

    Indian Academy of Sciences (India)

    In the first group, the paper by Nick Devereux reviews Hubble Space .... here that the STARK-B database is included in the new search facility for such data namely Virtual Atomic and Molecular Data Center (VAMDC – http://vamdc.org/).

  5. Briti publik armastab King Kongi / Andris Feldmanis

    Index Scriptorium Estoniae

    Feldmanis, Andris, 1982-

    2006-01-01

    Briti filmiajakiri Empire lugejate filmiauhinnad : parim film "King Kong" (Peter Jackson), parim režii Steve Box, Nick Park "Wallace & Gromit - libaküüliku needus", parim meesnäitleja Johnny Depp, parim naisnäitleja Thandie Newton

  6. Turism rikastab : rahvusvaheline turismikonverents

    Index Scriptorium Estoniae

    2006-01-01

    28. septembril toimus Tallinnas Rahvusraamatukogus rahvusvaheline turismikonverents"Turism rikastab". Ülevaade majandus- ja kommunikatsiooniministeeriumi majandusarengu asekantsleri Maria Värtoni, Graham Mckenzie, Linnar Viigi, Nick Skelloni, Mario Ascencao (Haagi ülikool), David Carsoni (Ulsteri ülikool), Vesa Heikkineni ja Imre Sooääre ettekannetest

  7. Moskvas hoitav Adolf Hitleri kolbatükk ei kuulu füürerile / Aadu Hiietamm

    Index Scriptorium Estoniae

    Hiietamm, Aadu, 1954-

    2009-01-01

    USA arheoloogi Nick Bellantoni avastus, et Moskvas hoitav Adolf Hitleri kuuliauguga koljutükk ei kuulugi füürerile, annab tõenäoliselt uut hoogu vandenõuteoreetikutele, kes usuvad, et Hitler ei teinud 1945. aastal enesetappu, vaid põgenes välismaale

  8. Tänavuse Berlinale filmid süvenevad ihulähedasse / Grete Nootre

    Index Scriptorium Estoniae

    Nootre, Grete

    2006-01-01

    56. Berliini filmifestivali programmidest ja filmidest - USA mängufilm "A Prairie Home Companion" : režissöör Robert Altman, saksa mängufilm "Vaba tahe" ("Der Freie Wille") : režissöör Matthias Glasner, austraalia-inglise "Ettepanek" ("The Proposition") : stsenarist ja helilooja Nick Cave : režissöör John Hillcoat

  9. Muusika / Mart Juur

    Index Scriptorium Estoniae

    Juur, Mart, 1964-

    2005-01-01

    Uutest heliplaatidest Pia Fraus "Sailing On A Grapefruit Lake", Six Organs Of Admittance "School Of The Flower", Jaga Jazzist "What We Must", Oasis "Don't Belive The Truth", Limp Bizkit "The Unquestionable Truth", "Do Me Bad Things", Saari, Jarmo "Solu", Cave, Nick & The Bad Seeds "B-Sides & Rarities", "Stars Wars", Weezer "Make Believe"

  10. Muusika DVD-d / Priit Hõbemägi

    Index Scriptorium Estoniae

    Hõbemägi, Priit, 1957-

    2006-01-01

    Uutest DVD-dest Moody Blues "Lovely to See You - Live", Nick Cave and The Bad Seeds "Road to God Knows Wherw", Thin Lizzy "Thunder and Lightning Tour", Eels "Eels With Strings - Live at Town Hall", Moby "Live. Hotel Tour 2005", Corvus Corax "Cantus Buranus", Destiny's Child "Live in Atlanta"

  11. Äraigatsuse salakajad / Aita Kivi

    Index Scriptorium Estoniae

    Kivi, Aita, 1954-

    2001-01-01

    Sisu : Rein Põder. Hula; David Shahar. Tema majesteedi agent; Stephen Kuusisto. Pimedate planeet; Joanna Trollope. Mehed ja tütarlapsed; Detlef Jena. Venemaa tsaarinnad; Helle Helle. Maja ja kodu; Nick Hornby. Ühest poisist; Deepak Chopra. Eatu keha, ajatu vaim

  12. Remember the Algae that Went to Space? Here's What Happened Next | News |

    Science.gov (United States)

    . He met with Dr. Alexandra Dubini, at the time a research scientist at NREL working with the algae education how to grow algae. Nick Sweeney, a research technician in NREL's National Bioenergy Center . vulgaris to shift into lipid production mode. Squares of a special tape that changes color from tan to gray

  13. Plaadid / Villu Päärt

    Index Scriptorium Estoniae

    Päärt, Villu, 1972-

    2002-01-01

    Uutest plaatidest Nick Carter "Now Or Never", Brian Mcknight "From There To here 1989-2002", Rod Stuart "The Grat American Songbook", New Order "International", Justin Timberlake "Justified", Manic Street Prachers "Forever Delayed - Greatest Hits", LHB "Tell 'Em Who We Are"

  14. Work-Life Balance: the Links with Management Practices and Productivity

    OpenAIRE

    Nick Bloom; Tobias Kretschmer; John Van Reenen

    2006-01-01

    Does good management and higher productivity come at the expense of work-life balance? Or is good work-life balance an important component of the management of successful firms? New research by Nick Bloom, Tobias Kretschmer and John Van Reenen finds evidence for a hybrid view between these two polar extremes.

  15. Landscapes of The Mind: The Setting as Psyche in Nathaniel Hawthorne's The Scarlet Letter And F. Scott Fitzgerald's The Great Gatsby Landscapes of The Mind: The Setting as Psyche in Nathaniel Hawthorne's The Scarlet Letter And F. Scott Fitzgerald's The Great Gatsby

    Directory of Open Access Journals (Sweden)

    Cruce Stark

    2009-09-01

    Full Text Available In the closing moment of F. Scott Fitzgerald's The Great Gateby, Nick returns for one last look at what once was Gateby's house. But instead of physical buildings, he has a vision of an earlier time, a vision of the "old island" that "flowered once for Dutch sailor's eyes -- a fresh green breast of the new world." "For a transitory enchanted moment," Nick thinks, "man must have held his breath in the presence of this continent, compelled into an aesthetic contemplation he neither understood nor desired, face to face for the last time in history with something commensurate to his capacity for wonder (182." In the closing moment of F. Scott Fitzgerald's The Great Gateby, Nick returns for one last look at what once was Gateby's house. But instead of physical buildings, he has a vision of an earlier time, a vision of the "old island" that "flowered once for Dutch sailor's eyes -- a fresh green breast of the new world." "For a transitory enchanted moment," Nick thinks, "man must have held his breath in the presence of this continent, compelled into an aesthetic contemplation he neither understood nor desired, face to face for the last time in history with something commensurate to his capacity for wonder (182."

  16. 76 FR 36903 - Privacy Act of 1974; System of Records

    Science.gov (United States)

    2011-06-23

    ... have a need-to-know. Individuals responsible for servicing the records in the performance of official... name, nick names, Social Security Number (SSN), gender, date of birth, personal cell phone number, home... address: Delete entry and replace with ``Automated Civil Engineer System/ Interim Work Management System...

  17. Wallace ja Gromit tegid Hollywoodis puhta töö / Andris Feldmanis

    Index Scriptorium Estoniae

    Feldmanis, Andris, 1982-

    2006-01-01

    Hollywoodi animafilmide Annie-auhindade jagamisel võitis 9 auhinda täispikk animafilm "Wallace ja Gromit : Libaküüliku needus" ("Wallace & Gromit : The Curse of the Were-Rabbit") : režissöörid Nick Park ja Steve Box : Suurbritannia 2005

  18. Employment Relations in Denmark

    DEFF Research Database (Denmark)

    Madsen, Jørgen Steen; Due, Jesper Jørgen; Andersen, Søren Kaj

    2011-01-01

    Jørgen Steen Madsen, Jesper Due og Søren Kaj Andersen har skrevet et kapitel om udviklingen i dansk arbejdsmarkedsregulering til bogen International and Comparative Employment Relations, redigeret af Greg Bamber, Russell Lansbury og Nick Wailes. Bogen indeholder bidrag, der præsenterer og...

  19. Recent Naval Postgraduate School Publications

    Science.gov (United States)

    1988-09-30

    pay, performance ratings and voluntary separations 12th Annual Convention, Eastern Econ. Asso., Philadelphia, PA, Apr., 1986. Sol nick, L M...ilow above the Marine boundary layer 4th Conference Meteorol. and Oceano . Coastal Zone, Anaheim, CA., February, 3-5, 1988. Smith, R Computational

  20. "Of comics and men: A cultural history of American comic books," by Jean-Paul Gabilliet

    Directory of Open Access Journals (Sweden)

    Drew Morton

    2013-06-01

    Full Text Available Jean-Paul Gabilliet. Of comics and men: A cultural history of American comic books. Translated by Bart Beaty and Nick Nguyen. Jackson: University Press of Mississippi, 2010, hardcover, $55.00 (390p, ISBN 978-1604732672.

  1. Uudised

    Index Scriptorium Estoniae

    2005-01-01

    Eesti artisti Pafi ehk Paavo Eensalu lugu "Daisy" ilmub septembris Nick Luscombe poolt kokkupandaval kogumikul "Flo-Motion vol 2". Popansambel Radiohead alustas oma uut LPd tehes blogi ehk internetipäeviku "Dead Air Space" pidamist, aadressilt http://www.radiohead.com/deadairspace/ võib leida bändiliikmete kirjutisi

  2. Research Staff | Photovoltaic Research | NREL

    Science.gov (United States)

    Research Staff Research Staff desc Greg Wilson Center Director Dr. Greg Wilson is the Director of @nrel.gov 303-384-6649 Bosco, Nicholas Staff Scientist Nick.Bosco@nrel.gov 303-384-6337 Braunecker, Wade IV-Physics Michael.Deceglie@nrel.gov 303-384-6104 Deline, Chris Staff Engineer Chris.Deline@nrel.gov

  3. Ostrich: Journal of African Ornithology - Vol 88, No 1 (2017)

    African Journals Online (AJOL)

    DOWNLOAD FULL TEXT DOWNLOAD FULL TEXT. Abdulmaula Hamza, Nicola Baccetti, Joe Sultana, Jaber Yahia, Marco Zantello, Adriano de Faveri, Nick Cutts, John Borg, Hichem Azafzaf, Pierre Defos du Rau, Esam Bourass, Khaled Etayeb, Michael Elliott, 53–58. http://dx.doi.org/10.2989/00306525.2016.1231139 ...

  4. CNS wound healing is severely depressed in metallothionein I- and II-deficient mice

    DEFF Research Database (Denmark)

    Penkowa, M; Carrasco, J; Giralt, M

    1999-01-01

    . In contrast to normal mice, at 20 dpl no wound healing had occurred. The rate of apoptosis, as determined by using terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling, was drastically increased in neurons of ipsilateral cortex of the MT-I+II null mice. Our results demonstrate that MT...

  5. Healthcare reform. Is the NHS ready for US business guru's strategy?

    Science.gov (United States)

    Cavendish, Will; Edwards, Nigel; Swindells, Matthew; Henke, Nicolaus; Robinson, Edna; Smith, Richard

    2006-12-07

    The central argument of the new book by renowned US academics Michael Porter and Elizabeth Olmsted Teisberg is that the US health system is broken because rather than improving quality and efficiency, it focuses on budgetary battles. HSJ gathered together six leading healthcare insiders to discuss whether his diagnosis is applicable to the NHS. Nick Edwards was there.

  6. Ganoderic acid B's influence towards the therapeutic window of ...

    African Journals Online (AJOL)

    9. Hempel C, Norenberg W, Sobottka H, Urban N, Nicke. A, Fischer W, Schaefer M. The phenothiazine-class an- tipsychotic drugs prochlorperazine and trifluoperazine are potent allosteric modulators of the human P2X7 receptor. Neuropharmacology 2013; 75:365-379. 10. Song H, Tao Y, Zhang CZ, Zhou YD. Comparison.

  7. Uus peaminister, uudne valitsus / Kaivo Kopli

    Index Scriptorium Estoniae

    Kopli, Kaivo

    2010-01-01

    Suurbritannia sai koalitsioonivalitsuse. Nii peaminister David Cameron kui ka asepeaminister Nick Clegg on vihjanud, et eelistavad lõdvemaid suhteid USA-ga, kui need olid Gordon Brownil ja Tony Blairil. Konservatiivide partei on skeptiline EL-i koostöö suhtes

  8. The Guatemalan Military: Transition from War to Peace

    Science.gov (United States)

    2001-06-01

    1992. Cerdas Cruz, Rodolfo. El Precio de Una Herencia. Fuerza Armadas, Democracia y Derechos Humanos en Centroamérica. San José: Instituto...Interamericano de Derechos Humanos , 1996. Cullather, Nick. Secret History, The CIA’s Classified Account of Its Operations in Guatemala, 1952-1954. Stanford

  9. Storage stability of flour-blasted brown rice

    Science.gov (United States)

    Brown rice was blasted with rice flour rather than sand in a sand blaster to make microscopic nicks and cuts so that water can easily penetrate into the brown rice endosperm and cook the rice in a shorter time. The flour-blasted American Basmati brown rice, long grain brown rice, and parboiled long...

  10. Cognitive Psychology and Low-Stakes Testing without Guarantees

    Science.gov (United States)

    Dennis, Nick

    2016-01-01

    The emphasis on the power of secure substantive knowledge reflected in recent curriculum reforms has prompted considerable interest in strategies to help students retain and deploy such knowledge effectively. One strategy that has been strongly endorsed by some cognitive psychologists is regular testing; an idea that Nick Dennis set out to test…

  11. Adenosine triphosphate stimulates Aquifex aeolicus MutL endonuclease activity.

    Directory of Open Access Journals (Sweden)

    Jerome Mauris

    2009-09-01

    Full Text Available Human PMS2 (hPMS2 homologues act to nick 5' and 3' to misincorporated nucleotides during mismatch repair in organisms that lack MutH. Mn(++ was previously found to stimulate the endonuclease activity of these homologues. ATP was required for the nicking activity of hPMS2 and yPMS1, but was reported to inhibit bacterial MutL proteins from Thermus thermophilus and Aquifex aeolicus that displayed homology to hPMS2. Mutational analysis has identified the DQHA(X(2E(X(4E motif present in the C-terminus of PMS2 homologues as important for endonuclease activity.We examined the effect ATP had on the Mn(++ induced nicking of supercoiled pBR322 by full-length and mutant A. aeolicus MutL (Aae MutL proteins. Assays were single time point, enzyme titration experiments or reaction time courses. The maximum velocity for MutL nicking was determined to be 1.6+/-0.08x10(-5 s(-1 and 4.2+/-0.3x10(-5 s(-1 in the absence and presence of ATP, respectively. AMPPNP stimulated the nicking activity to a similar extent as ATP. A truncated Aae MutL protein composed of only the C-terminal 123 amino acid residues was found to nick supercoiled DNA. Furthermore, mutations in the conserved C-terminal DQHA(X(2E(X(4E and CPHGRP motifs were shown to abolish Aae MutL endonuclease activity.ATP stimulated the Mn(++ induced endonuclease activity of Aae MutL. Experiments utilizing AMPPNP implied that the stimulation did not require ATP hydrolysis. A mutation in the DQHA(X(2E(X(4E motif of Aae MutL further supported the role of this region in endonclease activity. For the first time, to our knowledge, we demonstrate that changing the histidine residue in the conserved CPHGRP motif abolishes endonucleolytic activity of a hPMS2 homologue. Finally, the C-terminal 123 amino acid residues of Aae MutL were sufficient to display Mn(++ induced nicking activity.

  12. Structure-guided mutational analysis of the OB, HhH, and BRCT domains of Escherichia coli DNA ligase.

    Science.gov (United States)

    Wang, Li Kai; Nair, Pravin A; Shuman, Stewart

    2008-08-22

    NAD(+)-dependent DNA ligases (LigAs) are ubiquitous in bacteria and essential for growth. LigA enzymes have a modular structure in which a central catalytic core composed of nucleotidyltransferase and oligonucleotide-binding (OB) domains is linked via a tetracysteine zinc finger to distal helix-hairpin-helix (HhH) and BRCT (BRCA1-like C-terminal) domains. The OB and HhH domains contribute prominently to the protein clamp formed by LigA around nicked duplex DNA. Here we conducted a structure-function analysis of the OB and HhH domains of Escherichia coli LigA by alanine scanning and conservative substitutions, entailing 43 mutations at 22 amino acids. We thereby identified essential functional groups in the OB domain that engage the DNA phosphodiester backbone flanking the nick (Arg(333)); penetrate the minor grove and distort the nick (Val(383) and Ile(384)); or stabilize the OB fold (Arg(379)). The essential constituents of the HhH domain include: four glycines (Gly(455), Gly(489), Gly(521), Gly(553)), which bind the phosphate backbone across the minor groove at the outer margins of the LigA-DNA interface; Arg(487), which penetrates the minor groove at the outer margin on the 3 (R)-OH side of the nick; and Arg(446), which promotes protein clamp formation via contacts to the nucleotidyltransferase domain. We find that the BRCT domain is required in its entirety for effective nick sealing and AMP-dependent supercoil relaxation.

  13. Ecological consequences of anthropogenic pressure in Wari-Maro Forest Reserve (Benin, West Africa

    Directory of Open Access Journals (Sweden)

    Aubin Guénolé Amagnide

    2015-12-01

    Full Text Available This study assessed ecological consequences of anthropogenic pressure on Wari-Maro Forest Reserve (WMFR. The dynamics of forest cover has been assessed using a diachronic analysis of land cover maps from the Landsat satellite images of 1986, 1995 and 2006. Structural patterns of the forest has been described using forest inventory data with twenty five 1ha plots having two 50 m x 30 m plots set up inside and positioned at the opposite corners of the leading diagonal within each 1 ha plot. Established plots allowed identifying the most targeted species in illegal logging. Plots of 0.15 ha established inside each 1 ha plot helped assessing the volume of trees from which we derived carbon stock and carbon loss using conversion and expansion factors. For the two periods 1986 to 1995 and 1995 to 2006, there was a decline in forest cover which slowed down in the second decade (0.196 %.year-1 and 0.083 %.year-1 respectively. The two vegetation types of the WMFR were mainly distinguished by Lorey's mean height (12.81 m in woodland and 12.44 m in tree-savannah. Top five targeted species in illegal logging activities were: Pterocarpus erinaceus Poir., Afzelia africana Sm., Isoberlinia spp., Anogeissus leiocarpa Guill. and Daniellia oliveri (Rolfe Hutch. & Dalziel. Results also showed mean values of carbon stock and carbon losses for the whole forest of 147.84 tons C.ha-1 and 17.57 tons C.ha-1 respectively and did not depend on vegetation type. Results from this study suggest that management strategies should focus on selectively logged species. Monitoring should also be enhanced to ensure conservation of resources of the reserve which are at high risks of extinction due to selective logging rates. Keywords: anthropogenic pressure, forest cover, structure, carbon stock, Wari-Maro forest reserve, Benin.

  14. Diversité, priorité pastorale et de conservation des ligneux fourragers des terres de parcours en zone soudano-guinéenne du Bénin

    Directory of Open Access Journals (Sweden)

    Sèwadé, C.

    2016-01-01

    Full Text Available Diversity, pastoral and conservation priorities of fodder trees in the Sudano-Guinean pasture lands of Benin. Description of the subject. Fodder trees are important for livestock survival in dryland Africa. In view of the pressure faced by these trees, and their consequent rarity noted in rangelands, a study was conducted in the Sudano-Guinean transition zone of Benin at the level of the local population surrounding the protected forests of Monts Kouffé, Wari-Maro and Ouémé Supérieur. Objectives. This study aimed to inventory the fodder trees, analyze the local perception of factors threatening target fodder trees, according to different sociolinguistic groups and prioritize fodder trees for conservation. Method. Ethnobiological surveys and ecological data from the available literature were used to construct a database following different criteria. The citation rates of the fodder trees by the surveyed populations were used to establish pastoral priority, while their conservation priority was established using a combination of four methods and nine criteria. Results. Forty-eight fodder trees belonging to 17 families dominated by Leguminosae (27.1% and Moraceae (16.6% were reported. These species were distributed among 37 genera, with the genus Ficus being the most represented (16.6%. Palatability, species availability and the impact of tree fodder on animal productivity were the criteria used by the surveyed sociolinguistic groups in their selection of fodder trees. The prioritization methods yielded ten top ranked species: Afzelia africana, Pterocarpus erinaceus, Khaya senegalensis, Vitellaria paradoxa, Mangifera indica, Ficus platyphylla, Balanites aegyptiaca, Annona senegalensis, Ficus umbellata and Daniellia oliveri. Conclusions. With the aim of establishing the sustainable management of pasture lands, we suggest that priority be given to the aforementioned species of fodder trees as part of restoration, afforestation/reforestation and

  15. On the identity of the adventive species of Eufriesea Cockerell in the USA: systematics and potential distribution of the coerulescens species group (Hymenoptera, Apidae

    Directory of Open Access Journals (Sweden)

    Victor H. Gonzalez

    2017-04-01

    Full Text Available In the summer of 2010, two male specimens of the neotropical orchid bee genus Eufriesea Cockerell were collected in the Guadalupe Mountains of western Texas and southeastern New Mexico, USA. We tentatively identified them as E. coerulescens (Lepeletier de Saint Fargeau because of the uncertainty surrounding the limits of this taxon and hypothesized that they were members of a persistent bee population, rather than long-distance transient vagrants. The goals of this paper are to clarify the identity of these specimens, assess the species limits of E. coerulescens, and to evaluate suitability of habitats in the USA for this adventive species. Herein, we revise the species in the coerulescens group using morphological features of both sexes and confirm that the specimens of Eufriesea from the USA are E. coerulescens. We recognize the following six species in the coerulescens group: E. coerulescens, E. micheneri Ayala & Engel, E. simillima (Moure & Michener, which is reinstated from synonym with E. coerulescens, and three new species from Mexico (E. barthelli Gonzalez & Griswold, sp. n., E. engeli Gonzalez & Griswold, sp. n., and E. oliveri Gonzalez & Griswold, sp. n.. To facilitate the identification of these taxa, we present a fully illustrated account of the species, comparative diagnoses, descriptions, and an updated key to all Mexican species of Eufriesea. Our analyses using species distribution modelling show an absence of suitable habitat for E. coerulescens in western Texas and southeastern New Mexico, thus favoring the long-distance dispersal hypothesis. The analyses also suggest high suitability of habitats across the Caribbean and some areas in Florida, as well as in other regions in Mexico and Central America. We discuss the implications of these results and compare them with the predicted distribution available for the other two known adventive orchid bee species in the USA.

  16. The effect of irradiation on the DNA of cauliflower

    International Nuclear Information System (INIS)

    Harmey, M.A.

    1991-01-01

    The cellular DNA is one of the components most affected by ionizing radiation. Lesions caused range from single and double stranded breaks to chemical modification of bases depending on the radiation dosage and the metabolic status of the tissue. In attempting to assess the DNA damage induced by irradiation of vegetables in a speedy and convenient manner, we examined the effect on the DNA by subjecting cauliflower to a dose of 1 kGy. If DNA is nicked by irradiation, the extent of the damage can be assessed by using DNA polymerase to repair the nicks. Comparisons were made between irradiated and non irradiated cauliflower and incorporation of 32 p deoxy GTP in the presence of the Klenow fragment of DNA polymerase measured

  17. Naval Surface Forces Real-Time Reutilization Asset Management Warehouses: A Cost-Benefit Analysis

    Science.gov (United States)

    2008-12-01

    shared their valuable insights. And finally, to our advisors Dr . Ken Euske and CDR Brett Wagner who kept us on track throughout the project. Nick could...of $206,368,657. SURFOR’s East and West Coast warehouses accounted for $146,975,108 of the list value. The team identified potential cost...obligating the funds. A relatively simple cost benefit analysis of the reprogramming costs versus the cost savings would justify the expense. In the

  18. Strategic Reflections: Operation Iraqi Freedom, July 2004-February 2007

    Science.gov (United States)

    2012-10-01

    not pan out. We knew that our mission was ultimately to hand over security responsibility to the Iraqi government, but we had not yet developed a...were “ chicken -egg” discus- sions about whether the security situation had to improve before the political track could begin. I strongly argued that...2005 Lt Gen Nick Houghton (UK), October 2005–February 2006 Lt Gen Rob Fry (UK), March 2006–September 2006 Lt Gen Graham Lamb (UK), September 2006

  19. Ian Curtis (2001)

    OpenAIRE

    Dalwood, D

    2016-01-01

    This exhibition offers a presentation of FRAC Champagne-Ardenne‘s collection; artists included: Francesco Arena, Sylvie Auvray, Dara Birnbaum, Rossella Biscotti, Tom Burr, Antoine Catala, Guy de Cointet, Ann Craven, Dexter Dalwood, Hassan Darsi, Plamen Dejanoff, Marcelline Delbecq, Julien Discrit, Jimmie Durham, Ruth Ewan, Michel François, Linda Fregni Nagler, Aurélien Froment, General Idea, Oscar Giaconia, Jean-Michel Hannecart, Sharon Hayes, Charline von Heyl, Barbara Kasten, Nick Mauss, Ma...

  20. Can this merger be saved?

    Science.gov (United States)

    Cliffe, S

    1999-01-01

    In this fictional case study, a merger that looked like a marriage made in heaven to those at corporate headquarters is feeling like an infernal union to those on the ground. The merger is between Synergon Capital, a U.S. financial-services behemoth, and Beauchamp, Becker & Company, a venerable British financial-services company with strong profits and an extraordinarily loyal client base of wealthy individuals. Beauchamp also boasts a strong group of senior managers led by Julian Mansfield, a highly cultured and beloved patriarch who personifies all that's good about the company. Synergon isn't accustomed to acquiring such companies. It usually encircles a poorly managed turnaround candidate and then, once the deal is done, drops a neutron bomb on it, leaving file cabinets and contracts but no people. Before acquiring Beauchamp, Synergon's macho men offered loud assurances that they would leave the tradition-bound company alone-provided, of course, that Beauchamp met the ambitious target numbers and showed sufficient enthusiasm for cross-selling Synergon's products to its wealthy clients. In charge of making the acquisition work is Nick Cunningham, one of Synergon's more thoughtful executives. Nick, who was against the deal from the start, is the face and voice of Synergon for Julian Mansfield. And Mansfield, in his restrained way, is angry at the constant flow of bureaucratic forms, at the rude demands for instant information, at the peremptory changes. He's even dropping broad hints at retirement. Nick has already been warned: if Mansfield goes, you go. Six commentators advise Nick on how to save his job by bringing peace and prosperity to the feuding couple.

  1. Briefing paper for universities on Core Maths

    OpenAIRE

    Glaister, Paul

    2015-01-01

    This briefing paper outlines the rationale for and development of the new Core Maths qualifications, the characteristics of Core Maths, and why Core Maths is important for higher education. It is part of a communication to university vice-chancellors from the Department for Business, Innovation and Skills (BIS) comprising this paper and a joint Ministerial letter from Jo Johnson, Minister of State for Universities and Science in BIS, and Nick Gibb, Minister of State for Schools in the Departm...

  2. Environmental tobacco smoke is just as damaging to DNA as mainstream smoke.

    OpenAIRE

    Bermúdez, E; Stone, K; Carter, K M; Pryor, W A

    1994-01-01

    This study demonstrates the ability of tar isolated from environmental tobacco smoke (ETS) to nick DNA in mammalian cells. Solutions of ETS tar behave similarly to aqueous solutions of cigarette tar from mainstream smoke. Both solutions contain the tar semiquinone radical, and this radical associates with the DNA in viable rat alveolar macrophages. Solutions of tar from ETS cause single-strand DNA breaks in rat thymocytes in proportion to the amount of tar present, until a plateau is reached....

  3. Realismist animatsioonfilmis / Ülo Pikkov

    Index Scriptorium Estoniae

    Pikkov, Ülo, 1976-

    2007-01-01

    Animafilmist kui realistlikust filmiliigist. Artiklis viited ka mitmetele filmiteoreetikutele nagu Juri Lotman, Martin Heidegger, Steve Reinke, Paul Wells, Stephen Rowley. Näited filmidest "Lusitania uppumine" ("The Sinking of theLusitania", Winsor McCay, 1918), "Ryan" (Chris Landherth, 2004), "Creature Comforts" (Nick Park, 1989), "Pocahontas" (1995), "Jurassic Park" (Steven Spielberg, 1993), "Beavis ja Butt-head" (1993-1997), "South Park" (1997-2007)

  4. JPRS Report, East Europe.

    Science.gov (United States)

    1990-01-17

    21 Oct] ............................................. 38 THM Dal Plans Clothing Goods Store Franchises [RYNKI ZAGRANICZNE 7 Oct...former classmate Radomir Mladenovic, nick- General Dobrnjac was supposed to have lived. All we named Duce, the owner of the Boter cafe . had left was a... cafe . I know that he would stayed with their mother in Pozarevac. Slobodan not like it, even though he is an idol for many people Milosevic’s father

  5. Psychological and social analysis of population of Goiania three years after the accident with the cesium-137

    International Nuclear Information System (INIS)

    Helou, Suzana; Costa Neto, Sebastiao Benicio da; Curado, Maria Paula

    1995-01-01

    Psychological and social analysis of population of Goiania - Brazilian city - three years after the accident with the cesium-137 occurred in 1987 are discussed. With this goal it is presented a public opinion research in order to retract the Goiania's radioactive accident residual psychological effects. The public opinion research was analyzed in sum individual answers terms, considering the factors multiplicity to what the individuals are exposed in the society. It was adopted Cabral's and Nick's concepts

  6. WannaBike - one of the teams that participated in Bike2Work 2017

    CERN Multimedia

    2017-01-01

    Bike2Work is a healthy living initiative involving companies across Switzerland, and in May and June this year it inspired 54 780 participants from 1 885 companies to take to the saddle for their daily commute, while simultaneously promoting a sustainable approach to transport. In the photo one can see the team "WannaBike" from CERN. The members of the team, from left to right, were: Nick Walter, Ludivine Ceard, Cecilia Uribe Estrada and Manuel Silva.

  7. Colliding muons

    International Nuclear Information System (INIS)

    Anon.

    1994-01-01

    Is a muon-muon collider really practical? That is the question being asked by Bob Palmer. Well known in particle physics, Palmer, with Nick Samios and Ralph Shutt, recently won the American Physical Society's Panofsky Prize for their 1964 discovery of the omega minus. As well as contributing to other major experiments, both at CERN and in the US, he has contributed ideas to stochastic cooling and novel acceleration schemes

  8. Still a Special Relationship? The Significance of United States-United Kingdom Relations in the Twenty-First Century

    Science.gov (United States)

    2011-12-01

    Anglo - Saxon ally over Nasser and the canal.55 In 2010, in a speech at Chatham House, Liberal Democrat Nick Clegg suggested that “what...strategic culture, and as an expression of shared values and institutions the character of which is vital for those charged with service in an Anglo ...American context as well as NATO. 14. SUBJECT TERMS United Kingdom, United States, Special Relationship, Anglo - American Relations, Iraq, Afghanistan

  9. Exploring the Molecular Mechanisms of Nickel-Induced Genotoxicity and Carcinogenicity: A Literature Review

    OpenAIRE

    Cameron, Keyuna S.; Buchner, Virginia; Tchounwou, Paul B.

    2011-01-01

    Nickel, a naturally occurring element that exists in various mineral forms, is mainly found in soil and sediment, and its mobilization is influenced by the physicochemical properties of the soil. Industrial sources of nickel include metallurgical processes such as electroplating, alloy production, stainless steel, and nickel-cadmium batteries. Nickel industries, oil- and coal-burning power plants, and trash incinerators have been implicated in its release into the environment. In humans, nick...

  10. Drug and Alcohol Studies (Volume 5: Interventions)

    OpenAIRE

    MacGregor, S; Thom, B

    2014-01-01

    VOLUME FIVE: INTERVENTIONS Natural Recovery from Alcohol Problems Harald Klingemann School-Based Programmes to Prevent Alcohol, Tobacco and Other Drug Use Gilbert Botvin and Kenneth Griffin Community Prevention of Alcohol Problems Harold Holder Can Screening and Brief Intervention Lead to Population-Level Reductions in Alcohol-Related Harm? Nick Heather Sharpening the Focus of Alcohol Policy from Aggregate Consumption to Harm and Risk Reduction Tim Stockwell et al A Review of the Efficacy and...

  11. Functional properties and structural requirements of the plasmid pMV158-encoded MobM relaxase domain.

    Science.gov (United States)

    Fernández-López, Cris; Pluta, Radoslaw; Pérez-Luque, Rosa; Rodríguez-González, Lorena; Espinosa, Manuel; Coll, Miquel; Lorenzo-Díaz, Fabián; Boer, D Roeland

    2013-07-01

    A crucial element in the horizontal transfer of mobilizable and conjugative plasmids is the relaxase, a single-stranded endonuclease that nicks the origin of transfer (oriT) of the plasmid DNA. The relaxase of the pMV158 mobilizable plasmid is MobM (494 residues). In solution, MobM forms a dimer through its C-terminal domain, which is proposed to anchor the protein to the cell membrane and to participate in type 4 secretion system (T4SS) protein-protein interactions. In order to gain a deeper insight into the structural MobM requirements for efficient DNA catalysis, we studied two endonuclease domain variants that include the first 199 or 243 amino acid residues (MobMN199 and MobMN243, respectively). Our results confirmed that the two proteins behaved as monomers in solution. Interestingly, MobMN243 relaxed supercoiled DNA and cleaved single-stranded oligonucleotides harboring oriTpMV158, whereas MobMN199 was active only on supercoiled DNA. Protein stability studies using gel electrophoresis and mass spectrometry showed increased susceptibility to degradation at the domain boundary between the N- and C-terminal domains, suggesting that the domains change their relative orientation upon DNA binding. Overall, these results demonstrate that MobMN243 is capable of nicking the DNA substrate independently of its topology and that the amino acids 200 to 243 modulate substrate specificity but not the nicking activity per se. These findings suggest that these amino acids are involved in positioning the DNA for the nuclease reaction rather than in the nicking mechanism itself.

  12. Transformation of Nickelalactones to Methyl Acrylate: On the Way to a Catalytic Conversion of Carbon Dioxide

    KAUST Repository

    Lee, S. Y. Tina

    2011-08-26

    Mu-nick: The methyl iodide-mediated ring opening of nickelalactones, which can be formed by oxidative coupling of carbon dioxide and ethylene at Ni 0 complexes, induces β-H elimination, producing methyl acrylate in yields of up to 56 %. This reaction is found to be very sensitive to the ligands coordinated to the central nickel atom. © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  13. TeleWound care – providing remote wound assessment and treatment in the home care setting: current status and future directions

    OpenAIRE

    Santamaria, Nick; Kapp,Suzanne

    2013-01-01

    Nick Santamaria,1 Suzanne Kapp2 1University of Melbourne and Melbourne Health, Royal Melbourne Hospital, Melbourne, VIC, Australia; 2Royal District Nursing Service Institute, Melbourne, VIC, Australia Abstract: The use of wound telemedicine systems in the home care environment has been expanding for the last decade. These systems can generally be grouped into two main types: store and forward systems and video conference type systems; additionally, there are also hybrid systems available tha...

  14. TeleWound care – providing remote wound assessment and treatment in the home care setting: current status and future directions

    OpenAIRE

    Santamaria N; Kapp S

    2013-01-01

    Nick Santamaria,1 Suzanne Kapp2 1University of Melbourne and Melbourne Health, Royal Melbourne Hospital, Melbourne, VIC, Australia; 2Royal District Nursing Service Institute, Melbourne, VIC, Australia Abstract: The use of wound telemedicine systems in the home care environment has been expanding for the last decade. These systems can generally be grouped into two main types: store and forward systems and video conference type systems; additionally, there are also hybrid systems available tha...

  15. A Mitochondria-Dependent Pathway Mediates the Apoptosis of GSE-Induced Yeast

    OpenAIRE

    Cao, Sishuo; Xu, Wentao; Zhang, Nan; Wang, Yan; Luo, YunBo; He, Xiaoyun; Huang, Kunlun

    2012-01-01

    Grapefruit seed extract (GSE), which has powerful anti-fungal activity, can induce apoptosis in S. cerevisiae. The yeast cells underwent apoptosis as determined by testing for apoptotic markers of DNA cleavage and typical chromatin condensation by Terminal Deoxynucleotidyl Transferase-mediated dUTP Nick End Labeling (TUNEL) and 4,6'-diaminidino-2-phenylindole (DAPI) staining and electron microscopy. The changes of ΔΨmt (mitochondrial transmembrane potential) and ROS (reactive oxygen species) ...

  16. Palmse viinaköögi manifest

    Index Scriptorium Estoniae

    1996-01-01

    Manifest. Tõlkijate seminari puhul 17. mail 1996 Palmse viinaköögis Juta Bedia, Eric Dickens, Gyözö Fehérvári, Guntars Godinsh, Maima Grinberga, Irja Grönholm, Leif Hannibal, Cornelius Hasselblatt, Pirkko Huurto, Ivo Iliste, Nora Javorskaja, Gisbert Jänicke, Kaisu Lahikainen, Enel Melberg, Vera Ruber, Arnold Tamm, Leili Tamm, Jonatan Tomesh, Lilia Velic'kova, Marianne Vogel. Inglise k.: Estonian Literary Magazine, autumn 1996, nr. 3

  17. The Prado Dam and Reservoir, Riverside and San Bernardino Counties, California

    Science.gov (United States)

    1989-10-31

    County’s Renewed Push for Water Conservation ............. 72 Riverside County Reaction , Late 1940s ........................... 76 Development of...is sloped to the typography to reduce erosion below the concrete-lined section. The emergency spillway had a designed pond elevation of 556 feet, and a...means of pumping water downstream (Nick Richardson, personal communication 1989). 75 4R CL 44- t,, v I. 76 Riverside County Reaction , Late 1940s The

  18. CRITICAL MASS

    Institute of Scientific and Technical Information of China (English)

    DAVID; DAWSON

    2015-01-01

    The Middle Kingdom’s booming Christian community celebrates Christmas教会里的圣诞节并不是购物与狂欢Worldwide,there are two kinds of Christmas.While both are a time of joy,one is relatively secular;its trappings are Christmas trees,gifts,reindeer,and the jolly,red,fat version of Saint Nick(going by圣诞老人,or Christmas Elderly Person in China).

  19. Effects of TEL Confusers on Operator Target Acquisition Performance with SAR Imagery

    Science.gov (United States)

    1998-12-01

    efforts of everyone involved in the project; and Dr. Nick Patton, for providing assistance with several of the mathematical procedures in the document...processing known as the theory of signal detection (TSD) (Gescheider, 1985; Green & Swets, 1966; Macmillan & Creelman , 1991; Wilson, 1992). A TSD...localizations (Hacker & Ratcliff, 1979; Macmillan & Creelman , 1991). The index of bias in a target localization task provides a measure of the operator’s

  20. Combating Terrorism: A Case Study of Nigeria Against Boko Haram Terrorist Group

    Science.gov (United States)

    2014-06-13

    Many scholars on terrorism have pointed to a sense of alienation felt by immigrants , particularly those living in Europe, as a driver of terrorism.39...culture most times are different from those of the immigrants . As a result, they become irritated and begin to feel excluded from the society around...158Nick Chiles , “After Rejecting Nigeria’s Amnesty Offer, Boko Haram Continues to Kill,” Atlanta

  1. Application of DNA Machineries for the Barcode Patterned Detection of Genes or Proteins.

    Science.gov (United States)

    Zhou, Zhixin; Luo, Guofeng; Wulf, Verena; Willner, Itamar

    2018-06-05

    The study introduces an analytical platform for the detection of genes or aptamer-ligand complexes by nucleic acid barcode patterns generated by DNA machineries. The DNA machineries consist of nucleic acid scaffolds that include specific recognition sites for the different genes or aptamer-ligand analytes. The binding of the analytes to the scaffolds initiate, in the presence of the nucleotide mixture, a cyclic polymerization/nicking machinery that yields displaced strands of variable lengths. The electrophoretic separation of the resulting strands provides barcode patterns for the specific detection of the different analytes. Mixtures of DNA machineries that yield, upon sensing of different genes (or aptamer ligands), one-, two-, or three-band barcode patterns are described. The combination of nucleic acid scaffolds acting, in the presence of polymerase/nicking enzyme and nucleotide mixture, as DNA machineries, that generate multiband barcode patterns provide an analytical platform for the detection of an individual gene out of many possible genes. The diversity of genes (or other analytes) that can be analyzed by the DNA machineries and the barcode patterned imaging is given by the Pascal's triangle. As a proof-of-concept, the detection of one of six genes, that is, TP53, Werner syndrome, Tay-Sachs normal gene, BRCA1, Tay-Sachs mutant gene, and cystic fibrosis disorder gene by six two-band barcode patterns is demonstrated. The advantages and limitations of the detection of analytes by polymerase/nicking DNA machineries that yield barcode patterns as imaging readout signals are discussed.

  2. Kinetic mechanism of human DNA ligase I reveals magnesium-dependent changes in the rate-limiting step that compromise ligation efficiency.

    Science.gov (United States)

    Taylor, Mark R; Conrad, John A; Wahl, Daniel; O'Brien, Patrick J

    2011-07-01

    DNA ligase I (LIG1) catalyzes the ligation of single-strand breaks to complete DNA replication and repair. The energy of ATP is used to form a new phosphodiester bond in DNA via a reaction mechanism that involves three distinct chemical steps: enzyme adenylylation, adenylyl transfer to DNA, and nick sealing. We used steady state and pre-steady state kinetics to characterize the minimal mechanism for DNA ligation catalyzed by human LIG1. The ATP dependence of the reaction indicates that LIG1 requires multiple Mg(2+) ions for catalysis and that an essential Mg(2+) ion binds more tightly to ATP than to the enzyme. Further dissection of the magnesium ion dependence of individual reaction steps revealed that the affinity for Mg(2+) changes along the reaction coordinate. At saturating concentrations of ATP and Mg(2+) ions, the three chemical steps occur at similar rates, and the efficiency of ligation is high. However, under conditions of limiting Mg(2+), the nick-sealing step becomes rate-limiting, and the adenylylated DNA intermediate is prematurely released into solution. Subsequent adenylylation of enzyme prevents rebinding to the adenylylated DNA intermediate comprising an Achilles' heel of LIG1. These ligase-generated 5'-adenylylated nicks constitute persistent breaks that are a threat to genomic stability if they are not repaired. The kinetic and thermodynamic framework that we have determined for LIG1 provides a starting point for understanding the mechanism and specificity of mammalian DNA ligases.

  3. Structure-guided Mutational Analysis of the Nucleotidyltransferase Domain of Escherichia coli DNA Ligase (LigA).

    Science.gov (United States)

    Wang, Li Kai; Zhu, Hui; Shuman, Stewart

    2009-03-27

    NAD(+)-dependent DNA ligases (LigA) are ubiquitous in bacteria, where they are essential for growth and present attractive targets for antimicrobial drug discovery. LigA has a distinctive modular structure in which a nucleotidyltransferase catalytic domain is flanked by an upstream NMN-binding module and by downstream OB-fold, zinc finger, helix-hairpin-helix, and BRCT domains. Here we conducted a structure-function analysis of the nucleotidyltransferase domain of Escherichia coli LigA, guided by the crystal structure of the LigA-DNA-adenylate intermediate. We tested the effects of 29 alanine and conservative mutations at 15 amino acids on ligase activity in vitro and in vivo. We thereby identified essential functional groups that coordinate the reactive phosphates (Arg(136)), contact the AMP adenine (Lys(290)), engage the phosphodiester backbone flanking the nick (Arg(218), Arg(308), Arg(97) plus Arg(101)), or stabilize the active domain fold (Arg(171)). Finer analysis of the mutational effects revealed step-specific functions for Arg(136), which is essential for the reaction of LigA with NAD(+) to form the covalent ligase-AMP intermediate (step 1) and for the transfer of AMP to the nick 5'-PO(4) to form the DNA-adenylate intermediate (step 2) but is dispensable for phosphodiester formation at a preadenylylated nick (step 3).

  4. Method for introducing unidirectional nested deletions

    Science.gov (United States)

    Dunn, J.J.; Quesada, M.A.; Randesi, M.

    1999-07-27

    Disclosed is a method for the introduction of unidirectional deletions in a cloned DNA segment. More specifically, the method comprises providing a recombinant DNA construct comprising a DNA segment of interest inserted in a cloning vector. The cloning vector has an f1 endonuclease recognition sequence adjacent to the insertion site of the DNA segment of interest. The recombinant DNA construct is then contacted with the protein pII encoded by gene II of phage f1 thereby generating a single-stranded nick. The nicked DNA is then contacted with E. coli Exonuclease III thereby expanding the single-stranded nick into a single-stranded gap. The single-stranded gapped DNA is then contacted with a single-strand-specific endonuclease thereby producing a linearized DNA molecule containing a double-stranded deletion corresponding in size to the single-stranded gap. The DNA treated in this manner is then incubated with DNA ligase under conditions appropriate for ligation. Also disclosed is a method for producing single-stranded DNA probes. In this embodiment, single-stranded gapped DNA, produced as described above, is contacted with a DNA polymerase in the presence of labeled nucleotides to fill in the gap. This DNA is then linearized by digestion with a restriction enzyme which cuts outside the DNA segment of interest. The product of this digestion is then denatured to produce a labeled single-stranded nucleic acid probe. 1 fig.

  5. AID-induced decrease in topoisomerase 1 induces DNA structural alteration and DNA cleavage for class switch recombination.

    Science.gov (United States)

    Kobayashi, Maki; Aida, Masatoshi; Nagaoka, Hitoshi; Begum, Nasim A; Kitawaki, Yoko; Nakata, Mikiyo; Stanlie, Andre; Doi, Tomomitsu; Kato, Lucia; Okazaki, Il-mi; Shinkura, Reiko; Muramatsu, Masamichi; Kinoshita, Kazuo; Honjo, Tasuku

    2009-12-29

    To initiate class switch recombination (CSR) activation-induced cytidine deaminase (AID) induces staggered nick cleavage in the S region, which lies 5' to each Ig constant region gene and is rich in palindromic sequences. Topoisomerase 1 (Top1) controls the supercoiling of DNA by nicking, rotating, and religating one strand of DNA. Curiously, Top1 reduction or AID overexpression causes the genomic instability. Here, we report that the inactivation of Top1 by its specific inhibitor camptothecin drastically blocked both the S region cleavage and CSR, indicating that Top1 is responsible for the S region cleavage in CSR. Surprisingly, AID expression suppressed Top1 mRNA translation and reduced its protein level. In addition, the decrease in the Top1 protein by RNA-mediated knockdown augmented the AID-dependent S region cleavage, as well as CSR. Furthermore, Top1 reduction altered DNA structure of the Smu region. Taken together, AID-induced Top1 reduction alters S region DNA structure probably to non-B form, on which Top1 can introduce nicks but cannot religate, resulting in S region cleavage.

  6. Unique Helicase Determinants in the Essential Conjugative TraI Factor from Salmonella enterica Serovar Typhimurium Plasmid pCU1

    Energy Technology Data Exchange (ETDEWEB)

    McLaughlin, Krystle J.; Nash, Rebekah P.; Redinbo, Mathew R. (UNC)

    2014-06-16

    The widespread development of multidrug-resistant bacteria is a major health emergency. Conjugative DNA plasmids, which harbor a wide range of antibiotic resistance genes, also encode the protein factors necessary to orchestrate the propagation of plasmid DNA between bacterial cells through conjugative transfer. Successful conjugative DNA transfer depends on key catalytic components to nick one strand of the duplex DNA plasmid and separate the DNA strands while cell-to-cell transfer occurs. The TraI protein from the conjugative Salmonella plasmid pCU1 fulfills these key catalytic roles, as it contains both single-stranded DNA-nicking relaxase and ATP-dependent helicase domains within a single, 1,078-residue polypeptide. In this work, we unraveled the helicase determinants of Salmonella pCU1 TraI through DNA binding, ATPase, and DNA strand separation assays. TraI binds DNA substrates with high affinity in a manner influenced by nucleic acid length and the presence of a DNA hairpin structure adjacent to the nick site. TraI selectively hydrolyzes ATP, and mutations in conserved helicase motifs eliminate ATPase activity. Surprisingly, the absence of a relatively short (144-residue) domain at the extreme C terminus of the protein severely diminishes ATP-dependent strand separation. Collectively, these data define the helicase motifs of the conjugative factor TraI from Salmonella pCU1 and reveal a previously uncharacterized C-terminal functional domain that uncouples ATP hydrolysis from strand separation activity.

  7. Rewetting of a hot metallic wall by liquid spray

    International Nuclear Information System (INIS)

    Castiglia, F.; Giardina, M.; Lombardo, C.

    2005-01-01

    account for the water flow rate, and tested as regards its capability to describe the ENEA experimental results. In the present paper we report the obtained results which seem quite satisfactory. References: [1] G.P. Celata, M. Cumo, C. Lombardo, A. Mariani and L. Saraceno (2003): Experimental result on rewetting of hot surfaces by droplet impingement, '3. European-Japanese Two-Phase Flow Group Meeting', Certosa di Pontignano, 21 - 27 September 2003; [2] F. Castiglia, M. Giardina and E. Oliveri (1996): The heat transfer mechanisms at the quench front, '4. International Symposium on Heat Transfer', 7 - 11 October, Beijing, China. (authors)

  8. Rewetting of a hot metallic wall by liquid spray

    Energy Technology Data Exchange (ETDEWEB)

    Castiglia, F.; Giardina, M.; Lombardo, C. [University of Palermo, Department of Nuclear Engineering, V.le delle Scienze, 90128 Palermo (Italy)

    2005-07-01

    suitably modified in order to account for the water flow rate, and tested as regards its capability to describe the ENEA experimental results. In the present paper we report the obtained results which seem quite satisfactory. References: [1] G.P. Celata, M. Cumo, C. Lombardo, A. Mariani and L. Saraceno (2003): Experimental result on rewetting of hot surfaces by droplet impingement, '3. European-Japanese Two-Phase Flow Group Meeting', Certosa di Pontignano, 21 - 27 September 2003; [2] F. Castiglia, M. Giardina and E. Oliveri (1996): The heat transfer mechanisms at the quench front, '4. International Symposium on Heat Transfer', 7 - 11 October, Beijing, China. (authors00.

  9. Plasmid DNA damage caused by stibine and trimethylstibine

    International Nuclear Information System (INIS)

    Andrewes, Paul; Kitchin, Kirk T.; Wallace, Kathleen

    2004-01-01

    Antimony is classified as 'possibly carcinogenic to humans' and there is also sufficient evidence for antimony carcinogenicity in experimental animals. Stibine is a volatile inorganic antimony compound to which humans can be exposed in occupational settings (e.g., lead-acid battery charging). Because it is highly toxic, stibine is considered a significant health risk; however, its genotoxicity has received little attention. For the work reported here, stibine was generated by sodium borohydride reduction of potassium antimony tartrate. Trimethylstibine is a volatile organometallic antimony compound found commonly in landfill and sewage fermentation gases at concentrations ranging between 0.1 and 100 μg/m 3 . Trimethylstibine is generally considered to pose little environmental or health risk. In the work reported here, trimethylstibine was generated by reduction of trimethylantimony dichloride using either sodium borohydride or the thiol compounds, dithioerythritol (DTE), L-cysteine, and glutathione. Here we report the evaluation of the in vitro genotoxicities of five antimony compounds--potassium antimony tartrate, stibine, potassium hexahydroxyantimonate, trimethylantimony dichloride, and trimethylstibine--using a plasmid DNA-nicking assay. Of these five antimony compounds, only stibine and trimethylstibine were genotoxic (significant nicking to pBR 322 plasmid DNA). We found stibine and trimethylstibine to be about equipotent with trimethylarsine using this plasmid DNA-nicking assay. Reaction of trimethylantimony dichloride with either glutathione or L-cysteine to produce DNA-damaging trimethylstibine was observed with a trimethylantimony dichloride concentration as low as 50 μM and L-cysteine or glutathione concentrations as low as 500 and 200 μM, respectively, for a 24 h incubation

  10. The Role of XPG in Processing (CAGn/(CTGn DNA Hairpins

    Directory of Open Access Journals (Sweden)

    Hou Caixia

    2011-03-01

    Full Text Available Abstract Background During DNA replication or repair, disease-associated (CAGn/(CTGn expansion can result from formation of hairpin structures in the repeat tract of the newly synthesized or nicked DNA strand. Recent studies identified a nick-directed (CAGn/(CTGn hairpin repair (HPR system that removes (CAGn/(CTGn hairpins from human cells via endonucleolytic incisions. Because the process is highly similar to the mechanism by which XPG and XPF endonucleases remove bulky DNA lesions during nucleotide excision repair, we assessed the potential role of XPG in conducting (CAGn/(CTGn HPR. Results To determine if the XPG endonuclease is involved in (CAGn/(CTGn hairpin removal, two XPG-deficient cell lines (GM16024 and AG08802 were examined for their ability to process (CAGn/(CTGn hairpins in vitro. We demonstrated that the GM16024 cell line processes all hairpin substrates as efficiently as HeLa cells, and that the AG08802 cell line is partially defective in HPR. Analysis of repair intermediates revealed that nuclear extracts from both XPG-deficient lines remove CAG/CTG hairpins via incisions, but the incision products are distinct from those generated in HeLa extracts. We also show that purified recombinant XPG protein greatly stimulates HPR in XPG-deficient extracts by promoting an incision 5' to the hairpin. Conclusions Our results strongly suggest that 1 human cells possess multiple pathways to remove (CAGn/(CTGn hairpins located in newly synthesized (or nicked DNA strand; and 2 XPG, although not essential for (CAGn/(CTGn hairpin removal, stimulates HPR by facilitating a 5' incision to the hairpin. This study reveals a novel role for XPG in genome-maintenance and implicates XPG in diseases caused by trinucleotide repeat expansion.

  11. Activation of Saccharomyces cerevisiae Mlh1-Pms1 Endonuclease in a Reconstituted Mismatch Repair System*

    Science.gov (United States)

    Smith, Catherine E.; Bowen, Nikki; Graham, William J.; Goellner, Eva M.; Srivatsan, Anjana; Kolodner, Richard D.

    2015-01-01

    Previous studies reported the reconstitution of an Mlh1-Pms1-independent 5′ nick-directed mismatch repair (MMR) reaction using Saccharomyces cerevisiae proteins. Here we describe the reconstitution of a mispair-dependent Mlh1-Pms1 endonuclease activation reaction requiring Msh2-Msh6 (or Msh2-Msh3), proliferating cell nuclear antigen (PCNA), and replication factor C (RFC) and a reconstituted Mlh1-Pms1-dependent 3′ nick-directed MMR reaction requiring Msh2-Msh6 (or Msh2-Msh3), exonuclease 1 (Exo1), replication protein A (RPA), RFC, PCNA, and DNA polymerase δ. Both reactions required Mg2+ and Mn2+ for optimal activity. The MMR reaction also required two reaction stages in which the first stage required incubation of Mlh1-Pms1 with substrate DNA, with or without Msh2-Msh6 (or Msh2-Msh3), PCNA, and RFC but did not require nicking of the substrate, followed by a second stage in which other proteins were added. Analysis of different mutant proteins demonstrated that both reactions required a functional Mlh1-Pms1 endonuclease active site, as well as mispair recognition and Mlh1-Pms1 recruitment by Msh2-Msh6 but not sliding clamp formation. Mutant Mlh1-Pms1 and PCNA proteins that were defective for Exo1-independent but not Exo1-dependent MMR in vivo were partially defective in the Mlh1-Pms1 endonuclease and MMR reactions, suggesting that both reactions reflect the activation of Mlh1-Pms1 seen in Exo1-independent MMR in vivo. The availability of this reconstituted MMR reaction should now make it possible to better study both Exo1-independent and Exo1-dependent MMR. PMID:26170454

  12. Activation of Saccharomyces cerevisiae Mlh1-Pms1 Endonuclease in a Reconstituted Mismatch Repair System.

    Science.gov (United States)

    Smith, Catherine E; Bowen, Nikki; Graham, William J; Goellner, Eva M; Srivatsan, Anjana; Kolodner, Richard D

    2015-08-28

    Previous studies reported the reconstitution of an Mlh1-Pms1-independent 5' nick-directed mismatch repair (MMR) reaction using Saccharomyces cerevisiae proteins. Here we describe the reconstitution of a mispair-dependent Mlh1-Pms1 endonuclease activation reaction requiring Msh2-Msh6 (or Msh2-Msh3), proliferating cell nuclear antigen (PCNA), and replication factor C (RFC) and a reconstituted Mlh1-Pms1-dependent 3' nick-directed MMR reaction requiring Msh2-Msh6 (or Msh2-Msh3), exonuclease 1 (Exo1), replication protein A (RPA), RFC, PCNA, and DNA polymerase δ. Both reactions required Mg(2+) and Mn(2+) for optimal activity. The MMR reaction also required two reaction stages in which the first stage required incubation of Mlh1-Pms1 with substrate DNA, with or without Msh2-Msh6 (or Msh2-Msh3), PCNA, and RFC but did not require nicking of the substrate, followed by a second stage in which other proteins were added. Analysis of different mutant proteins demonstrated that both reactions required a functional Mlh1-Pms1 endonuclease active site, as well as mispair recognition and Mlh1-Pms1 recruitment by Msh2-Msh6 but not sliding clamp formation. Mutant Mlh1-Pms1 and PCNA proteins that were defective for Exo1-independent but not Exo1-dependent MMR in vivo were partially defective in the Mlh1-Pms1 endonuclease and MMR reactions, suggesting that both reactions reflect the activation of Mlh1-Pms1 seen in Exo1-independent MMR in vivo. The availability of this reconstituted MMR reaction should now make it possible to better study both Exo1-independent and Exo1-dependent MMR. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

  13. Detailed Measurements of Rayleigh-Taylor Mixing at Large and Small Atwood Numbers

    International Nuclear Information System (INIS)

    Malcolm, J.; Andrews, Ph.D.

    2004-01-01

    This project has two major tasks: Task 1. The construction of a new air/helium facility to collect detailed measurements of Rayleigh-Taylor (RT) mixing at high Atwood number, and the distribution of these data to LLNL, LANL, and Alliance members for code validation and design purposes. Task 2. The collection of initial condition data from the new Air/Helium facility, for use with validation of RT simulation codes at LLNL and LANL. Also, studies of multi-layer mixing with the existing water channel facility. Over the last twelve (12) months there has been excellent progress, detailed in this report, with both tasks. As of December 10, 2004, the air/helium facility is now complete and extensive testing and validation of diagnostics has been performed. Currently experiments with air/helium up to Atwood numbers of 0.25 (the maximum is 0.75, but the highest Reynolds numbers are at 0.25) are being performed. The progress matches the project plan, as does the budget, and we expect this to continue for 2005. With interest expressed from LLNL we have continued with initial condition studies using the water channel. This work has also progressed well, with one of the graduate Research Assistants (Mr. Nick Mueschke) visiting LLNL the past two summers to work with Dr. O. Schilling. Several journal papers are in preparation that describe the work. Two MSc.'s have been completed (Mr. Nick Mueschke, and Mr. Wayne Kraft, 12/1/03). Nick and Wayne are both pursuing Ph.D.s' funded by this DOE Alliances project. Presently three (3) Ph.D. graduate Research Assistants are supported on the project, and two (2) undergraduate Research Assistants. During the year two (2) journal papers and two (2) conference papers have been published, ten (10) presentations made at conferences, and three (3) invited presentations

  14. Characterization of the adenoassociated virus Rep protein complex formed on the viral origin of DNA replication

    International Nuclear Information System (INIS)

    Li Zengi; Brister, J. Rodney; Im, Dong-Soo; Muzyczka, Nicholas

    2003-01-01

    Interaction between the adenoassociated virus (AAV) replication proteins, Rep68 and 78, and the viral terminal repeats (TRs) is mediated by a DNA sequence termed the Rep-binding element (RBE). This element is necessary for Rep-mediated unwinding of duplex DNA substrates, directs Rep catalyzed cleavage of the AAV origin of DNA replication, and is required for viral transcription and proviral integration. Six discrete Rep complexes with the AAV TR substrates have been observed in vitro, and cross-linking studies suggest these complexes contain one to six molecules of Rep. However, the functional relationship between Rep oligomerization and biochemical activity is unclear. Here we have characterized Rep complexes that form on the AAV TR. Both Rep68 and Rep78 appear to form the same six complexes with the AAV TR, and ATP seems to stimulate formation of specific, higher order complexes. When the sizes of these Rep complexes were estimated on native polyacrylamide gels, the four slower migrating complexes were larger than predicted by an amount equivalent to one or two TRs. To resolve this discrepancy, the molar ratio of protein and DNA was calculated for the three largest complexes. Data from these experiments indicated that the larger complexes included multiple TRs in addition to multiple Rep molecules and that the Rep-to-TR ratio was approximately 2. The two largest complexes were also associated with increased Rep-mediated, origin cleavage activity. Finally, we characterized a second, Rep-mediated cleavage event that occurs adjacent to the normal nicking site, but on the opposite strand. This second site nicking event effectively results in double-stranded DNA cleavage at the normal nicking site

  15. Transumanesimo, enhancement ed evoluzione. Riflessioni preliminari per una ridefinizione critica

    Directory of Open Access Journals (Sweden)

    LO SAPIO, LUCA

    2016-12-01

    Full Text Available Transhumanism, enhancement and evolution. Preliminary reflections for a critical redefinition This paper aims at focusing some possible uses of evolutionary theory in transhumanist authors. In particular it is analysed the evolutionary heuristics discussed by Nick Bostrom and Anders Sandberg, the proposal of a moral bioenhancement in Julian Savulescu and Ingmar Persson and the idea itself of a self-direct evolution based on the assumption of a self-guided selection of men’s features. These ideas are discussed to bring out the critical elements which are present in some uses of a presumably Darwinian framework in transhumanist approach.

  16. The UKIRT Upgrades Programme

    Science.gov (United States)

    Adamson, Andy; Davies, John; Robson, Ian

    Tim Hawarden presented this paper to the 30th anniversary workshop, just a month before his untimely death. The editors have done their best to convert his talk into this paper, and gratefully acknowledge the assistance of Nick Rees (a member of the Upgrades team, now at Diamond Light Source). Tim's discussion concerned the UKIRT Upgrades Project, which ran through the 1990s and transformed the telescope and made it truly competitive on the world stage for operation into the twenty-first century. The reference list at the end of the paper is comprehensive; some of these are referred to in the paper itself and some are included for completeness only.

  17. Loglines. November-December 2012

    Science.gov (United States)

    2012-12-01

    other experience and skills that are a good fit for a manager,” she said. Clay Henson, a former Army infantry- man, was one of the first service...new employees need to know that the agency provides? DLA supports warfighters. DLA provides clothing, medicine, meals , fuel and replacement parts for...first-strike ration is smaller and lighter than a meal , ready-to-eat. It is used by the U.S. Michael Tuttle and Nick Sistrun are writers for DLA

  18. Electric power systems

    CERN Document Server

    Weedy, B M; Jenkins, N; Ekanayake, J B; Strbac, G

    2012-01-01

    The definitive textbook for Power Systems students, providing a grounding in essential power system theory while also focusing on practical power engineering applications. Electric Power Systems has been an essential book in power systems engineering for over thirty years. Bringing the content firmly up-to-date whilst still retaining the flavour of Weedy's extremely popular original, this Fifth Edition has been revised by experts Nick Jenkins, Janaka Ekanayake and Goran Strbac. This wide-ranging text still covers all of the fundamental power systems subjects but is now e

  19. First-generation physical map of the Culicoides variipennis (Diptera: Ceratopogonidae) genome.

    Science.gov (United States)

    Nunamaker, R A; Brown, S E; McHolland, L E; Tabachnick, W J; Knudson, D L

    1999-11-01

    Recombinant cosmids labeled with biotin-11-dUTP or digoxigenin by nick translation were used as in situ hybridization probes to metaphase chromosomes of Culicoides variipennis (Coquillett). Paired fluorescent signals were detected on each arm of sister chromatids and were ordered along the 3 chromosomes. Thirty-three unique probes were mapped to the 3 chromosomes of C. variipennis (2n = 6): 7 to chromosome 1, 20 to chromosome 2, and 6 to chromosome 3. This work represents the first stage in generating a physical map of the genome of C. variipennis.

  20. Leishmania diagnostic and identification py using 32P labelled DNA probes

    International Nuclear Information System (INIS)

    Andrade, Antero Silva Ribeiro de; Melo, Maria Norma de

    1999-10-01

    P 32 labelled DNA probes are valious instruments for the parasitic diseases by using hybridization reaction. In this paper we describe the methodology and present the foundations for the radioactive probes production, based on the kinetoplast DNA (kDNA), for the Leishmania diagnostic an identification. We also describe the kDNA purification protocol from Leishmania reference cepa, the process of P 32 labelling of the kDNA by using the nick translation method, gathering, sample preparation and treatment, the optimum conditions for the hybridization reaction and the procedures for the autoradiography

  1. Work on CMS Muon Detector (CSCs) during Long Shutdown 1

    CERN Multimedia

    Lapka, Marzena

    2015-01-01

    These U.S. physicists made crucial contributions in preparing new electronics in the CMS effort to refurbish the first layer of the muon detector endcap. This effort recently passed a major review, suggesting the project is on track to be part of the experiment when detector operations resumes in early 2015. From left to right: Wells Wulsin (Oklahoma State), Nick Amin (Texas A&M), Indara Suarez (Texas A&M), Shalhout Shalhout (UC Davis), Joe Haley (Northeastern), Michael Gardner (UC Davis), and Justin Pilot (UC Davis). Inset: Joe Golf (left) and Manuel Franco Sevilla, both of UC Santa Barbara.

  2. Introduction to the Volume's Two Thematic Sections

    Directory of Open Access Journals (Sweden)

    Christopher Chase-Dunn

    2015-08-01

    Full Text Available The first batch contains an article by Political Scientist Daniel Whitcncck on epistemic communities and global leadership and a special thematic section focussing around the ideas ofW. Warren Wagar regarding the future of the world-system and global politics. The second batch of papers in Volume 2 has been edited by P. Nick Kardulias of the Department of Anthropology and Sociology at Kenyon College. This special thematic section focusses on anthropological and archaeological approaches to the study of world systems.

  3. Study of radionuclide 90Sr-90Y on cell proliferation and apoptosis in benign prostatic hyperplasia

    International Nuclear Information System (INIS)

    Zhang Tong; Wei Wei; Zou Benjie; Liu Fang; Xu Zhishun

    2003-01-01

    Objective: To investigate the effect of 90 Sr- 90 Yon cell proliferation and apoptosis in benign prostatic hyperplasia. Methods: The apoptosis and expression of Ki-67 in benign prostatic hyperplasia (BPH) before and after irradiation 90 Sr- 90 Y were detected by transferase-mediated dUTP-biotin nick end labeling (TUNEL) method and immunohistochemical technique, respectively. Results: The proliferation index (PI) of BPH after 90 Sr- 90 Y irradiation was much lower than that before irradiation, but there was no significant change in apoptosis index (AI). Conclusion: Irradiation with 90 Sr- 90 Y could restrain cell proliferation of BPH, but could not induce apoptosis

  4. History of endeavours to mitigate the rockburst risk in South African mines

    CSIR Research Space (South Africa)

    Durrheim, RJ

    2012-04-01

    Full Text Available for periods of a few months to a few years. ?Golden decade? of mine seismology at BPI Geophysics (1969-1979) From 1969 to 1979 the BPI Geophysics team of Art McGarr, Steve Spottiswoode, Rod Green and Nick Gay contributed significantly to the emerging... discipline of mine seismology (e.g. McGarr 1971; Spottiswoode and McGarr 1975; McGarr et al. 1975; Gay and Ortlepp 1979 and McGarr et al. 1981). Much of this work was carried out at ERPM. The magnitudes of the stresses driving violent failure...

  5. Talking about Life

    Science.gov (United States)

    Impey, Chris

    2010-08-01

    Preface; Part I. Introduction Timothy Ferris, Iris Fry, Steven Dick, Ann Druyan, Pinky Nelson, Neil Tyson, Steve Benner and William Bains; Part II. Earth Roger Buick, Lynn Rothschild, John Baross, Joe Kirschvink, Andrew Knoll, Simon Conway Morris, Roger Hanlon and Lori Marino; Part III. Solar System Chris McKay, David Grinspoon, Jonathan Lunine, Carolyn Porco, Laurie Leshin, Guy Consolmagno and Peter Smith; Part IV. Exoplanets Alan Boss, Geoff Marcy, Debra Fischer, Sara Seager, David Charbonneau and Vikki Meadows; Part V. Frontiers Jill Tarter, Seth Shostak, Ray Kurzweil, Nick Bostrom, Paul Davies, Martin Rees, Ben Bova and Jennifer Michael Hecht; Reading list; Glossary; Index.

  6. The Mirror in the Ground

    DEFF Research Database (Denmark)

    Shepherd, Nick

    An important and original contribution to the study of the archive, The Mirror in the Ground approaches the discipline of archaeology in South Africa from the perspective of an interest in visualities. Author Nick Shepherd argues that it makes sense to talk about an archaeological aesthetics...... at the University of Cape Town, where he convenes a graduate programme on Public Culture and Heritage. The Mirror in the Ground is the first volume in the relaunched Series in Visual Histories, produced by the Centre for Curating the Archive (CCA) at the University of Cape Town....

  7. Binding of anthracene to cellular macromolecules in the presence of light. [DNA, HSA

    Energy Technology Data Exchange (ETDEWEB)

    Sinha, B K; Chignell, C F [National Inst. of Environmental Health Sciences, Research Triangle Park, NC (USA)

    1983-01-01

    Ultraviolet radiation (> 295 nm) induced covalent binding of anthracene to DNA which increased with time and was not affected by oxygen. Irradiation in the presence of anthracene induced nicking of Col E/sub 1/ circular DNA and decreased the thermal denaturation temperature of calf thymus DNA. These effects were oxygen dependent, and were decreased by GMP. Irradiation of anthracene and human serum albumin resulted in covalent binding of the hydrocarbon to the protein accompanied by crosslinking of the protein. Protein crosslinking decreased under anaerobic conditions. Irradiation of anthracene bound to liposomes induced lipid peroxidation which was not affected by superoxide dismutase or catalase.

  8. Delayed manifestation of bilateral scleral thinning after I-BRITE® procedure and review of literature for cosmetic eye-whitening procedures

    OpenAIRE

    Fenzl, Carlton; McCaughey,Michael; Santiago-Caban,Luis; Kramer,Gregory; Mamalis,Nick; Majid Moshirfar,Majid

    2015-01-01

    Majid Moshirfar,1 Michael V McCaughey,2 Carlton R Fenzl,3 Luis Santiago-Caban,4 Gregory D Kramer,3 Nick Mamalis31Department of Ophthalmology, Francis I Proctor Foundation, University of California San Francisco, San Francisco, CA, USA; 2University of New Mexico School of Medicine, Albuquerque, NM, USA; 3John A Moran Eye Center, University of Utah, Salt Lake City, UT, USA; 4Department of Ophthalmology, University of Puerto Rico School of Medicine, San Juan, PR, USAPurpose: To report a case of ...

  9. The effects of ascorbic acid on diphtheria toxin and intoxicated hela cells

    International Nuclear Information System (INIS)

    Clark, C.E.; Smith, T.J.

    1976-01-01

    Ascorbic acid (vitamin C) prevented diphtheria toxin from inhibiting the incorporation of [U- 14 C]-alanine into trichloroacetic acid precipitable material in HeLa cells. Ascorbic acid did not exhibit an effect on the adenosine diphosphate-ribosylation of amino acyl transferase 2 nor did it separate fragment A from fragment B in ''nicked'' toxin. A non-specific reducing agent, para-methylaminophenol sulfate, exhibited an effect of HeLa cells very similar to the results of ascorbic acid. Citric acid, a tricarboxylic acid, had no effect on HeLa cells. (auth.)

  10. Enzymatic quantification of strand breaks of DNA induced by vacuum-UV radiation

    International Nuclear Information System (INIS)

    Ito, Takashi

    1986-01-01

    Hind3 digested plasmid DNA dried on an aluminum plate was irradiated by vacuum-UV at 160 and 195 nm using a synchrotron irradiation system. A change induced in the DNA, presumably a single strand break, was quantified by the aid of the strand break-derived stimulation of poly(ADP-ribose) synthetase activity. The end group of strand breaks so induced was recognized by the enzyme as effectively as that by DNase 1 treatment, suggesting a nicking as the major lesion inflicted on the DNA. The fluence (UV) dependent stimulation of poly(ADP-ribose) synthetase activity was much higher upon 160 nm irradiation than upon 195 nm irradiation. (Auth.)

  11. Human exonuclease 1 (EXO1) activity characterization and its function on FLAP structures

    DEFF Research Database (Denmark)

    Keijzers, Guido; Bohr, Vilhelm A; Juel Rasmussen, Lene

    2015-01-01

    structures, we determined factors essential for the thermodynamic stability of EXO1. We show that enzymatic activity and stability of EXO1 on DNA is modulated by temperature. By characterization of EXO1 flap activity using various DNA flap substrates, we show that EXO1 has a strong capacity for degrading...... double stranded DNA and has a modest endonuclease or 5' flap activity. Furthermore, we report novel mechanistic insights into the processing of flap structures, showing that EXO1 preferentially cleaves one nucleotide inwards in a double stranded region of a forked and nicked DNA flap substrates...

  12. Laser Cutting of Thin Nickel Bellows

    Science.gov (United States)

    Butler, C. L.

    1986-01-01

    Laser cutting technique produces narrow, precise, fast, and repeatable cuts in thin nickel-allow bellows material. Laser cutting operation uses intense focused beam to melt material and assisting gas to force melted material through part thickness, creating void. When part rotated or moved longitudinally, melting and material removal continuous and creates narrow, fast, precise, and repeatable cut. Technique used to produce cuts of specified depths less than material thickness. Avoids distortion, dents, and nicks produced in delicate materials during lathe trimming operations, which require high cutting-tool pressure and holding-fixture forces.

  13. Maculosin, a host-specific phytotoxin for spotted knapweed from Alternaria alternata

    Science.gov (United States)

    Stierle, Andrea C.; Cardellina, John H.; Strobel, Gary A.

    1988-01-01

    Several diketopiperazines have been isolated from liquid cultures of Alternaria alternata, the causal agent of black leaf blight of spotted knapweed, Centaurea maculosa Lam. One of these compounds, maculosin [the diketopiperazine cyclo(-L-Pro-L-Tyr-)], was active in the nicked-leaf bioassay at 10-5 M; synthetic maculosin possessed chemical and biological activities identical to those of the natural product. Other diketopiperazines isolated from the fungus possessed either less activity or none at all. In tests against 19 plant species, maculosin was phytotoxic only to spotted knapweed. Thus maculosin is a host-specific phytotoxin from a weed pathogen. PMID:16593989

  14. Statistical studies for SNP association in acute coronary syndrome ex vivo use of agonists and nanoparticles

    CSIR Research Space (South Africa)

    Das Roy, P

    2014-06-01

    Full Text Available for Scientific and Industrial Research, Pretoria Background information Acute Coronary Syndrome (ACS), nick named as heart attack, is becoming one of the more frequent causes of death in today’s fast paced stressed out life, 4.8% of total death around... it occurs in order to prevent further blood loss during bleeding. However, sometime this platelet aggregation within the blood vessels cause the blocking of blood flow to the heart and hence cause what we know as heart attack, strokes, and peripheral...

  15. Renal cell apoptosis in human lupus nephritis: a histological study

    DEFF Research Database (Denmark)

    Faurschou, M; Penkowa, Milena; Andersen, C B

    2009-01-01

    Nuclear autoantigens from apoptotic cells are believed to drive the immunological response in systemic lupus erythematosus (SLE). Conflicting data exist as to the possible renal origin of apoptotic cells in SLE patients with nephritis. We assessed the level of renal cell apoptosis in kidney...... biopsies from 35 patients with lupus nephritis by means of terminal deoxynucleotidyl-transferase (TdT)-mediated deoxyuridine triphosphate (dUTP)-digoxigenin nick end labeling (TUNEL). Five samples of normal kidney tissue served as control specimens. We did not observe apoptotic glomerular cells in any...... cells constitute a quantitatively important source of auto-antibody-inducing nuclear auto-antigens in human lupus nephritis....

  16. Good Thoughts to Keep in Mind

    Institute of Scientific and Technical Information of China (English)

    刘保行

    2005-01-01

    I magine life as a game in which you are juggling some five balls in the air.You name them:Work,Family,Health,Friends,Spirit.And you are keeping all of these in the air.You will soon understand that work is a rubber ball.lf you drop it,it will bounce back.But the other four balls-family,health,friends and spirit are made of glass.lf you drop one of these,they will be irrevocably scuffed,marked,nicked,damaged or even shattered .They will never be the same.You must understand that and strive for balance inyour life.

  17. Apoptosis of gut-associated lymphoid tissue in rainbow trout Oncorhynchus mykiss after incubation with Candida albicans and bacterial lipopolysaccharide.

    Science.gov (United States)

    Passantino, L; Ostillio, A; Cianciotta, A; Russo, C; Carrassi, M; Patruno, R; Dhaskali, L; Passantino, G F; Passantino, A

    2011-06-01

    Until now a few studies have been carried out on the gut lymphoid system in fish despite its protective role in the host. Here, we have evaluated the effects of Candida albicans (Ca) and lipopolysaccaride (LPS) on the pyloric and terminal segments of gut in the rainbow trout Oncorhynchus mykiss. In particular, data show that both Ca and LPS are able to cause apoptosis of intestinal lymphoid cells as detected by the terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) procedure. These findings suggest a further modality of gut response in fish to environmental antigens.

  18. Artificial Intelligence

    CERN Document Server

    Warwick, Kevin

    2011-01-01

    if AI is outside your field, or you know something of the subject and would like to know more then Artificial Intelligence: The Basics is a brilliant primer.' - Nick Smith, Engineering and Technology Magazine November 2011 Artificial Intelligence: The Basics is a concise and cutting-edge introduction to the fast moving world of AI. The author Kevin Warwick, a pioneer in the field, examines issues of what it means to be man or machine and looks at advances in robotics which have blurred the boundaries. Topics covered include: how intelligence can be defined whether machines can 'think' sensory

  19. Inhibition of histone deacetylases prevents cytokine-induced toxicity in beta cells

    DEFF Research Database (Denmark)

    Larsen, L; Tonnesen, M; Ronn, S G

    2007-01-01

    B (NFkappaB) is a critical signalling molecule in inflammation and is required for expression of the gene encoding inducible NO synthase (iNOS) and of pro-apoptotic genes. NFkappaB has recently been shown to associate with chromatin-modifying enzymes histone acetyltransferases and histone...... by immunoblotting and by immunoblotting combined with electrophoretic mobility shift assay, respectively. Viability was analysed by 3-(4,5-dimethyldiazol-2-yl)-2,5-diphenyl-tetrazolium bromide and apoptosis by terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) assay and histone...

  20. Erratum to: The blocking effect in associative learning involves learned biases in rapid attentional capture.

    Science.gov (United States)

    2018-04-01

    Luque, D., Vadillo, M, A., Gutiérrez-Cobo, M, J., Le Pelley, M, E. (2018). The blocking effect in associative learning involves learned biases in rapid attentional capture. Quarterly Journal of Experimental Psychology, 71, 522-544. doi: 10.1080/17470218.2016.1262435. The above article is part of the Special Issue on Associative Learning (in honour of Nick Mackintosh) and was inadvertently published in the February 2018 issue of Quarterly Journal of Experimental Psychology. After publication of the Special Issue, an online collection on Associative Learning will be created on SAGE Journals and this paper will be included in that collection. The Publisher apologises for this error.

  1. Effect of ionizing radiation on apoptosis in the cortex of mouse lymph node

    International Nuclear Information System (INIS)

    Chen Dong; Liu Jiamei; Liu Shuzheng

    1999-01-01

    Objective: To study the alteration of apoptosis in the cortex of mouse lymph node following whole body X-irradiation. Methods: The method of TdT-mediated dUTP nick end labelling (TUNEL) was used to detect apoptosis the cortex of mouse lymph node. Results: The sensitivity to high and low dose ionizing radiation was distinct in different area of the cortex. Conclusion: The decrease of apoptotic cells in the inter nodular and deep cortex indicate that low dose radiation may suppress the apoptosis of T lymphocytes and play a role in immune regulation

  2. From Affective Shareholding to OUR Walmart: Organizing Labor in a Post-Union World

    Directory of Open Access Journals (Sweden)

    Christine Labuski

    2015-12-01

    Full Text Available From Affective Shareholding to OUR Walmart: Organizing Labor in a Post-Union World, by Christine Labuski and Nick Copeland. This essay examines OUR Walmart (Organization United for Respect at Walmart, a group of current and former Walmart employees who are fighting for improved working conditions within the company. We situate OUR Walmart's emergence within the breakdown of management strategies that substitute affective and symbolic inclusion for material benefits. We also show how OUR Walmart shames the company over social media by contrasting workers’ precarious lives with the extreme wealth of the Walton family, and redefines respect to include collective and material rights.

  3. Editorial - Special Issue on the Ninth International Conference on Aeolian Research - ICAR IX (Coastal Dune Processes and Aeolian Transport)

    Science.gov (United States)

    da Silva, Graziela Miot

    2018-04-01

    This special issue combines some of the papers related to coastal dune processes and aeolian sediment transport that were presented at the Ninth International Conference on Aeolian Research - ICAR IX. The conference was held between 4 and 8 of July 2016 in Mildura, Australia, organized by the International Society for Aeolian Research (ISAR) and convened by Adrian Chappell (Cardiff University), Craig Strong (Australian National University), Stephen Cattle (University of Sydney), Patrick Hesp (Flinders University), John Leys (New South Wales Office of Environment and Heritage), Lynda Petherick (University of Wellington) and Nick Webb (USDA-ARS Jornada Experimental Range).

  4. Worlds within Worlds: Audiences, Jargon, and North American Comics Discourse

    Directory of Open Access Journals (Sweden)

    Craig Fischer

    2010-09-01

    Full Text Available Let me begin by describing four recent acts of comics criticism, all connected to each other like dominoes falling: The first act: In 2007, the University Press of Mississippi published an English-language translation of Thierry Groensteen’s The System of Comics (Système de la bande dessinée, 1999. In their foreword to System, translators Bart Beaty and Nick Nguyen anticipate obstacles to the American reception of Groensteen’s ideas, arguing that American scholars are unfamiliar with the “do...

  5. Areeni aasta albumid 2004

    Index Scriptorium Estoniae

    2004-01-01

    Heliplaatidest: Franz Ferdinand "Franz Ferdinand", Toe Tag "Legendaarne", The Libertines "The Libertines ", Morrissey "You Are The Qarry", The Streets "A Grand Don't Come For Free", Kalm "Generalissimus Kalm", Brian Wilson "Smile", Madvillain "Madvillainy", Scissor Sisters "Scissor Sisters", Gary Wilson "Mary Had A Brown Hair", Animal Collective "Sung Tongs", Keane "Hopes And Fears", Todd Rundgren "Liars", Nick Cave & The Bad Seeds "Abbatoir Blues/The Lyre of Orpheus", Mylo "Destroy Rock'n'Roll", Devendra Banhart "Rejoicing In The Hands", Stefani "Love Angel Music Baby", Björk "Medulla", Moodymann "Black Mahogani"

  6. Study Drugs

    OpenAIRE

    Lam, Stephanie Phuong; Roosta, Natalie; Nielsen, Mikkel Fuhr; Meyer, Maria Holmgaard; Friis, Katrine Birk

    2016-01-01

    In recent years, students around the world, started to use preparations as Ritalin and Modafinil,also known as study drugs, to improve their cognitive abilities1. It is a common use among thestudents in United States of America, but it is a new tendency in Denmark. Our main focus is tolocate whether study drugs needs to be legalized in Denmark or not. To investigate this ourstarting point is to understand central ethical arguments in the debate. We have chosen twoarguments from Nick Bostrom a...

  7. Concentrating and labeling genomic DNA in a nanofluidic array

    DEFF Research Database (Denmark)

    Marie, Rodolphe; Pedersen, Jonas Nyvold; Mir, Kalim U.

    2018-01-01

    , however, hinder the polymerase activity. We demonstrate a device and a protocol for the enzymatic labeling of genomic DNA arranged in a dense array of single molecules without attaching the enzyme or the DNA to a surface. DNA molecules accumulate in a dense array of pits embedded within a nanoslit due...... to entropic trapping. We then perform ϕ29 polymerase extension from single-strand nicks created on the trapped molecules to incorporate fluorescent nucleotides into the DNA. The array of entropic traps can be loaded with λ-DNA molecules to more than 90% of capacity at a flow rate of 10 pL min-1. The final...

  8. Probability and Statistics: 5 Questions

    DEFF Research Database (Denmark)

    Probability and Statistics: 5 Questions is a collection of short interviews based on 5 questions presented to some of the most influential and prominent scholars in probability and statistics. We hear their views on the fields, aims, scopes, the future direction of research and how their work fits...... in these respects. Interviews with Nick Bingham, Luc Bovens, Terrence L. Fine, Haim Gaifman, Donald Gillies, James Hawthorne, Carl Hoefer, James M. Joyce, Joseph B. Kadane Isaac Levi, D.H. Mellor, Patrick Suppes, Jan von Plato, Carl Wagner, Sandy Zabell...

  9. Control design profits heliostat techno-economics

    CSIR Research Space (South Africa)

    Rubin, N

    2012-05-01

    Full Text Available , and the economic impacts associated with reduced spillage at the receiver, that accrue through improved analysis, modelling and design techniques. 2. Background 2.1. Previous Results It is testimony to the robustness of feedback control that a stabilizing... Heliostat Techno-Economics Nick Rubin1, Thomas Roos2, Fintan Wilson2, Jason de Villiers2 and Strinivasan Perumal2 1 Systems Engineer ? CSIR, P O Box 395, Pretoria, 0001, South Africa, Phone: 0128413383, Fax:0128413179, NRubin@csir.co.za 2 THRoos...

  10. Metazoos parásitos de la mojarrilla Stellifer minor (Tschudi (Osteichthyes, Sciaenidae capturados por pesquería artesanal en Chorrillos, Lima, Perú Metazoan parasites of the minor stardrum, Stellifer minor (Tschudi (Osteichthyes, Sciaenidae, caught by artisanal fishery on Chorrillos, Lima, Peru

    Directory of Open Access Journals (Sweden)

    José Iannacone

    2004-12-01

    Full Text Available Se investigaron algunos componentes comunitarios de la parasitofauna de 105 ejemplares de Stellifer minor (Tschudi, 1844 colectados del Terminal Pesquero de Chorrillos, Lima, Perú, entre el mayo y octubre de 1998 y necropsiados para estudiar sus comunidades parasitarias. De los peces colectados, 71 fueron machos y 34 hembras. Los peces mostraron una longitud estándar entre 10,20-20,50 cm (promedio = 15,50 ± 1,65. Los parásitos metazoos fueron colectados y censados empleando las técnicas convencionales. Se colectaron un total de 3483 especimenes durante todo el muestreo, con una abundancia media total de 33,17 (3-122. El promedio de la riqueza de especies de parásitos fue 1,9 (1-4. Un hospedero no presentó ningún parásito. 20 hospederos (19,04% mostraron infección con un solo parásito, 77 (73,33% y 7 (6,66% tuvieron una infección múltiple, con 2 y 3 especies de parásitos, respectivamente. Se encontraron cinco parásitos: Rhamnocercus oliveri Luque & Iannacone, 1991 y R. stelliferi Luque & Iannacone, 1991 (Monogenea (prevalencia = 98,09%; intensidad media = 28,85; abundancia media = 28,58, Clavellotis dilatata (Kroyer, 1863 (Copepoda (prevalencia = 2,85%; intensidad media = 1; abundancia media = 0,02, Helicometra fasciata (Rudolphi, 1819 (Digenea (prevalencia = 79,04%; intensidad media = 5,77; abundancia media = 4,56 y Procamallanus (Spirocamallanus pereirai Annereaux, 1946 (Nematoda (prevalencia = 4,76%; intensidad media = 1,6; abundancia media = 0,07. Se encontró efecto del sexo en la intensidad y abundancia media de Infección de Rhamnocercus Monaco, Wood & Mizelle, 1954 y también efecto del sexo con la abundancia media de infección con H. fasciata. La diversidad media de las infracomunidades según el índice de Shannon-Weaver de S. minor fue (H' = 0,11 y el índice de Simpson (C = 0,98. Se compararon los resultados obtenidos con la estructura comunitaria parasitaria registrada en la década anterior en S. minor en la misma

  11. GPR Technologies and Methodologies in Italy: A Review

    Science.gov (United States)

    Benedetto, Andrea; Frezza, Fabrizio; Manacorda, Guido; Massa, Andrea; Pajewski, Lara

    2014-05-01

    . Frezza, P. Nocito, L. Porrini, L. Pajewski, G. Schettini e C. Twizere, 'FDTD Simulation of GPR Measurements in a Laboratory Sandbox for Landmine Detection', Proc. IWAGPR 2009, Granada, Spagna, 27-29 maggio 2009, pp. 45-49. [2] M. Balsi, S. Esposito, F. Frezza, P. Nocito, P. M. Barone, S. E. Lauro, E. Mattei, E. Pettinelli, G. Schettini e C. Twizere, 'GPR Measurements and FDTD Simulations for Landmine Detection', Proc. XIII International Conference on Ground Penetrating Radar, 21-25 giugno 2010, Lecce, pp. 865-869. [3] M. Balsi, P. M. Barone, S. Esposito, F. Frezza, S. E. Lauro, P. Nocito, E. Pettinelli, G. Schettini e C. Twizere, 'FDTD Simulations and GPR Measurements for Land Mine Detection in a Controlled Environment', Atti XVIII Riunione Nazionale di Elettromagnetismo, Benevento, 6-10 settembre 2010, pp. 59-65. [4] M. Salucci, D. Sartori, N. Anselmi, A. Randazzo, G. Oliveri, and A. Massa, 'Imaging buried objects within the second-order born approximation through a multiresolution regularized Inexact-Newton method', 2013 International Symposium on Electromagnetic Theory (EMTS), (Hiroshima, Japan), May 20-24 2013. [5] L. Lizzi, F. Viani, P. Rocca, G. Oliveri, M. Benedetti and A. Massa, 'Three-dimensional real-time localization of subsurface objects - From theory to experimental validation,' 2009 IEEE International Geoscience and Remote Sensing Symposium, vol. 2, pp. II-121-II-124, 12-17 July 2009. [6] S. Meschino, L. Pajewski, M. Pastorino, A. Randazzo, and G. Schettini, 'Detection of Subsurface Metallic Utilities by Means of a SAP Technique: Comparing MUSIC- and SVM-Based Approaches', J. Appl. Geophy., vol. 97, pp. 60-68, Oct. 2013. [7] M. Pastorino, and A. Randazzo, 'Buried object detection by an Inexact-Newton applied to nonlinear inverse scattering', Int. J. of Microwave Sci. Technol., vol. 2012, Article ID 637301, 7 pages, 2012. [8] F. Soldovieri and L. Crocco, "Electromagnetic Tomography", in Subsurface Sensing, A. S. Turk, K. A. Hocaoglu, A. A. Vertiy ed., Wiley

  12. Pms2 and uracil-DNA glycosylases act jointly in the mismatch repair pathway to generate Ig gene mutations at A-T base pairs.

    Science.gov (United States)

    Girelli Zubani, Giulia; Zivojnovic, Marija; De Smet, Annie; Albagli-Curiel, Olivier; Huetz, François; Weill, Jean-Claude; Reynaud, Claude-Agnès; Storck, Sébastien

    2017-04-03

    During somatic hypermutation (SHM) of immunoglobulin genes, uracils introduced by activation-induced cytidine deaminase are processed by uracil-DNA glycosylase (UNG) and mismatch repair (MMR) pathways to generate mutations at G-C and A-T base pairs, respectively. Paradoxically, the MMR-nicking complex Pms2/Mlh1 is apparently dispensable for A-T mutagenesis. Thus, how detection of U:G mismatches is translated into the single-strand nick required for error-prone synthesis is an open question. One model proposed that UNG could cooperate with MMR by excising a second uracil in the vicinity of the U:G mismatch, but it failed to explain the low impact of UNG inactivation on A-T mutagenesis. In this study, we show that uracils generated in the G1 phase in B cells can generate equal proportions of A-T and G-C mutations, which suggests that UNG and MMR can operate within the same time frame during SHM. Furthermore, we show that Ung -/- Pms2 -/- mice display a 50% reduction in mutations at A-T base pairs and that most remaining mutations at A-T bases depend on two additional uracil glycosylases, thymine-DNA glycosylase and SMUG1. These results demonstrate that Pms2/Mlh1 and multiple uracil glycosylases act jointly, each one with a distinct strand bias, to enlarge the immunoglobulin gene mutation spectrum from G-C to A-T bases. © 2017 Girelli Zubani et al.

  13. Tiger Rag na interpretação do Le Quintette du Hot Club de France: história, análise e práticas de performance Tiger Rag as performed by the Quintet of the Hot Club of France: history, analysis and performance practices

    Directory of Open Access Journals (Sweden)

    Adriana Costa

    2011-06-01

    Full Text Available Estudo sobre a assimilação do jazz na cultura popular francesa dos anos de 1930 e criação de uma sociedade de entusiastas desse estilo, especialmente em torno do Le Quintette du Hot Club de France. A transcrição e análise de Tiger Rag, música dos membros da Original Dixieland Jazz Band (Nick La Rocca, Eddie Edwards, Tony Sbarbaro, Larry Shields e Harry da Costa na interpretação do Le Quintette (LE QUINTETTE DU HOT CLUB DE FRANCE, 1934, remasterizado em 1993 revela suas práticas de performance, especialmente de seus solistas: Django Reinhardt, no violão, e Stephane Grappelli, no violino.Study about the assimilation of jazz into French popular culture of the 1930s and the emergence of a society of jazz enthusiasts, especially around Le Quintette du Hot Club de France. The transcription and analysis of Tiger Rag, composed by the members of the Original Dixieland Jazz Band (Nick La Rocca, Eddie Edwards, Tony Sbarbaro, Larry Shields and Harry da Costa and performed by Le Quintette reveal performance practices of its main soloists: Django Reinhardt on the guitar and Stephane Grappelli on the violin.

  14. Palindromic Molecule Beacon-Based Cascade Amplification for Colorimetric Detection of Cancer Genes.

    Science.gov (United States)

    Shen, Zhi-Fa; Li, Feng; Jiang, Yi-Fan; Chen, Chang; Xu, Huo; Li, Cong-Cong; Yang, Zhe; Wu, Zai-Sheng

    2018-03-06

    A highly sensitive and selective colorimetric assay based on a multifunctional molecular beacon with palindromic tail (PMB) was proposed for the detection of target p53 gene. The PMB probe can serve as recognition element, primer, and polymerization template and contains a nicking site and a C-rich region complementary to a DNAzyme. In the presence of target DNA, the hairpin of PMB is opened, and the released palindromic tails intermolecularly hybridize with each other, triggering the autonomous polymerization/nicking/displacement cycles. Although only one type of probe is involved, the system can execute triple and continuous polymerization strand displacement amplifications, generating large amounts of G-quadruplex fragments. These G-rich fragments can bind to hemin and form the DNAzymes that possess the catalytic activity similar to horseradish peroxidase, catalyzing the oxidation of ABTS by H 2 O 2 and producing the colorimetric signal. Utilizing the newly proposed sensing system, target DNA can be detected down to 10 pM with a linear response range from 10 pM to 200 nM, and mutant target DNAs are able to be distinguished even by the naked eye. The desirable detection sensitivity, high specificity, and operation convenience without any separation step and chemical modification demonstrate that the palindromic molecular beacon holds the potential for detecting and monitoring a variety of nucleic acid-related biomarkers.

  15. Physical association of pyrimidine dimer DNA glycosylase and apurinic/apyrimidinic DNA endonuclease essential for repair of ultraviolet-damaged DNA

    International Nuclear Information System (INIS)

    Nakabeppu, Y.; Sekiguchi, M.

    1981-01-01

    T4 endonuclease, which is involved in repair of uv-damaged DNA, has been purified to apparent physical homogeneity. Incubation of uv-irradiated poly(dA).poly(dT) with the purified enzyme preparations resulted in production of alkali-labile apyrimidinic sites, followed by formation of nicks in the polymer. By performing a limited reaction with T4 endonuclease V at pH 8.5, irradiated polymer was converted to an intermediate form that carried a large number of alkali-labile sites but only a few nicks. The intermediate was used as substrate for the assay of apurinic/apyrimidinic DNA endonuclease activity. The two activities, a pyrimidine dimer DNA glycosylase and an apurinic/apyrimidinic DNA endonuclease, were copurified and found in enzyme preparations that contained only a 16,000-dalton polypeptide. These results strongly suggested that a DNA glycosylase specific for pyrimidine dimers and an apurinic/apyrimidinic DNA endonuclease reside in a single polypeptide chain coded by the denV gene of bacteriophage T4

  16. Functional recovery in rat spinal cord injury induced by hyperbaric oxygen preconditioning.

    Science.gov (United States)

    Lu, Pei-Gang; Hu, Sheng-Li; Hu, Rong; Wu, Nan; Chen, Zhi; Meng, Hui; Lin, Jiang-Kai; Feng, Hua

    2012-12-01

    It is a common belief that neurosurgical interventions can cause inevitable damage resulting from the procedure itself in surgery especially for intramedullary spinal cord tumors. The present study was designed to examine if hyperbaric oxygen preconditioning (HBO-PC) was neuroprotective against surgical injuries using a rat model of spinal cord injury (SCI). Sprague-Dawley rats were randomly divided into three groups: HBO-PC group, hypobaric hypoxic preconditioning (HH-PC) control group, and normobaric control group. All groups were subjected to SCI by weight drop device. Rats from each group were examined for neurological behavior and electrophysiological function. Tissue sections were analyzed by using immunohistochemistry, TdT-mediated dUTP-biotin nick end labeling, and axonal tract tracing. Significant neurological deficits were observed after SCI and HBO-PC and HH-PC improved neurological deficits 1 week post-injury. The latencies of motor-evoked potential and somatosensory-evoked potential were significantly delayed after SCI, which was attenuated by HBO-PC and HH-PC. Compared with normobaric control group, pretreatment with HBO and hypobaric hypoxia significantly reduced the number of TdT-mediated dUTP-biotin nick end labeling-positive cells, and increased nestin-positive cells. HBO-PC and HH-PC enhanced axonal growth after SCI. In conclusion, preconditioning with HBO and hypobaric hypoxia can facilitate functional recovery and suppress cell apoptosis after SCI and may prove to be a useful preventive strategy to neurosurgical SCI.

  17. DNA Packaging Specificity of Bacteriophage N15 with an Excursion into the Genetics of a Cohesive End Mismatch.

    Directory of Open Access Journals (Sweden)

    Michael Feiss

    Full Text Available During DNA replication by the λ-like bacteriophages, immature concatemeric DNA is produced by rolling circle replication. The concatemers are processed into mature chromosomes with cohesive ends, and packaged into prohead shells, during virion assembly. Cohesive ends are generated by the viral enzyme terminase, which introduces staggered nicks at cos, an approx. 200 bp-long sequence containing subsites cosQ, cosN and cosB. Interactions of cos subsites of immature concatemeric DNA with terminase orchestrate DNA processing and packaging. To initiate DNA packaging, terminase interacts with cosB and nicks cosN. The cohesive ends of N15 DNA differ from those of λ at 2/12 positions. Genetic experiments show that phages with chromosomes containing mismatched cohesive ends are functional. In at least some infections, the cohesive end mismatch persists through cyclization and replication, so that progeny phages of both allelic types are produced in the infected cell. N15 possesses an asymmetric packaging specificity: N15 DNA is not packaged by phages λ or 21, but surprisingly, N15-specific terminase packages λ DNA. Implications for genetic interactions among λ-like bacteriophages are discussed.

  18. Change at the top for the CERN physics schools

    CERN Multimedia

    2009-01-01

    After directing the CERN physics schools since 1993, Egil Lillestøl has handed over to Nick Ellis. At the same time, Hélène Haller has taken over from Danielle Métral as the schools’ administrator. From left to right: Nick Ellis, Hélène Haller, Danielle Métral and Egil Lillestøl.The CERN physics schools for young experimentalists date back to the 1960s and as early as 1971 collaboration with the Joint Institute for Nuclear Research (JINR) in Dubna led to the Joint CERN-JINR schools, which reached beyond CERN’s Member States every two years. Then, in 1993, CERN and JINR agreed to organize the schools jointly every year, as the European School for High-Energy Physics. Egil Lillestøl, has not only run this school very successfully since then, but also created the CERN Latin American School of High-Energy Physics, beginning in 2001. Danielle Métral has been responsible for the schools’ administration since 2001, both for t...

  19. Radioactive probes for human gene localisation by in situ hybridisation

    International Nuclear Information System (INIS)

    Fennell, S.J.

    1980-07-01

    Radioactive probes of high specific activity have been used for human gene localisation on metaphase chromosome preparations. Human 5S ribosomal RNA was used as a model system, as a probe for the localisation of human 5S ribosomal genes. 125 I-labelled mouse 5S ribosomal RNA was used to study the 5S ribosomal gene content and arrangement in families with translocations on the long arm of chromosome 1 close to or containing the 5S ribosomal RNA locus, by in situ hybridisation to human metaphase chromosomes from peripheral blood cultures. This confirmed the chromosomal assignment of 5S ribosomal genes to 1q 42-43. In situ hybridisation probes were also prepared from recombinant plasmids containing Xenopus laevis oocyte 5S or 28S/18S gene sequences to give [ 3 H]-labelled cRNA and [ 3 H]-labelled nick-translated plasmid DNA. Studies on the kinetics of hybridisation of plasmid probes with and without ribosomal gene sequences questioned the role of plasmid DNA for amplification of signal during gene localisation. Gene localisation was obtained with nick-translated plasmid DNA containing the 28S/18S ribosomal DNA insert after short exposure times, but poor results were obtained using a [ 3 H]-labelled cRNA probe transcribed from the plasmid with the 5S gene insert. (author)

  20. Dual-cyclical nucleic acid strand-displacement polymerization based signal amplification system for highly sensitive determination of p53 gene.

    Science.gov (United States)

    Xu, Jianguo; Wu, Zai-Sheng; Li, Hongling; Wang, Zhenmeng; Le, Jingqing; Zheng, Tingting; Jia, Lee

    2016-12-15

    In the present study, we proposed a novel dual-cyclical nucleic acid strand-displacement polymerization (dual-CNDP) based signal amplification system for highly sensitive determination of tumor suppressor genes. The system primarily consisted of a signaling hairpin probe (SHP), a label-free hairpin probe (LHP) and an initiating primer (IP). The presence of target DNA was able to induce one CNDP through continuous process of ligation, polymerization and nicking, leading to extensively accumulation of two nicked triggers (NT1 and NT2). Intriguingly, the NT1 could directly hybridize SHP, while the NT2 could act as the target analog to induce another CNDP. The resulting dual-CNDP contributed the striking signal amplification, and only a very weak blank noise existed since the ligation template of target was not involved. In this case, the target could be detected in a wide linear range (5 orders of magnitude), and a low detection limit (78 fM) was obtained, which is superior to most of the existing fluorescent methods. Moreover, the dual-CNDP sensing system provided a high selectivity towards target DNA against mismatched target and was successfully applied to analysis of target gene extracted from cancer cells or in human serum-contained samples, indicating its great potential for practical applications. Copyright © 2016 Elsevier B.V. All rights reserved.

  1. The Study of Phenological and Morphophysiological Traits of New and Old Barely Shoot Cultivars

    Directory of Open Access Journals (Sweden)

    N Moshfeghi

    2015-07-01

    Full Text Available In order to compare morphophysiological traits between new and old cultivars of barley (Hordeum vulgare L. a greenhouse experiment was conducted in greenhouse condition in Faculty of Agriculture, Ferdowsi University of Mashhad in spring and summer 2011.The type of experiment was a completely randomized design with eight treatments (Nosrat , Youssof , Fagr30 , Nick and four old cultivars include (Reihan, Kavir, Zarjo , Valfajr and three replications. All seeds were grown in PVC tubes contained washed sands and disruptive sampling was occurred with destroying plastic bags in the end of the four stages uncluding 6th leaf stage, jointing, heading and ripening. Results indicated that new cultivars showed better performances in all shoot growth parameters than the old cultivars. Modern cultivars for most traits, particularly traits that were positively correlated with yield and its components proved more successful than the old cultivars. They had longer reproductive periods and grain filling duration but their vegetative period were shorter than old cultivars. Among shoots characteristics, new cultivars had higher leaf area, stem dry weight and stem length than old cultivars. Result of this study showed that, among all the characteristics of grain yield and yield components, such as grain weight ,spike length, spike per plant, grain per spike and number tiller , new figures were better than old figures. In general, new cultivar Nick was introduced as the best cultivar and Zarjo was introduced as the weakest cultivar.

  2. Quantification and genome-wide mapping of DNA double-strand breaks.

    Science.gov (United States)

    Grégoire, Marie-Chantal; Massonneau, Julien; Leduc, Frédéric; Arguin, Mélina; Brazeau, Marc-André; Boissonneault, Guylain

    2016-12-01

    DNA double-strand breaks (DSBs) represent a major threat to the genetic integrity of the cell. Knowing both their genome-wide distribution and number is important for a better assessment of genotoxicity at a molecular level. Available methods may have underestimated the extent of DSBs as they are based on markers specific to those undergoing active repair or may not be adapted for the large diversity of naturally occurring DNA ends. We have established conditions for an efficient first step of DNA nick and gap repair (NGR) allowing specific determination of DSBs by end labeling with terminal transferase. We used DNA extracted from HeLa cells harboring an I-SceI cassette to induce a targeted nick or DSB and demonstrated by immunocapture of 3'-OH that a prior step of NGR allows specific determination of loci-specific or genome wide DSBs. This method can be applied to the global determination of DSBs using radioactive end labeling and can find several applications aimed at understanding the distribution and kinetics of DSBs formation and repair. Copyright © 2016 Elsevier B.V. All rights reserved.

  3. Factors affecting mutational specificity in mammalian cells: [Informal technical progress report], February 1, 1986-January 31, 1987

    International Nuclear Information System (INIS)

    Strauss, B.S.

    1987-01-01

    We analyzed the different c-H-ras mutations produced in cells after treatment with chemical carcinogens. The overall goal of this work is an understanding of the changes produced by environmental mutagens and carcinogens to learn how closely the results obtained with bacterial and cellular assay systems apply to the in vivo situation. Our work continues to utilize the technique for the study of DNA synthesis termination in vitro developed in this laboraory and utilized for the past several years. This technology can be characterized as a Sanger dideoxy sequencing technique with mutation induced lesions in the template strand serving as chain terminators instead of the dideoxynucleotides used in the Sanger technique to terminate the growing strand. We have added to this technique a modification for the study of termination on double stranded templates as possibly modeling more closely the natural situation. For this modification, double stranded M13 DNA is split once with a restriction enzyme which makes a unique cut and the linearized DNA is hybridized with single stranded circular DNA to make a uniquely nicked double stranded circle with a single 3'OH to serve as a primer for DNA synthesis which can occur by either strand displacement or nick translation

  4. Distinct kinetics of human DNA ligases I, IIIalpha, IIIbeta, and IV reveal direct DNA sensing ability and differential physiological functions in DNA repair

    Energy Technology Data Exchange (ETDEWEB)

    Chen, Xi; Ballin, Jeff D.; Della-Maria, Julie; Tsai, Miaw-Sheue; White, Elizabeth J.; Tomkinson, Alan E.; Wilson, Gerald M.

    2009-05-11

    The three human LIG genes encode polypeptides that catalyze phosphodiester bond formation during DNA replication, recombination and repair. While numerous studies have identified protein partners of the human DNA ligases (hLigs), there has been little characterization of the catalytic properties of these enzymes. In this study, we developed and optimized a fluorescence-based DNA ligation assay to characterize the activities of purified hLigs. Although hLigI joins DNA nicks, it has no detectable activity on linear duplex DNA substrates with short, cohesive single-strand ends. By contrast, hLigIII{beta} and the hLigIII{alpha}/XRCC1 and hLigIV/XRCC4 complexes are active on both nicked and linear duplex DNA substrates. Surprisingly, hLigIV/XRCC4, which is a key component of the major non-homologous end joining (NHEJ) pathway, is significantly less active than hLigIII on a linear duplex DNA substrate. Notably, hLigIV/XRCC4 molecules only catalyze a single ligation event in the absence or presence of ATP. The failure to catalyze subsequent ligation events reflects a defect in the enzyme-adenylation step of the next ligation reaction and suggests that, unless there is an in vivo mechanism to reactivate DNA ligase IV/XRCC4 following phosphodiester bond formation, the cellular NHEJ capacity will be determined by the number of adenylated DNA ligaseIV/XRCC4 molecules.

  5. Human DNA ligase III bridges two DNA ends to promote specific intermolecular DNA end joining

    Science.gov (United States)

    Kukshal, Vandna; Kim, In-Kwon; Hura, Gregory L.; Tomkinson, Alan E.; Tainer, John A.; Ellenberger, Tom

    2015-01-01

    Mammalian DNA ligase III (LigIII) functions in both nuclear and mitochondrial DNA metabolism. In the nucleus, LigIII has functional redundancy with DNA ligase I whereas LigIII is the only mitochondrial DNA ligase and is essential for the survival of cells dependent upon oxidative respiration. The unique LigIII zinc finger (ZnF) domain is not required for catalytic activity but senses DNA strand breaks and stimulates intermolecular ligation of two DNAs by an unknown mechanism. Consistent with this activity, LigIII acts in an alternative pathway of DNA double strand break repair that buttresses canonical non-homologous end joining (NHEJ) and is manifest in NHEJ-defective cancer cells, but how LigIII acts in joining intermolecular DNA ends versus nick ligation is unclear. To investigate how LigIII efficiently joins two DNAs, we developed a real-time, fluorescence-based assay of DNA bridging suitable for high-throughput screening. On a nicked duplex DNA substrate, the results reveal binding competition between the ZnF and the oligonucleotide/oligosaccharide-binding domain, one of three domains constituting the LigIII catalytic core. In contrast, these domains collaborate and are essential for formation of a DNA-bridging intermediate by adenylated LigIII that positions a pair of blunt-ended duplex DNAs for efficient and specific intermolecular ligation. PMID:26130724

  6. A Graphene-Based Biosensing Platform Based on Regulated Release of an Aptameric DNA Biosensor.

    Science.gov (United States)

    Mao, Yu; Chen, Yongli; Li, Song; Lin, Shuo; Jiang, Yuyang

    2015-11-09

    A novel biosensing platform was developed by integrating an aptamer-based DNA biosensor with graphene oxide (GO) for rapid and facile detection of adenosine triphosphate (ATP, as a model target). The DNA biosensor, which is locked by GO, is designed to contain two sensing modules that include recognition site for ATP and self-replication track that yields the nicking domain for Nt.BbvCI. By taking advantage of the different binding affinity of single-stranded DNA, double-stranded DNA and aptamer-target complex toward GO, the DNA biosensor could be efficiently released from GO in the presence of target with the help of a complementary DNA strand (CPDNA) that partially hybridizes to the DNA biosensor. Then, the polymerization/nicking enzyme synergetic isothermal amplification could be triggered, leading to the synthesis of massive DNA amplicons, thus achieving an enhanced sensitivity with a wide linear dynamic response range of four orders of magnitude and good selectivity. This biosensing strategy expands the applications of GO-DNA nanobiointerfaces in biological sensing, showing great potential in fundamental research and biomedical diagnosis.

  7. Genomic Mapping of Human DNA provides Evidence of Difference in Stretch between AT and GC rich regions

    Science.gov (United States)

    Reifenberger, Jeffrey; Dorfman, Kevin; Cao, Han

    Human DNA is a not a polymer consisting of a uniform distribution of all 4 nucleic acids, but rather contains regions of high AT and high GC content. When confined, these regions could have different stretch due to the extra hydrogen bond present in the GC basepair. To measure this potential difference, human genomic DNA was nicked with NtBspQI, labeled with a cy3 like fluorophore at the nick site, stained with YOYO, loaded into a device containing an array of nanochannels, and imaged. Over 473,000 individual molecules of DNA, corresponding to roughly 30x coverage of a human genome, were collected and aligned to the human reference. Based on the known AT/GC content between aligned pairs of labels, the stretch was measured for regions of similar size but different AT/GC content. We found that regions of high GC content were consistently more stretched than regions of high AT content between pairs of labels varying in size between 2.5 kbp and 500 kbp. We measured that for every 1% increase in GC content there was roughly a 0.06% increase in stretch. While this effect is small, it is important to take into account differences in stretch between AT and GC rich regions to improve the sensitivity of detection of structural variations from genomic variations. NIH Grant: R01-HG006851.

  8. A cascade autocatalytic strand displacement amplification and hybridization chain reaction event for label-free and ultrasensitive electrochemical nucleic acid biosensing.

    Science.gov (United States)

    Chen, Zhiqiang; Liu, Ying; Xin, Chen; Zhao, Jikuan; Liu, Shufeng

    2018-04-23

    Herein, an autocatalytic strand displacement amplification (ASDA) strategy was proposed for the first time, which was further ingeniously coupled with hybridization chain reaction (HCR) event for the isothermal, label-free and multiple amplification toward nucleic acid detection. During the ASDA module, the target recognition opens the immobilized hairpin probe (IP) and initiates the annealing of the auxiliary DNA strand (AS) with the opened IP for the successive polymerization and nicking reaction in the presence of DNA polymerase and nicking endonuclease. This induces the target recycling and generation of a large amount of intermediate DNA sequences, which can be used as target analogy to execute the autocatalytic strand displacement amplification. Simultaneously, the introduced AS strand can propagate the HCR between two hairpins (H1 and H2) to form a linear DNA concatamer with cytosine (C)-rich loop region, which can facilitate the in-situ synthesis of silver nanoclusters (AgNCs) as electrochemical tags for further amplification toward target responses. With current cascade ASDA and HCR strategy, the detection of target DNA could be achieved with a low detection limit of about 0.16 fM and a good selectivity. The developed biosensor also exhibits the distinct advantages of flexibility and simplicity in probe design and biosensor fabrication, and label-free electrochemical detection, thus opens a promising avenue for the detection of nucleic acid with low abundance in bioanalysis and clinical biomedicine. Copyright © 2018 Elsevier B.V. All rights reserved.

  9. Evaluación de métodos de marcación-radioactivos y no radioactivos empleados en la elaboración de sondas diagnósticas de malaria

    Directory of Open Access Journals (Sweden)

    Gladys Pinilla

    1993-09-01

    Full Text Available Se probaron métodos de marcación radioactivos dentro de los cuales el sistema Random Primers resultó mejor que el de Nick Translation. Se obtuvieron sondas con mayor actividad especifica (1-2X1O9dpmIug y una sensibilidad 10 veces mayor en la detección de ADN de Plasmodium falciparum. La sensibilidad alcanzada (12pg obtenida con la sonda PFCOL692 desarrollada y caracterizada en nuestro laboratorio, fue igual a la obtenida con la sonda pRepHind, una de las más utilizadas en los laboratorios de otros países. El método de marcación no radioactivoconfotobiotina mostró ventajas sobre la marcación por Nick Translation con nucleótidos biotinilados y se alcanzó una sensibilidad de 135 pg de ADN de P. falciparum. También se probó un sistema degeneración de señales fluorescentes mediante el empleo de proteasas, y los resultados preliminares muestran sus posibilidades como método de detección.

  10. Localization Strategies in WSNs as applied to Landslide Monitoring (Invited)

    Science.gov (United States)

    Massa, A.; Robol, F.; Polo, A.; Giarola, E.; Viani, F.

    2013-12-01

    represent a fine tuning of large scale satellite acquisitions, usually adopted for remote sensing of landslides. The integration of dense and frequent WSN data within satellite image analysis will enhance the sensing capabilities leading to a multi-resolution and an highly space-time calibrated system. The WSN-based system has been preliminary tested in controlled environments in the ELEDIA laboratories and is now installed in a real test site where an active landslide is evolving. Preliminary data are here presented to assess the feasibility of the investigated solution in landslide monitoring and event forecasting. REFERENCES [1] M. Benedetti, L. Ioriatti, M. Martinelli, and F. Viani, 'Wireless sensor network: a pervasive technology for earth observation,' in IEEE Journal of Selected Topics in App. Earth Obs. And Remote Sens., vol. 3, no. 4, pp. 488-497, 2010. [2] F. Viani, M. Donelli, P. Rocca, G. Oliveri, D. Trinchero, and A. Massa, 'Localization, tracking and imaging of targets in wireless sensor networks,' Radio Science, vol. 46, no. 5, 2011. [3] F. Viani, F. Robol, M. Salucci, E. Giarola, S. De Vigili, M. Rocca, F. Boldrini, G. Benedetti, and A. Massa, 'WSN-based early alert system for preventing wildlife-vehicle collisions in Alps regions - From the laboratory test to the real-world implementation,' 7th European Conference on Antennas and Propagation 2013 (EUCAP2013), Gothenburg, Sweden, April 8-12, 2013.

  11. In vitro analysis of radioprotective effect of monoterpenes

    International Nuclear Information System (INIS)

    Ken-ichi Kudo; Tadashi Hanafusa; Toshiro Ono

    2017-01-01

    Monoterpenes are naturally occurring hydrocarbons composed of two units of isoprenes. They exhibit antioxidant activity to scavenge reactive oxygen species, such as hydroxyl radicals. We investigated the potential of monoterpenes such as thymol, linalool, and menthol to act as radioprotectants. The proliferation of EL4 cells, a mouse lymphoma cell line, treated with linalool at a concentration of 500 μM or more was not affected by X-ray irradiation. Plasmid-nicking assay performed using formamidopyrimidine-DNA glycosylase showed that linalool prevented single strand breaks and oxidized purines on pUC19 plasmid DNA. These findings indicate that linalool has the ability to scavenge reactive oxygen species and is a potential radioprotector. (author)

  12. Separation of DNA-dependent polymerate activities in Micrococcus radiodurans

    International Nuclear Information System (INIS)

    Kitayama, S.; Matsuyama, A.

    1977-01-01

    DNA polymerase activities in Micrococcus radiodurans were separated into two fractions after purification more than 2000 fold. They differ in pH optimum and residual activities in the absence of a full deoxyribonucleoside triphosphates complement. NAD partly inhibited one of the activities. Both activities were eluted as a single peak on gel filtration and sedimented at the same rate on glycerol gradient centrifugation. Molecular weight 140000 was calculated from Stokes radius and sedimentation constant. Deoxyribonuclease activity was detected on one of the polymerase activities which preferentially degraded double-stranded DNA. Priming activity of nicked DNA was reduced by γ-radiation. These results have been related to the possible roles in repair synthesis in vivo or DNA synthesis in permeable cells of M. radiodurans

  13. Three additional genes involved in pyrimidine dimer removal in Saccharomyces cerevisiae: RAD7, RAD14, and MMS19

    Energy Technology Data Exchange (ETDEWEB)

    Prakash, L; Prakash, S

    1979-01-01

    The ability to remove ultraviolet (uv)-induced pyrimidine dimers from the nuclear DNA of yeast was examined in two radiation-sensitive (rad) mutants and one methyl methanesulfonate-sensitive (mms) mutant of the yeast Saccharomyces cerevisiae. The susceptibility of DNA from irradiated cells to nicking by an endonuclease activity prepared from crude extracts of Micrococcus luteus was used to measure the presence of dimers in DNA. The rad7, rad14, and mms19 mutants were found to be defective in their ability to remove uv-induced dimers from nuclear DNA. All three mutants belong to the same episatic group as the other mutants involved in excision-repair. All three mutants show enhanced uv-induced mutations. The rad 14 mutant also shows epistatic interactions with genes in the other two uv repair pathways.

  14. Separation of DNA-dependent polymerase activities in Micrococcus radiodurans

    Energy Technology Data Exchange (ETDEWEB)

    Kitayama, S; Matsuyama, A [Institute of Physical and Chemical Research, Wako, Saitama (Japan)

    1977-03-02

    DNA polymerase activities in Micrococcus radiodurans were separated into two fractions after purification more than 2000 fold. They differ in pH optimum and residual activities in the absence of a full deoxyribonucleoside triphosphates complement. NAD partly inhibited one of the activities. Both activities were eluted as a single peak on gel filtration and sedimented at the same rate on glycerol gradient centrifugation. Molecular weight 140000 was calculated from Stokes radius and sedimentation constant. Deoxyribonuclease activity was detected on one of the polymerase activities which preferentially degraded double-stranded DNA. Priming activity of nicked DNA was reduced by ..gamma.. radiation. These results have been related to the possible roles in repair synthesis in vivo or DNA synthesis in permeable cells of M. radiodurans.

  15. Expression analysis of a ''Cucurbita'' cDNA encoding endonuclease

    International Nuclear Information System (INIS)

    Szopa, J.

    1995-01-01

    The nuclear matrices of plant cell nuclei display intrinsic nuclease activity which consists in nicking supercoiled DNA. A cDNA encoding a 32 kDa endonuclease has been cloned and sequenced. The nucleotide and deduced amino-acid sequences show high homology to known 14-3-3-protein sequences from other sources. The amino-acid sequence shows agreement with consensus sequences for potential phosphorylation by protein kinase A and C and for calcium, lipid and membrane-binding sites. The nucleotide-binding site is also present within the conserved part of the sequence. By Northern blot analysis, the differential expression of the corresponding mRNA was detected; it was the strongest in sink tissues. The endonuclease activity found on DNA-polyacrylamide gel electrophoresis coincided with mRNA content and was the highest in tuber. (author). 22 refs, 6 figs

  16. Programmed cell death of tobacco BY-2 cells induced by still culture conditions is affected by the age of the culture under agitation.

    Science.gov (United States)

    Hiraga, Asahi; Kaneta, Tsuyoshi; Sato, Yasushi; Sato, Seiichi

    2010-01-25

    Evans Blue staining indicated that actively growing tobacco BY-2 cells in the exponential phase died more rapidly than quiescent cells in the stationary phase when the cells cultured under agitation were placed under still conditions. Fifty percent cell death was induced at about 18, 26, 80 and 140 h for early, mid, late exponential- and stationary-phase cells, respectively. Actively growing cells became TUNEL (transferase-mediated dUTP nick end labelling)-positive more rapidly than quiescent cells, suggesting that the cell death evaluated by Evans Blue is accompanied by DNA cleavages. Electrophoresis of genomic DNA showed a typical 'DNA laddering' pattern formed by multiples of about 200 bp internucleosomal units. Chromatin condensation was first detected at least within 24 h by light microscopy, and then cell shrinkage followed. These findings suggest that the death of BY-2 cells induced by still conditions is PCD (programmed cell death).

  17. Characteristics of palindromic sequences in DNA of the sea urchin Stronglyocentrotus intermedius

    International Nuclear Information System (INIS)

    Brykov, V.A.; Kukhlevskii, A.D.

    1986-01-01

    The fraction of palindromic sequences in the nuclear DNA of the sea urchin S. intermedius was characterized. Using chromatography on hydroxyapatite and treatment with S1 nuclease, it was shown that the fraction of palindromic sequences more than doubles when the sodium concentration in solution is increased or the temperature of reassociation is lowered. The increase is due to the involvement of inverted repeats in reassociation, which are characterized by a substantial nonhomologous character and/or the presence of an extended intervening DNA sequence. It was found by the method of reassociation of a nicked palindrome fraction with an excess of total homologous DNA that most of the inverted repeats in the sea urchin genome are unique sequences. The complexity of the palindrome fraction was estimated at 8.2 x 10 7 nucleotide pairs, and the number of palindromes per haploid genome ∼ 500,000

  18. Generation of Humanized Mouse Models with Focus on Antithrombin Deficiency

    DEFF Research Database (Denmark)

    Jensen, Astrid Bøgh

    2015-01-01

    transgene. The CRISPR/Cas9 system is a relatively new and innovative method for targeted mutagenesis. The Cas9 nuclease introduces a double stranded break in the DNA, which can be repaired through homologous recombination of a targeting vector. A mutated Cas9n (Cas9 nickase) has been designed, which only...... gene by the murine antithrombin regulatory sequences, I designed a targeted mutagenesis using the CRISPR/Cas9 system which conserves the 5’UTR of the murine antithrombin gene. With the CRISPR/Cas9 I achieved targeting efficiency for heterozygous integrations of about 80%, which correlated well with our...... preliminary CRISPR/Cas9 experiments targeting the Rosa26 locus. However, when targeting the Rosa26 locus, using the CRISPR/Cas9n system I only observed 65% targeting efficiency for heterozygous integration which correlates well with the requirement for two nicks created by the mutated Cas9n. Others have shown...

  19. Accelerator design and construction in the 1950s

    International Nuclear Information System (INIS)

    Blewett, J.P.

    1989-01-01

    This article looks into the history of the design, construction and operation of four of the large particle accelerators of the 1950s, the Cosmotron and more powerful alternating-gradient synchrotron (AGS) at Brookhaven, the Bevatron at Berkeley and the CERN proton synchrotron in Geneva with which the author was involved. The author's own contribution was in magnet design for the Cosmotron and the radiofrequency accelerating system. He later worked on linear accelerators and strong focusing later used in the AGS with Nick Christofilos from Athens. Collaboration between CERN and Brookhaven continued following a British study of alternating-gradient focusing which showed up possible resonance problems. In 1953, the ''phase transition'' problem was overcome. The author's personal contribution to the AGS project completes the article. (UK)

  20. From welcome to well . come: the mobilities, temporalities and geopolitics of contemporary hospitality – commentary to Gill

    Directory of Open Access Journals (Sweden)

    Roger Norum

    2018-05-01

    Full Text Available In this reflection, I take Nick Gill’s contribution to this issue on the politics of hospitality and welcome as a point of departure to address three specific concerns, framed by the context of contemporary global mobility. First, I argue that various aspects of the recent so-called European migration crisis enable us to further question some of the long-fixed categories through which mobile actors are often classified. Second, I speak to the critical nature of temporality as a governing factor not just in how states manage mobile peoples, but in how people imagine, understand and experience individual and group processes of welcome. Finally, I suggest that just as geopolitical discourse plays a clear role in how migration is understood and experienced, so too are geopolitics deeply embedded in the encounters, practices and regimes of hospitality across tourism.

  1. DNA fragmentation in spermatozoa

    DEFF Research Database (Denmark)

    Rex, A S; Aagaard, J.; Fedder, J

    2017-01-01

    Sperm DNA Fragmentation has been extensively studied for more than a decade. In the 1940s the uniqueness of the spermatozoa protein complex which stabilizes the DNA was discovered. In the fifties and sixties, the association between unstable chromatin structure and subfertility was investigated....... In the seventies, the impact of induced DNA damage was investigated. In the 1980s the concept of sperm DNA fragmentation as related to infertility was introduced as well as the first DNA fragmentation test: the Sperm Chromatin Structure Assay (SCSA). The terminal deoxynucleotidyl transferase nick end labelling...... (TUNEL) test followed by others was introduced in the nineties. The association between DNA fragmentation in spermatozoa and pregnancy loss has been extensively investigated spurring the need for a therapeutic tool for these patients. This gave rise to an increased interest in the aetiology of DNA damage...

  2. Scientific Information Policy Board 90th Meeting

    CERN Multimedia

    AUTHOR|(CDS)2051371

    2017-01-01

    Gigi Rolandi, sitting in the middle, chairing his last meeting of the Scientific Information Policy Board before his retirement. Rolandi was the chairman of the board from March 2006 until April 2017, a period that has seen lots of dynamics within the field of authoring, publishing and librarianship - both at CERN and beyond. In the first row from left to right: Eckhard Elsen (Director of Research), Jens Vigen (Head Librarian) Gigi Rolandi (SIPB Chair), Anita Hollier (Archivist) and Urs Wiedemann (Theory). In the second row from left to right: Nikos Kasioumis (Invited speaker), Alexander Kohls (Invited speaker), Stella Christodoulaki (Invited speaker), Nick Ellis (Chair CREB), Clara Troncon (ACCU), Annette Holtkamp (Scientific Information Service), Tim Smith (IT) Brigitte Bloch-Devaux (non-LHC experiments), Constantinos Loizides (ALICE), Thierry Stora (Engineering), John Jowett (Beams), Arjan Verweij (Technology), Anne Gentil-Beccot (Scientific Information Service), Fergus Wilson (LHCb), Ludmila Marian (Invit...

  3. An episomal vector-based CRISPR/Cas9 system for highly efficient gene knockout in human pluripotent stem cells.

    Science.gov (United States)

    Xie, Yifang; Wang, Daqi; Lan, Feng; Wei, Gang; Ni, Ting; Chai, Renjie; Liu, Dong; Hu, Shijun; Li, Mingqing; Li, Dajin; Wang, Hongyan; Wang, Yongming

    2017-05-24

    Human pluripotent stem cells (hPSCs) represent a unique opportunity for understanding the molecular mechanisms underlying complex traits and diseases. CRISPR/Cas9 is a powerful tool to introduce genetic mutations into the hPSCs for loss-of-function studies. Here, we developed an episomal vector-based CRISPR/Cas9 system, which we called epiCRISPR, for highly efficient gene knockout in hPSCs. The epiCRISPR system enables generation of up to 100% Insertion/Deletion (indel) rates. In addition, the epiCRISPR system enables efficient double-gene knockout and genomic deletion. To minimize off-target cleavage, we combined the episomal vector technology with double-nicking strategy and recent developed high fidelity Cas9. Thus the epiCRISPR system offers a highly efficient platform for genetic analysis in hPSCs.

  4. Observing comets

    CERN Document Server

    James, Nick

    2003-01-01

    Since comet Shoemaker-Levy collided with the planet Jupiter with stupendous force in 1994 there has been an upsurge of amateur interest in comets Most comets are first discovered by amateur astronomers because there are so many amateurs looking for them, and techniques and instruments have improved dramatically in the past few years After a short but detailed introduction to the comets themselves Nick James and Gerald North describe comet hunting, photographing and imaging comets, and digital image processing The use of computers for orbital calculations and even helping to discover new comets is given a full chapter, as are advanced techniques including comet photometry and spectroscopy This comprehensive book has an accompanying CD-ROM and is at once a "primer" for comet hunters and a reference text for more advanced amateur astronomers

  5. Population control of resident and immigrant microglia by mitosis and apoptosis

    DEFF Research Database (Denmark)

    Wirenfeldt, Martin; Dissing-Olesen, Lasse; Babcock, Alicia

    2007-01-01

    microglia often occurred in clusters, some having recently incorporated bromodeoxyuridine, showing that proliferation had occurred. Annexin V labeling and staining for activated caspase-3 and terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling showed that apoptotic mechanisms participate...... in dissolution of the microglial response. Using bone marrow chimeric mice, we found that the lesion-induced proliferative capacity of resident microglia superseded that of immigrant microglia, whereas lesion-induced kinetics of apoptosis were comparable. Microglial numbers and responses were severely reduced...... in bone marrow chimeric mice. These results broaden our understanding of the microglial response to neural damage by demonstrating that simultaneously occurring mitosis and apoptosis regulate expansion and reduction of both resident and immigrant microglial cell populations....

  6. Modernización ecológica y políticas ambientales: notas para un debate (Brasil).

    OpenAIRE

    Olivier, Alejandro Gabriel; Domínguez, Carlos Federico

    2011-01-01

    El propósito del presente artículo es explorar conceptualmente la teoría de la modernización ecológica. Se trata de demostrar que dicha teoría, inicialmente sustentada por autores tales como Joseph Huber y Marteen Jänicke, y con desarrollos conceptuales más complejos a partir de los trabajos de Arthur Mol y Gert Spaargarem, entre otros, puede conducir a una revisión sustancial en la percepción, en la formulación y en la implementación de las políticas ambientales. El artículo concluye con el ...

  7. Note e recensioni

    Directory of Open Access Journals (Sweden)

    Autori Vari

    2012-05-01

    Full Text Available Volumi Kendall L. Walton, Mimesi come far finta [Chiara Bisignano] • Nick Zangwill, La metafisica della bellezza [Filippo Focosi] • Dominic McIver Lopes, A Philosophy of Computer Art [Elisa Caldarola] • Jacques Rancière, Béla Tarr, le temps d’après [Domenico Spinosa] • Stefano Marino, Un intreccio dialettico. Teoresi, estetica, etica e metafisica in Theo­dor W. Adorno [Marco Jacobsson] • Antonio Somaini, Ejzenštejn. Il cinema, le arti, il montaggio [Marie Rebecchi] • Aa.Vv., Alla fine delle cose. Contributi a una storia critica delle immagini [Marie Rebecchi]Convegni Merleau-Ponty et l’esthétique aujourd-hui, Università degli Studi di Milano, 5-6 maggio 2011 [Pietro Conte

  8. Metallothionein 1+2 protect the CNS during neuroglial degeneration induced by 6-aminonicotinamide

    DEFF Research Database (Denmark)

    Penkowa, Milena; Giralt, Mercedes; Camats, Jordi

    2002-01-01

    6-Aminonicotinamide (6-AN) is a niacin antagonist, which leads to degeneration of gray matter astrocytes. Metallothionein 1+2 (MT-1+2) are neuroprotective factors in the central nervous system (CNS), and to determine the roles for MT after 6-AN, we have examined transgenic mice overexpressing MT-1...... (NITT), and the number of terminal deoxynucleotidyl transferase [TdT]-mediated deoxyuridine triphosphate [dUTP]-digoxigenin nick end labeling-positive (TUNEL+), caspase-3+ apoptotic cells were significantly increased in the brainstem of normal mice after 6-AN. In the TgMTI* mice, the 6-AN-induced tissue...... damage was decreased in comparison to control mice, and they showed significantly reduced numbers of recruited macrophages and T lymphocytes, and a drastic reduction of oxidative stress and apoptotic cell death. In addition, the accompanying reactive astrogliosis was increased in the transgenic mice...

  9. Menadione-induced DNA fragmentation without 8-oxo-2'-deoxyguanosine formation in isolated rat hepatocytes

    DEFF Research Database (Denmark)

    Fischer-Nielsen, A; Corcoran, G B; Poulsen, H E

    1995-01-01

    Menadione (2-methyl-1,4-naphthoquinone) induces oxidative stress in cells causing perturbations in the cytoplasm as well as nicking of DNA. The mechanisms by which DNA damage occurs are still unclear, but a widely discussed issue is whether menadione-generated reactive oxygen species (ROS) directly...... damage DNA. In the present study, we measured the effect of menadione on formation of 7,8-dihydro-8-oxo-2'-deoxyguanosine (8-oxodG), an index of oxidative DNA base modifications, and on DNA fragmentation. Isolated hepatocytes from phenobarbital-pretreated rats were exposed to menadione, 25-400 micro......M, for 15, 90 or 180 min with or without prior depletion of reduced glutathione (GSH) by diethyl maleate. Menadione caused profound GSH depletion and internucleosomal DNA fragmentation, which was demonstrated by a prominent fragmentation ladder on agarose gel electrophoresis. We found no oxidative...

  10. Primary maths anyone can feed skittles to sharks

    CERN Document Server

    Tiley-Nunn, Nick

    2014-01-01

    Primary maths is stereotypically loved by a few hairy oddballs, tolerated by most sane primary practitioners; loathed by many. With the right approach, however; the right mindset and sense of the impossible being achievable, maths can be moulded into the diamond in the rough of the primary curriculum. Enter Nick Tiley-Nunn: Britain's most imaginative, most exciting primary maths specialist. Over years of practice he has generated ideas about the teaching of maths that are so distinct, so far out and so utterly brilliant that any primary teacher struggling to grasp the nettle of teaching long division will emerge from communing with his ideas not just with some clichéd sense that ‘maths can be fun', but that it can be brilliant, life-enhancing and truly hilarious. This book presents ideas for primary maths teaching so wildly creative and so full of the joy of life that any classroom of kids will be grateful you read it.

  11. Site directed recombination

    Science.gov (United States)

    Jurka, Jerzy W.

    1997-01-01

    Enhanced homologous recombination is obtained by employing a consensus sequence which has been found to be associated with integration of repeat sequences, such as Alu and ID. The consensus sequence or sequence having a single transition mutation determines one site of a double break which allows for high efficiency of integration at the site. By introducing single or double stranded DNA having the consensus sequence flanking region joined to a sequence of interest, one can reproducibly direct integration of the sequence of interest at one or a limited number of sites. In this way, specific sites can be identified and homologous recombination achieved at the site by employing a second flanking sequence associated with a sequence proximal to the 3'-nick.

  12. Apoptosis and microvessel density in gastric cancer: correlation with tumor stage and prognosis.

    Science.gov (United States)

    Aurello, Paolo; Bellagamba, Riccardo; Rossi Del Monte, Simone; D'Angelo, Francesco; Nigri, Giuseppe; Cicchini, Claudia; Ravaioli, Matteo; Ramacciato, Giovanni

    2009-12-01

    Gastric cancer remains one of the most common human malignancies with a poor prognosis. Apoptosis is known to be a programmed cell death and its inhibition is involved in the unregulated cellular growth that leads to neoplasms. Microvessel density (MVD) has been investigated as a promoting factor for angiogenesis with conflicting results about its relation to survival. The aim of our study was to search a correlation between these factors and some clinicopathological features and prognosis. Identification of apoptotic cells was performed applying the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling technique and recorded as apoptotic index (A.I.), whereas monoclonal antibodies were used for the study of MVD. A significant correlation was found between low and high A.I. and the subgroup of patients in Stages I and II (P stage (P = 0.036) and to a poorer 5-year overall survival (P gastric cancer.

  13. Another look at Alborz Nr. 5 in central Iran

    Energy Technology Data Exchange (ETDEWEB)

    Gretener, P.E.

    1982-04-01

    In 1956 one of the biggest blow-outs in the history of the oil industry occurred in central Iran on the Alborz structure. The well blew oil and large quantities of gas at an average rate of 60,000 bpd for 82 days for a total production in excess of 5 million bbl. The drill only nicked the reservoir, with a penetration of 2 in. Good data have been published on this event which makes this blow-out almost unique. The observations indicate that the reservoir is very highly overpressured, that the reservoir rock must have enormous permeability, and the length of the blow-out shows that rapid pressure depletion is not a problem. This indicates that Alborz is a commercial field in a high pressure environment, contrary to the widely held opinion that high pressure reservoirs are noncommercial. 10 references.

  14. Radioactivity - superstition and science

    International Nuclear Information System (INIS)

    Hinsch, Hermann

    2010-01-01

    Fairy-tales, myths, superstition - how was it fair, when we could still be afraid for witches and goblins. Where demons floated and nicks danced, the dry science has spreaded and disenchanted the life. If there would not be things like radioactivity, against which can be struggled in the collective well being. Then it is bad, clear, or good, it heals sicks, also clear. But what is now correct? In his usual humorous way the author, Dr. Hermann Hinsch, explains by means of numerous examples the phenomenon ''radioactivity'' and its effects on life. Provocantly but illustratively he illuminates, which position radioactive radiation has in our life and how and where we have already met it wantedly or unwantedly. Perhaps we must then something less shudder, but something more realism at such theme is surely not harmful.

  15. Student Travel to Pan-Am Congress of Plants & Biofuels in Merida, Mexico

    Energy Technology Data Exchange (ETDEWEB)

    Kimberly, Kimnach [American Society of Plant Biologists, Rockville, MD (United States)

    2014-04-01

    The Pan American Congress on Plants and BioEnergy convened in Mérida, Mexico, June 22 to 25, 2008. The program was organized by Steve Long (University of Illinois) and Nick Carpita (Purdue University), along with co-organizers Marcos Buckeridge (University of São Paulo, Brazil) and Federico Sánchez (Universidad Nacional Autónoma de México). More than 200 scientists from over a dozen nations around the world gathered to discuss key issues surrounding the development of biofuel feedstocks and to report on their research in this area. This three day conference had invited speakers surrounding developing renewable and sustainable energy resources which are typically propelled by three important drivers – security, cost and environmental impact.

  16. Sequential and Multistep Substrate Interrogation Provides the Scaffold for Specificity in Human Flap Endonuclease 1

    KAUST Repository

    Sobhy, M.; Joudeh, L.; Huang, X.; Takahashi, Masateru; Hamdan, S.

    2013-01-01

    Human flap endonuclease 1 (FEN1), one of the structure-specific 5' nucleases, is integral in replication, repair, and recombination of cellular DNA. The 5' nucleases share significant unifying features yet cleave diverse substrates at similar positions relative to 5' end junctions. Using single-molecule Förster resonance energy transfer, we find a multistep mechanism that verifies all substrate features before inducing the intermediary-DNA bending step that is believed to unify 5' nuclease mechanisms. This is achieved by coordinating threading of the 5' flap of a nick junction into the conserved capped-helical gateway, overseeing the active site, and bending by binding at the base of the junction. We propose that this sequential and multistep substrate recognition process allows different 5' nucleases to recognize different substrates and restrict the induction of DNA bending to the last common step. Such mechanisms would also ensure the protection ofDNA junctions from nonspecific bending and cleavage. 2013 The Authors.

  17. Leaf spray: direct chemical analysis of plant material and living plants by mass spectrometry.

    Science.gov (United States)

    Liu, Jiangjiang; Wang, He; Cooks, R Graham; Ouyang, Zheng

    2011-10-15

    The chemical constituents of intact plant material, including living plants, are examined by a simple spray method that provides real-time information on sugars, amino acids, fatty acids, lipids, and alkaloids. The experiment is applicable to various plant parts and is demonstrated for a wide variety of species. An electrical potential is applied to the plant and its natural sap, or an applied solvent generates an electrospray that carries endogenous chemicals into an adjacent benchtop or miniature mass spectrometer. The sharp tip needed to create a high electric field can be either natural (e.g., bean sprout) or a small nick can be cut in a leaf, fruit, bark, etc. Stress-induced changes in glucosinolates can be followed on the minute time scale in several plants, including potted vegetables. Differences in spatial distributions and the possibility of studying plant metabolism are demonstrated. © 2011 American Chemical Society

  18. La decision de Anne (2009: estudio de la creación de embriones genéticamente seleccionados para la curación de pacientes crónicos

    Directory of Open Access Journals (Sweden)

    Ester CASILLAS SAGRADO

    2016-09-01

    Full Text Available La decisión de Anne/ My sister´s keeper (2009 es una película de Nick Cassavetes que cuenta la historia de Anne Fitzgerald, una niña de 11 años que fue concebida para salvar a su hermana mayor, Karen, con Leucemia Promielocítica Aguda. Cuando se le plantea la necesidad de realizar un trasplante de riñón a su hermana enferma, Anne decide demandar a sus padres sobre los derechos de su cuerpo para no tener que someterse a ese procedimiento que puede condicionar para siempre su calidad de vida. Ante esta situación la madre de Anne comienza una lucha judicial contra la menor de sus hijas en un intento de disuadirla para que realice la donación aunque esto implique seguir siendo el soporte del que dependa la supervivencia de su hermana de forma indefinida.

  19. Cloning and characterization of BKV(MM) DNA and its use for detection of BKV DNA in human urine

    International Nuclear Information System (INIS)

    Harley, E.H.; Olliver, C.L.; Rhodes-Harrison, L.; Mew, R.T.; Lecatsas, G.; Naude, W. du T.

    1982-01-01

    The two fragments produced by restriction of BKV(MM) DNA with the endonucleases Pst I and Eco RI have been cloned separately into the vector pBR322 and amplified in E. coli HB101. Eight recombinant plasmids were characterized by gel electrophoresis of Pst I/Eco RI double digestions or Hind III digestions of the DNA and by hybridization of Southern gel blots to a nick-translated BKV(MM) DNA probe. Four of the recombinant plasmids contained the large Pst I/Eco RI BKV(MM) DNA fragment and four contained the small fragment. Two of these recombinant plasmids were then used to make a probe for the identification of BK DNA in a urine specimen from a patient known to be exreting particles with the morphological features of papovavirus [af

  20. Detection of human papillomavirus type 6/11 DNA in conjunctival papillomas by in situ hybridization with radioactive probes

    International Nuclear Information System (INIS)

    McDonnell, P.J.; McDonnell, J.M.; Kessis, T.; Green, W.R.; Shah, K.V.

    1987-01-01

    Twenty-three conjunctival papillomas and 28 conjunctival dysplasias were examined for human papillomavirus (HPV)-DNA sequences by in situ hybridization with nick-translated 35 S-labeled HPV probes. Adjacent paraffin sections were hybridized with HPV type 2, 6, 16, and 18 probes at Tm - 17 degrees C. Fifteen tissues, all papillomas, displayed positive hybridization with the HPV-6 probe. Infection with HPV-6 (or the closely related HPV-11) appeared to be responsible for most of the conjunctival papillomas of children and young adults. The presence of genital tract HPV-6 in these lesions suggests that some of the infections were acquired during passage through an infected birth canal. The lack of hybridization in adult conjunctival dysplasias indicates either that HPVs are not associated with this condition or that the probes and the technique utilized were not adequate for demonstration of this association

  1. Tourette's syndrome in famous musicians

    Directory of Open Access Journals (Sweden)

    Carlos Henrique F. Camargo

    2015-01-01

    Full Text Available Tourette's syndrome (TS is defined as a disorder characterized by multiple motor tics and at least one vocal tic that have lasted for not less than one year. It is a relatively complex neurobehavioral disorder, in which patients may present with coexistent attention deficit hyperactivity disorder, obsessive-compulsive disorder or other behavioral comorbidities. The musical genius Wolfgang Amadeus Mozart (1756-1791 and the rock star Kurt Cobain (1967-1994 may both have suffered from TS, and some contemporary musicians have had their clinical condition confirmed as TS. Our hypothetical diagnosis of TS in Mozart and Cobain is based on the presence of tics and psychiatric comorbidities. In contemporary musicians, such as Michael Wolff, Nick Van Bloss and James Durbin, TS has often only been diagnosed after a considerable delay. This delay in diagnosis and the controversies surrounding the clinical case of Mozart show how difficult a confirmatory diagnosis of this complex disease is.

  2. A plasmonic colorimetric strategy for visual miRNA detection based on hybridization chain reaction

    Science.gov (United States)

    Miao, Jie; Wang, Jingsheng; Guo, Jinyang; Gao, Huiguang; Han, Kun; Jiang, Chengmin; Miao, Peng

    2016-08-01

    In this work, a novel colorimetric strategy for miRNA analysis is proposed based on hybridization chain reaction (HCR)-mediated localized surface plasmon resonance (LSPR) variation of silver nanoparticles (AgNPs). miRNA in the sample to be tested is able to release HCR initiator from a solid interface to AgNPs colloid system by toehold exchange-mediated strand displacement, which then triggers the consumption of fuel strands with single-stranded tails for HCR. The final produced long nicked double-stranded DNA loses the ability to protect AgNPs from salt-induced aggregation. The stability variation of the colloid system can then be monitored by recording corresponding UV-vis spectrum and initial miRNA level is thus determined. This sensing system involves only four DNA strands which is quite simple. The practical utility is confirmed to be excellent by employing different biological samples.

  3. Study of influence of catechins on bystander responses in alpha-particle radiobiological experiments using thin PADC films

    International Nuclear Information System (INIS)

    Law, Y.L.; Yu, K.N.

    2009-01-01

    In this study, Chinese hamster ovary (CHO) cells were cultured in custom-made petri dishes with thin PADC films as substrates. Alpha particles with energies of 5 MeV were then irradiated from the bottom of PADC films. The DNA strand breaks in the bystander cells induced by irradiation were quantified with the use of terminal dUTP transferase-mediated nick end-labeling (TUNEL) assay. To study the influence of catechins on the bystander responses, catechins were added into the medium before alpha-particle irradiation of the cells. Fewer DNA strand breaks in the bystander cells were observed. As catechins are ROS (reactive oxygen species)-scavengers, the studied bystander cells might have been protected from radiation through scavenging of ROS by catechins.

  4. Sequential and Multistep Substrate Interrogation Provides the Scaffold for Specificity in Human Flap Endonuclease 1

    KAUST Repository

    Sobhy, M.

    2013-06-06

    Human flap endonuclease 1 (FEN1), one of the structure-specific 5\\' nucleases, is integral in replication, repair, and recombination of cellular DNA. The 5\\' nucleases share significant unifying features yet cleave diverse substrates at similar positions relative to 5\\' end junctions. Using single-molecule Förster resonance energy transfer, we find a multistep mechanism that verifies all substrate features before inducing the intermediary-DNA bending step that is believed to unify 5\\' nuclease mechanisms. This is achieved by coordinating threading of the 5\\' flap of a nick junction into the conserved capped-helical gateway, overseeing the active site, and bending by binding at the base of the junction. We propose that this sequential and multistep substrate recognition process allows different 5\\' nucleases to recognize different substrates and restrict the induction of DNA bending to the last common step. Such mechanisms would also ensure the protection ofDNA junctions from nonspecific bending and cleavage. 2013 The Authors.

  5. Study of influence of catechins on bystander responses in alpha-particle radiobiological experiments using thin PADC films

    Energy Technology Data Exchange (ETDEWEB)

    Law, Y.L. [Department of Physics and Materials Science, City University of Hong Kong, Tat Chee Avenue, Kowloon Tong (Hong Kong); Yu, K.N., E-mail: peter.yu@cityu.edu.h [Department of Physics and Materials Science, City University of Hong Kong, Tat Chee Avenue, Kowloon Tong (Hong Kong)

    2009-10-15

    In this study, Chinese hamster ovary (CHO) cells were cultured in custom-made petri dishes with thin PADC films as substrates. Alpha particles with energies of 5 MeV were then irradiated from the bottom of PADC films. The DNA strand breaks in the bystander cells induced by irradiation were quantified with the use of terminal dUTP transferase-mediated nick end-labeling (TUNEL) assay. To study the influence of catechins on the bystander responses, catechins were added into the medium before alpha-particle irradiation of the cells. Fewer DNA strand breaks in the bystander cells were observed. As catechins are ROS (reactive oxygen species)-scavengers, the studied bystander cells might have been protected from radiation through scavenging of ROS by catechins.

  6. Research for organism functions by analysis of radiation damage-repair process. Analysis of high order structure in radiosensitive parts

    International Nuclear Information System (INIS)

    Maekawa, Hideaki; Tsuchida, Kozo; Hashido, Kazuo; Takada, Naoko; Kameoka, Yosuke; Hirata, Makoto

    2004-01-01

    Centromere of human chromosome was recognized easily and certainly by fluorescence in situ hybridization (FISH) process. The DNA in plasmid were extracted right after irradiation of 137 Cs before repairs of the damaged DNA. Genes of the damaged DNA were detected by polymerase cycle restoration (PCR) process. Cut off frequency for two chains in the DNA were detected in real time. The cut off frequency in the damaged plasmid DNA detected by the PCR process was compared with simulation calculation. The difference between these cut off frequency values was within the value expected by electrophoretic mobility. It was cleared that the PCR amplification was difficult for the close structure of plasmid, but carried immediately on the nicked plasmid. (M. Suetake)

  7. The LHC at the AAAS

    CERN Multimedia

    CERN Bulletin

    2011-01-01

    The American Association for the Advancement of Science held its annual meeting in the Walter E. Washington Convention Center in Washington D.C. last week.   Veteran science writer Tim Radford introduces LHC scientists during a media briefing at the AAAS annual meeting. Left to right: Felicitas Pauss, Tom LeCompte, Yves Schutz and Nick Hadley. As the world’s largest popular science meeting, the AAAS meeting is a major event in the calendar of science journalists.  At this year’s LHC session, CERN’s coordinator for international relations, Felicitas Pauss, opened the discussion, paving the way for Tom LeCompte of ATLAS, Joe Incandela of CMS, Yves Schutz of ALICE and Monica Pepe-Altarelli of LHCb to report on the status of the first year’s analysis from their experiments.    

  8. Evaluation of Dying Vocal Fold Epithelial Cells by Ultrastructural Features and TUNEL Method

    Science.gov (United States)

    Novaleski, Carolyn K.; Mizuta, Masanobu; Rousseau, Bernard

    2016-01-01

    Cell death is a regulated mechanism of eliminating cells to maintain tissue homeostasis. This study described two methodological procedures for evaluating cell death in the epithelium of immobilized, approximated, and vibrated vocal folds from 12 New Zealand white breeder rabbits. The gold standard technique of transmission electron microscopy evaluated high-quality ultrastructural criteria of cell death and a common immunohistochemical marker, terminal deoxynucleotidyl transferase dUTP nick end labeling method, to confirm cell death signaling. Results revealed that ultrastructural characteristics of apoptotic cell death, specifically condensed chromatin and apoptotic bodies, were observed after vocal fold vibration and approximation. Although episodes of necrotic cell death were rare, few enlarged cell nuclei were present after vibration and approximation. The vocal fold expresses an immunohistochemical marker for apoptosis along the apical surface of the epithelium. This study provides a solid foundation for future investigations regarding the role of cell death in vocal fold health and disease. PMID:27537846

  9. Detection of programmed cell death in plant embryos.

    Science.gov (United States)

    Filonova, Lada H; Suárez, María F; Bozhkov, Peter V

    2008-01-01

    Programmed cell death (PCD) is an integral part of embryogenesis. In plant embryos, PCD functions during terminal differentiation and elimination of the temporary organ, suspensor, as well as during establishment of provascular system. Embryo abortion is another example of embryonic PCD activated at pathological situations and in polyembryonic seeds. Recent studies identified the sequence of cytological events leading to cellular self-destruction in plant embryos. As in most if not all the developmental cell deaths in plants, embryonic PCD is hallmarked by autophagic degradation of the cytoplasm and nuclear disassembly that includes breakdown of the nuclear envelope and DNA fragmentation. The optimized setup of terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) allows the routine in situ analysis of nuclear DNA fragmentation in plant embryos. This chapter provides step-by-step procedure of how to process embryos for TUNEL and how to combine TUNEL with immunolocalization of the protein of interest.

  10. Detection of Hepatitis B virus DNA and Hepatitis δ virus RNA

    International Nuclear Information System (INIS)

    Smedile, A.; Chiaberge, E.; Brunetto, M.R.; Negro, F.; Baldi, M.; Lavarini, C.; Maran, E.

    1987-01-01

    The recent availability of DNA probes of the Hepatitis B Virus DNA (HBV-DNA) and of Hepatitis Delta Virus RNA (HDV-RNA) allows the application of nucleic acid hybridization techniques to solve a variety of clinical problems. DNA probes of HBV-DNA and HDV-RNA are labeled by nick translation using 32 P or biotinylated nucleotides and hybridized to filters containing test nucleic acids. Complementary sequences are identified and the degree of blackening of the film at autoradiography or the enzymatic staining of the filter is proportional to the amount of viral nucleic acid hybridized to the probe and present in the sample. These procedures allow rapid examination of multiple specimens and are sensitive and reproducible. Viral nucleic acids can be measured quantitatively and their quantity correlates with the infectivity of sera titered in experimentally infected animals

  11. Superintelligence paths, dangers, strategies

    CERN Document Server

    Bostrom, Nick

    2014-01-01

    Superintelligence asks the questions: What happens when machines surpass humans in general intelligence? Will artificial agents save or destroy us? Nick Bostrom lays the foundation for understanding the future of humanity and intelligent life. The human brain has some capabilities that the brains of other animals lack. It is to these distinctive capabilities that our species owes its dominant position. If machine brains surpassed human brains in general intelligence, then this new superintelligence could become extremely powerful - possibly beyond our control. As the fate of the gorillas now depends more on humans than on the species itself, so would the fate of humankind depend on the actions of the machine superintelligence. But we have one advantage: we get to make the first move. Will it be possible to construct a seed Artificial Intelligence, to engineer initial conditions so as to make an intelligence explosion survivable? How could one achieve a controlled detonation? This profoundly ambitious and o...

  12. Effects of ionizing radiation on Capsicum baccatum var. pendulum (Solanaceae)

    International Nuclear Information System (INIS)

    Scaldaferro, M.A.; Prina, A.R.; Moscone, E.A.; Kwasniewska, J.

    2013-01-01

    Cytogenetic and somatic effects of various x-ray treatments were evaluated in pepper, Capsicum baccatum var. pendulum cv. “Cayenne”, with the aim to assess optimal conditions for obtaining viable lines. The cytogenetic effects were quantified by counting chromosome aberrations. The level of DNA fragmentation was estimated with TUNEL test (terminal transferase mediated dUTP-fluorescein nick end labeling). Irradiation to 20 Gy with 16-h presoaking can be a suitable treatment of the selected pepper cultivar for a mutagenesis program. - Highlights: • Cytogenetic and somatic effects of x-rays treatments in Capsicum were evaluated. • Frequencies of chromosome aberrations correlated with radiation doses. • Highest frequency of chromosome aberrations occurred with 20 Gy+soaking seeds. • In TUNEL test, the nuclei with DNA fragmentation were higher than in the control. • The strongest effects were observed with doses of 300 Gy or 20 Gy after soaking

  13. 2015 European School of High-Energy Physics

    CERN Multimedia

    2015-01-01

    Dear colleagues, I would like to draw your attention to the 2015 European School of High-Energy Physics. Details can be found at:    http://physicschool.web.cern.ch/PhysicSchool/ESHEP/ESHEP2015/default.html   The School will be held in Bulgaria from 2-15 September 2015. PLEASE NOTE THAT THE DEADLINE FOR APPLICATIONS IS 8 May 2015 The lectures will cover a broad range of HEP topics at a level suitable for students working towards a PhD in experimental particle physics.  Note that, as indicated on the website, one or two students from developing countries may be considered for the award of financial support.   Nick Ellis (On behalf of the Organising Committee)

  14. 2015 European School of High-Energy Physics

    CERN Multimedia

    2015-01-01

    Dear colleagues, I would like to draw your attention to the 2015 European School of High-Energy Physics. Details can be found at: http://physicschool.web.cern.ch/PhysicSchool/ESHEP/ESHEP2015/default.html The School will be held in Bulgaria from 2-15 September 2015. PLEASE NOTE THAT THE DEADLINE FOR APPLICATIONS IS 8 May 2015. The lectures will cover a broad range of HEP topics at a level suitable for students working towards a PhD in experimental particle physics. Note that, as indicated on the website, one or two students from developing countries may be considered for the award of financial support. Nick Ellis (On behalf of the Organising Committee)

  15. Effect of ionizing radiation on apoptosis in mouse Peyer's patches

    International Nuclear Information System (INIS)

    Liu Jiamei; Chen Dong; Liu Shuzheng

    1999-01-01

    The relationship of time-effect and dose-effect of apoptosis in mouse Peyer's patches after whole body irradiation (WBI) with different doses of X-rays was studied by the method of TdT-mediated dUTP nick end labelling (TUNEL). The results showed that the number of TUNEL positive cells in mouse Peyer's patches were significantly increased following WBI with 2 Gy irradiation, While the number of TUNEL positive cells were decreased after WBI with doses of 0.05 Gy and 0.075 Gy X-rays. the results support the view that 2 Gy irradiation promote the apoptosis of immune cells and the low doses of radiation suppress the apoptosis of immune cells

  16. Methods for decoding Cas9 protospacer adjacent motif (PAM) sequences: A brief overview.

    Science.gov (United States)

    Karvelis, Tautvydas; Gasiunas, Giedrius; Siksnys, Virginijus

    2017-05-15

    Recently the Cas9, an RNA guided DNA endonuclease, emerged as a powerful tool for targeted genome manipulations. Cas9 protein can be reprogrammed to cleave, bind or nick any DNA target by simply changing crRNA sequence, however a short nucleotide sequence, termed PAM, is required to initiate crRNA hybridization to the DNA target. PAM sequence is recognized by Cas9 protein and must be determined experimentally for each Cas9 variant. Exploration of Cas9 orthologs could offer a diversity of PAM sequences and novel biochemical properties that may be beneficial for genome editing applications. Here we briefly review and compare Cas9 PAM identification assays that can be adopted for other PAM-dependent CRISPR-Cas systems. Copyright © 2017 Elsevier Inc. All rights reserved.

  17. Purple head broccoli (Brassica oleracea L. var. italica Plenck), a functional food crop for antioxidant and anticancer potential.

    Science.gov (United States)

    Chaudhary, Ashun; Choudhary, Sonika; Sharma, Upendra; Vig, Adarsh Pal; Singh, Bikram; Arora, Saroj

    2018-05-01

    Natural foods are used in many folks and household treatments and have immense potential to treat a serious complication and health benefits, in addition to the basic nutritional values. These food products improve health, delay the aging process, increase life expectancy, and possibly prevent chronic diseases. Purple head Brassica oleracea L. var. italica Plenck is one of such foods and in current studies was explored for chemical compounds at different development stages by gas chromatography-mass spectrometry. Antioxidant potential was explored employing different assays like molybdate ion reduction, DPPH, superoxide anion radical scavenging and plasmid nicking assay. Inspired by antioxidant activity results, we further explored these extracts for antiproliferative potential by morphological changes, cell cycle analysis, measurement of intracellular peroxides and mitochondrial membrane potential changes. Current study provides the scientific basis for the use of broccoli as easily affordable potent functional food.

  18. Hegel and Dessalines: Philosophy and the African Diaspora

    Directory of Open Access Journals (Sweden)

    Deborah Jenson

    2010-12-01

    Full Text Available [First paragraph] Hegel, Haiti, and Universal History. Susan Buck-Morss. Pittsburgh: University of Pittsburgh Press, 2009. xii + 164 pp. (Paper US$ 16.95 Universal Emancipation: The Haitian Revolution and the Radical Enlightenment. Nick Nesbitt. Charlottesville: University of Virginia Press, 2008. x + 261 pp. (Paper US$ 22.50 These two books have relaunched universal history – not without controversy– as a dominant trope in the fields of colonial history and postcolonial theory. They have also highlighted tensions around the application of a Hegelian philosophical genealogy to Haiti, the first self-emancipated black postcolony, the state ghettoized as “the poorest country in the Western hemisphere,” and now the embattled zone of recovery from the catastrophic earthquake of January 2010.

  19. Unpacking Unflattening: A Conversation

    Directory of Open Access Journals (Sweden)

    Peter Wilkins

    2015-08-01

    Full Text Available 'Unflattening' (2015 is the first comic published by Harvard University Press. It is the book version of Nick Sousanis’ PhD dissertation from Teachers College, Columbia University; a project that has commanded the attention of the comics scholarship community precisely because it is comics as scholarship. This is a collaborative book review in the form of a dialogue between two authors, with each of the reviewers asking the other questions about the book; it is an effort at “unflattening.” In the responses, the reviewers have (wilfully? misunderstood each other and deviated from the question as they pursue their lines of thought. 'Unflattening' is provocative, and critical comments in the review are a result of Sousanis making us think and question. The reviewers hope that this project is not just a one-off, and that Sousanis and others continue to explore thinking through the multimodal medium of comics.

  20. Hybrid male sterility is caused by mitochondrial DNA deletion.

    Science.gov (United States)

    Hayashida, Kenji; Kohno, Shigeru

    2009-07-01

    Although it is known that the hybrid male mouse is sterile just like any other animal's heterogametic sex, the reason why only the male germ cells are impaired has yet to be discovered. TdT-mediated dUTP nick end labeling assay using a confocal fluorescence microscope and DNA fragmentation assay of hybrid testis indicated destruction of the mitochondrial DNA (mtDNA) rather than the nuclear DNA. Previously we reported that maternal mtDNA inheritance is through selective sperm mtDNA elimination based on the sperm factor and two egg factors, and expression of these three factors was recognized in the hybrid testis. It was thereby assumed that mtDNA destruction caused by the expression of maternal mtDNA inheritance system in male germ cells is implicated in the hybrid male sterility of mice.

  1. Activation of calcium-sensing receptor accelerates apoptosis in hyperplastic parathyroid cells

    International Nuclear Information System (INIS)

    Mizobuchi, Masahide; Ogata, Hiroaki; Hatamura, Ikuji; Saji, Fumie; Koiwa, Fumihiko; Kinugasa, Eriko; Koshikawa, Shozo; Akizawa, Tadao

    2007-01-01

    Calcimimetic compounds inhibit not only parathyroid hormone (PTH) synthesis and secretion, but also parathyroid cell proliferation. The aim of this investigation is to examine the effect of the calcimimetic compound NPS R-568 (R-568) on parathyroid cell death in uremic rats. Hyperplastic parathyroid glands were obtained from uremic rats (subtotal nephrectomy and high-phosphorus diet), and incubated in the media only or the media which contained high concentration of R-568 (10 -4 M), or 10% cyclodextrin, for 6 h. R-568 treatment significantly suppressed medium PTH concentration compared with that of the other two groups. R-568 treatment not only increased the number of terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling assay-positive cells, but also induced the morphologic changes of cell death determined by light or electron microscopy. These results suggest that CaR activation by R-568 accelerates parathyroid cell death, probably through an apoptotic mechanism in uremic rats in vitro

  2. A systematic review on sperm DNA fragmentation in male factor infertility: Laboratory assessment

    Directory of Open Access Journals (Sweden)

    Manesh Kumar Panner Selvam

    2018-03-01

    Full Text Available Objective: To review sperm DNA fragmentation (SDF testing as an important sperm function test in addition to conventional semen analysis. High SDF is negatively associated with semen quality, the fertilisation process, embryo quality, and pregnancy outcome. Over recent decades, different SDF assays have been developed and reviewed extensively to assess their applicability and accuracy as advanced sperm function tests. Amongst them, the standardisation of the terminal deoxynucleotidyl transferased UTP nick-end labelling (TUNEL assay with a bench top flow cytometer in clinical practice deserves special mention with a threshold value of 16.8% to differentiate infertile men with DNA damage from fertile men. Materials and methods: A systematic literature search was performed through the PubMed, Medline, and ScienceDirect databases using the keywords ‘sperm DNA fragmentation’ and ‘laboratory assessment’. Non-English articles were excluded and studies related to humans were only included. Results: Of the 618 identified, 87 studies (original research and reviews and in addition eight book chapters meeting the selection criteria were included in this review. In all, 366 articles were rejected in the preliminary screening and a further 165 articles related to non-human subjects were excluded. Conclusion: There are pros and cons to all the available SDF assays. TUNEL is a reliable technique with greater accuracy and as an additional diagnostic test in Andrology laboratories along with basic semen analysis can predict fertility outcome, and thus direct the choice of an assisted reproductive technology procedure for infertile couples. Also, the TUNEL assay can be used as a prognostic test and results are beneficial in deciding personalised treatment for infertile men. Keywords: Sperm DNA fragmentation (SDF, Terminal deoxynucleotidyl transferased UTP nick-end labelling (TUNEL, DNA damage, Sperm DNA fragmentation (SDF assay

  3. Thrombospondin-1 expression may be implicated in liver atrophic mechanism due to obstructed portal venous flow.

    Science.gov (United States)

    Hayashi, Hiromitsu; Kuroki, Hideyuki; Higashi, Takaaki; Takeyama, Hideaki; Yokoyama, Naomi; Okabe, Hirohisa; Nitta, Hidetoshi; Beppu, Toru; Takamori, Hiroshi; Baba, Hideo

    2017-07-01

    Liver is an amazing organ that can undergo regenerative and atrophic changes inversely, depending on blood flow conditions. Although the regenerative mechanism has been extensively studied, the atrophic mechanism remains to be elucidated. To assess the molecular mechanism of liver atrophy due to reduced portal blood flow, we analyzed the gene expressions between atrophic and hypertrophic livers induced by portal vein embolization in three human liver tissues using microarray analyses. Thrombospondin (TSP)-1 is an extracellular protein and a negative regulator of liver regeneration through its activation of the transforming growth factor-β/Smad signaling pathway. TSP-1 was extracted as the most upregulated gene in atrophic liver compared to hypertrophic liver due to portal flow obstruction in human. Liver atrophic and hypertrophic changes were confirmed by HE and proliferating cell nuclear antigen staining and terminal deoxynucleotidyl transferase-mediated digoxigenin-dUTP nick-end labeling. In an in vivo model with portal ligation, TSP-1 and phosphorylated Smad2 expression were continuously induced at 6 h and thereafter in the portal ligated liver, whereas the induction was transient at 6 h in the portal non-ligated liver. Indeed, while cell proliferation represented by proliferating cell nuclear antigen expression at 48 h was induced in the portal ligated liver, the sinusoidal dilatation and hepatocyte cell death with terminal deoxynucleotidyl transferase-mediated digoxigenin-dUTP nick-end labeling was detectable at 48 h in the portal ligated liver. Obstructed portal flow induces persistent TSP-1 expression and transforming growth factor-β/Smad signal activation in atrophic liver. Thrombospondin-1 may be implicated in the liver atrophic change due to obstructed portal flow as a pro-atrophic factor. © 2016 The Japan Society of Hepatology.

  4. Implementing reverse genetics in Rosaceae: analysis of T-DNA flanking sequences of insertional mutant lines in the diploid strawberry, Fragaria vesca.

    Science.gov (United States)

    Oosumi, Teruko; Ruiz-Rojas, Juan Jairo; Veilleux, Richard E; Dickerman, Allan; Shulaev, Vladimir

    2010-09-01

    Reverse genetics is used for functional genomics research in model plants. To establish a model system for the systematic reverse genetics research in the Rosaceae family, we analyzed genomic DNA flanking the T-DNA insertions in 191 transgenic plants of the diploid strawberry, Fragaria vesca. One hundred and seventy-six T-DNA flanking sequences were amplified from the right border (RB) and 37 from the left border (LB) by thermal asymmetric interlaced PCR. Analysis of the T-DNA nick positions revealed that T-DNA was most frequently nicked at the cleavage sites. Analysis of 11 T-DNA integration sites indicated that T-DNA was integrated into the F. vesca genome by illegitimate recombination, as reported in other model plants: Arabidopsis, rice and tobacco. First, deletion of DNA was found at T-DNA integration target sites in all transgenic plants tested. Second, microsimilarities of a few base pairs between the left and/or right ends of the T-DNA and genomic sites were found in all transgenic plants tested. Finally, filler DNA was identified in four break-points. Out of 191 transgenic plants, T-DNA flanking sequences of 79 plants (41%) showed significant similarity to genes, elements or proteins of other plant species and 67 (35%) of the sequences are still unknown strawberry gene fragments. T-DNA flanking sequences of 126 plants (66%) showed homology to plant ESTs. This is the first report of T-DNA integration in a sizeable population of a rosaceous species. We have shown in this paper that T-DNA integration in strawberry is not random but directed by sequence microsimilarities in the host genome.

  5. Investigation of the relationship between venticular fibrillation duration and cardiac/neurological damage in a rabbit model of electrically induced arrhythmia.

    Science.gov (United States)

    Hu, Chun-Lin; Wei, Hong-Yan; Liu, Zi-You; Li, Xing; Liao, Xiao-Xing; Li, Yu-Jie; Zhan, Hong; Jing, Xiao-Li; Xiong, Yan; Liu, Yan-Yan; Wu, Gui-Fu

    2010-12-01

    To establish a simple, economic, and reliable alternating current (AC)-induced cardiac arrest (ACCA) model in rabbits for cardiopulmonary cerebral resuscitation research. Ventricular fibrillation was induced in 27 New Zealand rabbits by external transthoracic AC, which were randomly divided into three groups according to the duration of untreated ACCA (ACCA-3 minutes, ACCA-5 minutes, and ACCA-8 minutes). After ACCA, all animals received cardiopulmonary resuscitation for 2 minutes and subsequent defibrillation until return of spontaneous circulation (ROSC). The troponin I levels were measured at 4 hours after ROSC. Animals died spontaneously or were killed at 72 hours after ROSC. The hippocampus were removed and fixed in 3% formalin. TdT-mediated dUTP-biotin nick end labeling and Nissl stainings were performed in 10-μm thickness coronal sections. Furthermore, two rabbits (without induction of ventricular fibrillation, cardiopulmonary resuscitation, and defibrillation) served as normal control group. Mean survival times after ROSC were 48.57 hours ± 24.70 hours, 18.0 hours ± 15.13 hours, and 3.88 hours ± 2.39 hours for groups ACCA-3 minutes, ACCA-5 minutes, and ACCA-8 minutes, respectively. Survival was significantly different between ACCA-3 minutes and other two groups (p = 0.002 and p = 0.01). Neuronal necrosis and apoptosis were found in the hippocampus CA1, CA2, and CA3 areas of group ACCA-3 minutes. In contrast, neuronal necrosis and TdT-mediated dUTP-biotin nick end labeling positive cells were fewer in control animals. The rabbits in group ACCA-3 minutes had significant neuronal damage with apoptosis in hippocampus CA1, CA2, and CA3 areas at 72 hours after ROSC and survived longer than those in other groups. The model we describe may be a simple, economic, and reliable model for experimental investigation on cardiopulmonary cerebral resuscitation.

  6. Aromatic residues located close to the active center are essential for the catalytic reaction of flap endonuclease-1 from hyperthermophilic archaeon Pyrococcus horikoshii.

    Science.gov (United States)

    Matsui, Eriko; Abe, Junko; Yokoyama, Hideshi; Matsui, Ikuo

    2004-04-16

    Flap endonuclease-1 (FEN-1) possessing 5'-flap endonuclease and 5'-->3' exonuclease activity plays important roles in DNA replication and repair. In this study, the kinetic parameters of mutants at highly conserved aromatic residues, Tyr33, Phe35, Phe79, and Phe278-Phe279, in the vicinity of the catalytic centers of FEN-1 were examined. The substitution of these aromatic residues with alanine led to a large reduction in kcat values, although these mutants retained Km values similar to that of the wild-type enzyme. Notably, the kcat of Y33A and F79A decreased 333-fold and 71-fold, respectively, compared with that of the wild-type enzyme. The aromatic residues Tyr33 and Phe79, and the aromatic cluster Phe278-Phe279 mainly contributed to the recognition of the substrates without the 3' projection of the upstream strand (the nick, 5'-recess-end, single-flap, and pseudo-Y substrates) for the both exo- and endo-activities, but played minor roles in recognizing the substrates with the 3' projection (the double flap substrate and the nick substrate with the 3' projection). The replacement of Tyr33, Phe79, and Phe278-Phe279, with non-charged aromatic residues, but not with aliphatic hydrophobic residues, recovered the kcat values almost fully for the substrates without the 3' projection of the upstream strand, suggesting that the aromatic groups of Tyr33, Phe79, and Phe278-Phe279 might be involved in the catalytic reaction, probably via multiple stacking interactions with nucleotide bases. The stacking interactions of Tyr33 and Phe79 might play important roles in fixing the template strand and the downstream strand, respectively, in close proximity to the active center to achieve the productive transient state leading to the hydrolysis.

  7. DNA Topology and the Initiation of Virus DNA Packaging.

    Directory of Open Access Journals (Sweden)

    Choon Seok Oh

    Full Text Available During progeny assembly, viruses selectively package virion genomes from a nucleic acid pool that includes host nucleic acids. For large dsDNA viruses, including tailed bacteriophages and herpesviruses, immature viral DNA is recognized and translocated into a preformed icosahedral shell, the prohead. Recognition involves specific interactions between the viral packaging enzyme, terminase, and viral DNA recognition sites. Generally, viral DNA is recognized by terminase's small subunit (TerS. The large terminase subunit (TerL contains translocation ATPase and endonuclease domains. In phage lambda, TerS binds a sequence repeated three times in cosB, the recognition site. TerS binding to cosB positions TerL to cut the concatemeric DNA at the adjacent nicking site, cosN. TerL introduces staggered nicks in cosN, generating twelve bp cohesive ends. Terminase separates the cohesive ends and remains bound to the cosB-containing end, in a nucleoprotein structure called Complex I. Complex I docks on the prohead's portal vertex and translocation ensues. DNA topology plays a role in the TerSλ-cosBλ interaction. Here we show that a site, I2, located between cosN and cosB, is critically important for an early DNA packaging step. I2 contains a complex static bend. I2 mutations block DNA packaging. I2 mutant DNA is cut by terminase at cosN in vitro, but in vivo, no cos cleavage is detected, nor is there evidence for Complex I. Models for what packaging step might be blocked by I2 mutations are presented.

  8. The mismatch repair and meiotic recombination endonuclease Mlh1-Mlh3 is activated by polymer formation and can cleave DNA substrates in trans.

    Science.gov (United States)

    Manhart, Carol M; Ni, Xiaodan; White, Martin A; Ortega, Joaquin; Surtees, Jennifer A; Alani, Eric

    2017-04-01

    Crossing over between homologs is initiated in meiotic prophase by the formation of DNA double-strand breaks that occur throughout the genome. In the major interference-responsive crossover pathway in baker's yeast, these breaks are resected to form 3' single-strand tails that participate in a homology search, ultimately forming double Holliday junctions (dHJs) that primarily include both homologs. These dHJs are resolved by endonuclease activity to form exclusively crossovers, which are critical for proper homolog segregation in Meiosis I. Recent genetic, biochemical, and molecular studies in yeast are consistent with the hypothesis of Mlh1-Mlh3 DNA mismatch repair complex acting as the major endonuclease activity that resolves dHJs into crossovers. However, the mechanism by which the Mlh1-Mlh3 endonuclease is activated is unknown. Here, we provide evidence that Mlh1-Mlh3 does not behave like a structure-specific endonuclease but forms polymers required to generate nicks in DNA. This conclusion is supported by DNA binding studies performed with different-sized substrates that contain or lack polymerization barriers and endonuclease assays performed with varying ratios of endonuclease-deficient and endonuclease-proficient Mlh1-Mlh3. In addition, Mlh1-Mlh3 can generate religatable double-strand breaks and form an active nucleoprotein complex that can nick DNA substrates in trans. Together these observations argue that Mlh1-Mlh3 may not act like a canonical, RuvC-like Holliday junction resolvase and support a novel model in which Mlh1-Mlh3 is loaded onto DNA to form an activated polymer that cleaves DNA.

  9. Rolling replication of UV-irradiated duplex DNA in the phi X174 replicative-form----single-strand replication system in vitro

    International Nuclear Information System (INIS)

    Shavitt, O.; Livneh, Z.

    1989-01-01

    Cloning of the phi X174 viral origin of replication into phage M13mp8 produced an M13-phi X174 chimera, the DNA of which directed efficient replicative-form----single-strand rolling replication in vitro. This replication assay was performed with purified phi X174-encoded gene A protein, Escherichia coli rep helicase, single-stranded DNA-binding protein, and DNA polymerase III holoenzyme. The nicking of replicative-form I (RFI) DNA by gene A protein was essentially unaffected by the presence of UV lesions in the DNA. However, unwinding of UV-irradiated DNA by the rep helicase was inhibited twofold as compared with unwinding of the unirradiated substrate. UV irradiation of the substrate DNA caused a strong inhibition in its ability to direct DNA synthesis. However, even DNA preparations that contained as many as 10 photodimers per molecule still supported the synthesis of progeny full-length single-stranded DNA. The appearance of full-length radiolabeled products implied at least two full rounds of replication, since the first round released the unlabeled plus viral strand of the duplex DNA. Pretreatment of the UV-irradiated DNA substrate with purified pyrimidine dimer endonuclease from Micrococcus luteus, which converted photodimer-containing supercoiled RFI DNA into relaxed, nicked RFII DNA and thus prevented its replication, reduced DNA synthesis by 70%. Analysis of radiolabeled replication products by agarose gel electrophoresis followed by autoradiography revealed that this decrease was due to a reduction in the synthesis of progeny full-length single-stranded DNA. This implies that 70 to 80% of the full-length DNA products produced in this system were synthesized on molecules that carried photodimers

  10. Discovery of a new family of relaxases in Firmicutes bacteria.

    Science.gov (United States)

    Ramachandran, Gayetri; Miguel-Arribas, Andrés; Abia, David; Singh, Praveen K; Crespo, Isidro; Gago-Córdoba, César; Hao, Jian An; Luque-Ortega, Juan Roman; Alfonso, Carlos; Wu, Ling J; Boer, D Roeland; Meijer, Wilfried J J

    2017-02-01

    Antibiotic resistance is a serious global problem. Antibiotic resistance genes (ARG), which are widespread in environmental bacteria, can be transferred to pathogenic bacteria via horizontal gene transfer (HGT). Gut microbiomes are especially apt for the emergence and dissemination of ARG. Conjugation is the HGT route that is predominantly responsible for the spread of ARG. Little is known about conjugative elements of Gram-positive bacteria, including those of the phylum Firmicutes, which are abundantly present in gut microbiomes. A critical step in the conjugation process is the relaxase-mediated site- and strand-specific nick in the oriT region of the conjugative element. This generates a single-stranded DNA molecule that is transferred from the donor to the recipient cell via a connecting channel. Here we identified and characterized the relaxosome components oriT and the relaxase of the conjugative plasmid pLS20 of the Firmicute Bacillus subtilis. We show that the relaxase gene, named relLS20, is essential for conjugation, that it can function in trans and provide evidence that Tyr26 constitutes the active site residue. In vivo and in vitro analyses revealed that the oriT is located far upstream of the relaxase gene and that the nick site within oriT is located on the template strand of the conjugation genes. Surprisingly, the RelLS20 shows very limited similarity to known relaxases. However, more than 800 genes to which no function had been attributed so far are predicted to encode proteins showing significant similarity to RelLS20. Interestingly, these putative relaxases are encoded almost exclusively in Firmicutes bacteria. Thus, RelLS20 constitutes the prototype of a new family of relaxases. The identification of this novel relaxase family will have an important impact in different aspects of future research in the field of HGT in Gram-positive bacteria in general, and specifically in the phylum of Firmicutes, and in gut microbiome research.

  11. Hyperoxygenated hydrogen-rich solution suppresses shock- and resuscitation-induced liver injury.

    Science.gov (United States)

    Dang, Yangjie; Liu, Ting; Mei, Xiaopeng; Meng, Xiangzhong; Gou, Xingchun; Deng, Bin; Xu, Hao; Xu, Lixian

    2017-12-01

    It is not known whether simultaneous delivery of hydrogen and oxygen can reduce injury caused by hemorrhagic shock and resuscitation (HSR). This study investigated the therapeutic potential of hyperoxygenated hydrogen-rich solution (HHOS), a combined hydrogen/oxygen carrier, in a rat model of HSR-induced liver injury. Rats (n = 60) were randomly divided into 5 groups (n = 6 per group at each time point). One group underwent sham operation, and the others were subjected to severe hemorrhagic shock and then treated with lactated Ringer's solution (LRS), hydrogen-rich solution, hyperoxygenated solution, or HHOS. At 2 and 6 h after resuscitation, blood samples (n = 6) were collected from the femoral artery and serum concentrations of alanine aminotransferase and aspartate aminotransferase (AST) were measured. Rats were then sacrificed, and histopathological changes in the liver were evaluated by quantifying the percentage of apoptotic cells by caspase-3 immunohistochemistry and terminal deoxynucleotidyl transferase dUTP nick-end labeling. Inflammation was assessed by assessing malondialdehyde content and tumor necrosis factor-α, and interleukin (IL)-6 expression. Compared to lactated Ringer's solution, hydrogen-rich solution, or hyperoxygenated solution groups, serum AST and alanine aminotransferase levels and IL-6, tumor necrosis factor-α, and malondialdehyde expression in liver tissue were decreased by HHOS treatment. The number of caspase-3- and terminal deoxynucleotidyl transferase dUTP nick end labeling-positive cells was decreased (P < 0.05) by HHOS treatment, 2 and 6 h after resuscitation. HHOS has protective effects against liver injury in a rat model of HSR. Copyright © 2017 Elsevier Inc. All rights reserved.

  12. Toehold-mediated strand displacement reaction triggered isothermal DNA amplification for highly sensitive and selective fluorescent detection of single-base mutation.

    Science.gov (United States)

    Zhu, Jing; Ding, Yongshun; Liu, Xingti; Wang, Lei; Jiang, Wei

    2014-09-15

    Highly sensitive and selective detection strategy for single-base mutations is essential for risk assessment of malignancy and disease prognosis. In this work, a fluorescent detection method for single-base mutation was proposed based on high selectivity of toehold-mediated strand displacement reaction (TSDR) and powerful signal amplification capability of isothermal DNA amplification. A discrimination probe was specially designed with a stem-loop structure and an overhanging toehold domain. Hybridization between the toehold domain and the perfect matched target initiated the TSDR along with the unfolding of the discrimination probe. Subsequently, the target sequence acted as a primer to initiate the polymerization and nicking reactions, which released a great abundant of short sequences. Finally, the released strands were annealed with the reporter probe, launching another polymerization and nicking reaction to produce lots of G-quadruplex DNA, which could bind the N-methyl mesoporphyrin IX to yield an enhanced fluorescence response. However, when there was even a single base mismatch in the target DNA, the TSDR was suppressed and so subsequent isothermal DNA amplification and fluorescence response process could not occur. The proposed approach has been successfully implemented for the identification of the single-base mutant sequences in the human KRAS gene with a detection limit of 1.8 pM. Furthermore, a recovery of 90% was obtained when detecting the target sequence in spiked HeLa cells lysate, demonstrating the feasibility of this detection strategy for single-base mutations in biological samples. Copyright © 2014 Elsevier B.V. All rights reserved.

  13. Leishmanicidal activities of Artemisia annua leaf essential oil against Visceral Leishmaniasis

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    Mohammad eIslamuddin

    2014-11-01

    Full Text Available Visceral leishmaniasis (VL, the second-most dreaded parasitic disease after malaria, is currently endemic in 88 countries. Dramatic increases in the rates of infection, drug resistance and non-availability of safe vaccines have highlighted the need for identification of novel and inexpensive anti-leishmanial agents from natural sources. In this study, we showed the leishmanicidal effect of essential oil from Artemisia annua leaves (AALEO against Leishmania donovani in vitro and in vivo. AALEO was extracted by hydrodistillation and characterized by GC-MS, the most abundant compounds were found to be camphor (52.06 % followed by β-caryophyllene (10.95 %. AALEO exhibited significant leishmanicidal activity against L. donovani, with 50 % inhibitory concentration of 14.63 ± 1.49 µg ml-1 and 7.3 ± 1.85 µg ml─1, respectively, against the promastigotes and intracellular amastigotes. The effect was mediated through programmed cell death as confirmed by externalization of phosphatidylserine, DNA nicking by TdT-mediated dUTP nick-end labelling (TUNEL assay, dyskinetoplastidy, cell cycle arrest at sub-G0–G1 phase, loss of mitochondrial membrane potential and reactive oxygen species (ROS generation in promastigotes and nitric oxide (NO generation in ex vivo model. AALEO presented no cytotoxic effects against mammalian macrophages even at 200 µg ml─1. Intra-peritoneal administration of AALEO (200 mg/ kg.b.w. to infected BALB/c mice reduced the parasite burden by almost 90 % in the liver and spleen with significant reduction in weight. There was no hepato- or nephro-toxicity as demonstrated by normal levels of serum enzymes. The promising antileishmanial activity shown by camphor-rich AALEO may provide a new lead in the treatment of VL.

  14. Human CD34+ cells engineered to express membrane-bound tumor necrosis factor-related apoptosis-inducing ligand target both tumor cells and tumor vasculature.

    Science.gov (United States)

    Lavazza, Cristiana; Carlo-Stella, Carmelo; Giacomini, Arianna; Cleris, Loredana; Righi, Marco; Sia, Daniela; Di Nicola, Massimo; Magni, Michele; Longoni, Paolo; Milanesi, Marco; Francolini, Maura; Gloghini, Annunziata; Carbone, Antonino; Formelli, Franca; Gianni, Alessandro M

    2010-03-18

    Adenovirus-transduced CD34+ cells expressing membrane-bound tumor necrosis factor-related apoptosis-inducing ligand (CD34-TRAIL+ cells) exert potent antitumor activity. To further investigate the mechanism(s) of action of CD34-TRAIL+ cells, we analyzed their homing properties as well as antitumor and antivascular effects using a subcutaneous myeloma model in immunodeficient mice. After intravenous injection, transduced cells homed in the tumor peaking at 48 hours when 188 plus or minus 25 CD45+ cells per 10(5) tumor cells were detected. Inhibition experiments showed that tumor homing of CD34-TRAIL+ cells was largely mediated by vascular cell adhesion molecule-1 and stromal cell-derived factor-1. Both CD34-TRAIL+ cells and soluble (s)TRAIL significantly reduced tumor volume by 40% and 29%, respectively. Computer-aided analysis of TdT-mediated dUTP nick end-labeling-stained tumor sections demonstrated significantly greater effectiveness for CD34-TRAIL+ cells in increasing tumor cell apoptosis and necrosis over sTRAIL. Proteome array analysis indicated that CD34-TRAIL+ cells and sTRAIL activate similar apoptotic machinery. In vivo staining of tumor vasculature with sulfosuccinimidyl-6-(biotinamido) hexanoate-biotin revealed that CD34-TRAIL+ cells but not sTRAIL significantly damaged tumor vasculature, as shown by TdT-mediated dUTP nick end-labeling+ endothelial cells, appearance of hemorrhagic areas, and marked reduction of endothelial area. These results demonstrate that tumor homing of CD34-TRAIL+ cells induces early vascular disruption, resulting in hemorrhagic necrosis and tumor destruction.

  15. Evidence for alternative pathways of granulosa cell death in healthy and slightly atretic bovine antral follicles.

    Science.gov (United States)

    Van Wezel, I L; Dharmarajan, A M; Lavranos, T C; Rodgers, R J

    1999-06-01

    Granulosa cell death is an early feature of atresia; however, there are many apparent contradictions in the literature concerning the mode of granulosa cell death. We have therefore examined this process in bovine healthy and atretic antral follicles, using a variety of established techniques. Light and electron microscopic observations indicated the presence of pyknotic or shrunken nuclei in both the membrana granulosa and the antrum. In the membrana granulosa, these nuclei were frequently crescent shaped and uniformly electron dense and were approximately the same size as healthy nuclei, all of which are typical of early apoptosis. However, these nuclei were within the membranes of a healthy granulosa cell, suggesting that phagocytosis by a neighboring granulosa cell is an unusually early event in the apoptotic pathway of granulosa cells. In the membrana granulosa, pyknotic nuclei stained intensely with hematoxylin but weakly with the DNA-intercalating stain propidium iodide. A percentage of these pyknotic nuclei stained by TUNEL (terminal deoxy-UTP nick end-labeling). However, in the antrum, the pyknotic nuclei and larger globules of DNA stained intensely with both hematoxylin and propidium iodide, but were not TUNEL positive. The comet assay of cell death produced a streak tail of randomly nicked DNA, rather than the plume of low mol wt apoptotic DNA. Globules collected from fresh follicular fluid stained intensely with propidium iodide and were shown by PAGE to contain DNA, the majority of which was high mol wt. In conclusion, granulosa cells within the membrana granulosa die by apoptosis, with phagocytosis by a neighboring cell preceding any potential budding of the nucleus or cell itself. Granulosa cells near the antrum are sloughed off into the antrum, and their death has features more consistent with that of other cell types that undergo death as a result of terminal differentiation.

  16. Cascade DNA nanomachine and exponential amplification biosensing.

    Science.gov (United States)

    Xu, Jianguo; Wu, Zai-Sheng; Shen, Weiyu; Xu, Huo; Li, Hongling; Jia, Lee

    2015-11-15

    DNA is a versatile scaffold for the assembly of multifunctional nanostructures, and potential applications of various DNA nanodevices have been recently demonstrated for disease diagnosis and treatment. In the current study, a powerful cascade DNA nanomachine was developed that can execute the exponential amplification of p53 tumor suppressor gene. During the operation of the newly-proposed DNA nanomachine, dual-cyclical nucleic acid strand-displacement polymerization (dual-CNDP) was ingeniously introduced, where the target trigger is repeatedly used as the fuel molecule and the nicked fragments are dramatically accumulated. Moreover, each displaced nicked fragment is able to activate the another type of cyclical strand-displacement amplification, increasing exponentially the value of fluorescence intensity. Essentially, one target binding event can induce considerable number of subsequent reactions, and the nanodevice was called cascade DNA nanomachine. It can implement several functions, including recognition element, signaling probe, polymerization primer and template. Using the developed autonomous operation of DNA nanomachine, the p53 gene can be quantified in the wide concentration range from 0.05 to 150 nM with the detection limit of 50 pM. If taking into account the final volume of mixture, the detection limit is calculated as lower as 6.2 pM, achieving an desirable assay ability. More strikingly, the mutant gene can be easily distinguished from the wild-type one. The proof-of-concept demonstrations reported herein is expected to promote the development and application of DNA nanomachine, showing great potential value in basic biology and medical diagnosis. Copyright © 2015 Elsevier B.V. All rights reserved.

  17. Characterization of a periplasmic S1-like nuclease coded by the Mesorhizobium loti symbiosis island

    International Nuclear Information System (INIS)

    Pimkin, Maxim; Miller, C. Glenn; Blakesley, Lauryn; Oleykowski, Catherine A.; Kodali, Nagendra S.; Yeung, Anthony T.

    2006-01-01

    DNA sequences encoding hypothetical proteins homologous to S1 nuclease from Aspergillus oryzae are found in many organisms including fungi, plants, pathogenic bacteria, and eukaryotic parasites. One of these is the M1 nuclease of Mesorhizobium loti which we demonstrate herein to be an enzymatically active, soluble, and stable S1 homolog that lacks the extensive mannosyl-glycosylation found in eukaryotic S1 nuclease homologs. We have expressed the cloned M1 protein in M. loti and purified recombinant native M1 to near homogeneity and have also isolated a homogeneous M1 carboxy-terminal hexahistidine tag fusion protein. Mass spectrometry and N-terminal Edman degradation sequencing confirmed the protein identity. The enzymatic properties of the purified M1 nuclease are similar to those of S1. At acidic pH M1 is 25 times more active on single-stranded DNA than on double-stranded DNA and 3 times more active on single-stranded DNA than on single-stranded RNA. At neutral pH the RNase activity of M1 exceeds the DNase activity. M1 nicks supercoiled RF-I plasmid DNA and rapidly cuts the phosphodiester bond across from the nick in the resultant relaxed RF-II plasmid DNA. Therefore, M1 represents an active bacterial S1 homolog in spite of great sequence divergence. The biochemical characterization of M1 nuclease supports our sequence alignment that reveals the minimal 21 amino acid residues that are necessarily conserved for the structure and functions of this enzyme family. The ability of M1 to degrade RNA at neutral pH implies previously unappreciated roles of these nucleases in biological systems

  18. Analysis of cis and trans Requirements for DNA Replication at the Right-End Hairpin of the Human Bocavirus 1 Genome.

    Science.gov (United States)

    Shen, Weiran; Deng, Xuefeng; Zou, Wei; Engelhardt, John F; Yan, Ziying; Qiu, Jianming

    2016-09-01

    Parvoviruses are single-stranded DNA viruses that use the palindromic structures at the ends of the viral genome for their replication. The mechanism of parvovirus replication has been studied mostly in the dependoparvovirus adeno-associated virus 2 (AAV2) and the protoparvovirus minute virus of mice (MVM). Here, we used human bocavirus 1 (HBoV1) to understand the replication mechanism of bocaparvovirus. HBoV1 is pathogenic to humans, causing acute respiratory tract infections, especially in young children under 2 years old. By using the duplex replicative form of the HBoV1 genome in human embryonic kidney 293 (HEK293) cells, we identified the HBoV1 minimal replication origin at the right-end hairpin (OriR). Mutagenesis analyses confirmed the putative NS1 binding and nicking sites within the OriR. Of note, unlike the large nonstructural protein (Rep78/68 or NS1) of other parvoviruses, HBoV1 NS1 did not specifically bind OriR in vitro, indicating that other viral and cellular components or the oligomerization of NS1 is required for NS1 binding to the OriR. In vivo studies demonstrated that residues responsible for NS1 binding and nicking are within the origin-binding domain. Further analysis identified that the small nonstructural protein NP1 is required for HBoV1 DNA replication at OriR. NP1 and other viral nonstructural proteins (NS1 to NS4) colocalized within the viral DNA replication centers in both OriR-transfected cells and virus-infected cells, highlighting a direct involvement of NP1 in viral DNA replication at OriR. Overall, our study revealed the characteristics of HBoV1 DNA replication at OriR, suggesting novel characteristics of autonomous parvovirus DNA replication. Human bocavirus 1 (HBoV1) causes acute respiratory tract infections in young children. The duplex HBoV1 genome replicates in HEK293 cells and produces progeny virions that are infectious in well-differentiated airway epithelial cells. A recombinant AAV2 vector pseudotyped with an HBoV1

  19. Feeding different levels of vitamin E and selenium has no effect on serum immunoglobulin Y (IgY production by layers vaccinated against Escherichia coli and avian encephalomyelitis virus Alimentação com diferentes níveis de vitamina E e selênio não influencia a produção de imunoglobulina Y (IgY no soro de poedeiras leves vacinadas contra Escherichia coli e encefalomielite aviária

    Directory of Open Access Journals (Sweden)

    Giselle Kindlein

    2007-10-01

    Full Text Available The effects of vitamin E and selenium (Se supplementation on the immunity of hens vaccinated against a mixture of six swine-pathogenic Escherichia coli (EC and avian encephalomyelitis virus (AEV were studied. Antibody production (AbP was evaluated in ninety 49 to 57-week-old H&N Nick Chick hens fed diets containing 14IU Vitamin E kg-1 (basal diet, 27, 59, 111, or 111IU vitamin E kg-1 + 0.56ppm Se supplementation. At 51 wks of age, half of the hens were vaccinated against EC, and all birds were vaccinated against AEV. At 53-weeks of age, the birds received a second dose of EC vaccine. Blood samples were collected weekly and serum was analyzed by ELISA for anti-EC IgY and was expressed as optical density (OD. Vaccinated hens had higher serum OD than the non-vaccinated hens (P£0.05. Vaccinated hens fed 27 and 59IU of vitamin E/kg had a higher (POs efeitos da suplementação de vitamina E e Selênio (Se na imunidade de galinhas vacinadas contra uma mistura de 6 sorotipos patogênicos de Escherichia coli (EC e o vírus da encefalomielite aviária (VEA foram estudados. A produção de anticorpos foi avaliada em galinhas H&N Nick Chick durante a 49a e 57a semanas de vida. As aves foram alimentadas com dietas contendo 14UI de Vitamina E kg-1 (dieta basal, 27, 59, 111 e 111UI de Vitamina E kg-1 + 0,56ppm Se suplementar. Às 51 semanas de idade, metade das galinhas foi vacinada contra EC, e todas as aves foram vacinadas contra VEA. Às 53 semanas, as aves receberam a segunda vacina contra EC. Amostras de sangue foram coletadas semanalmente e o soro foi analisado por ELISA para anti-EC IgY e expresso como densidade óptica (DO. Galinhas vacinadas tiveram maior DO do que as não-vacinadas (P<0,05. Aves vacinadas que receberam 27 e 59 UI de vitamina E/kg tiveram maior DO do soro (P<0,05 do que as alimentadas com 111 UI + Se. Os antígenos utilizados mostraram não ser modelos satisfatórios para estudar a influência de micronutrientes na resposta imune de

  20. Phosphodiesterase-5 inhibitor sildenafil preconditions adult cardiac myocytes against necrosis and apoptosis. Essential role of nitric oxide signaling.

    Science.gov (United States)

    Das, Anindita; Xi, Lei; Kukreja, Rakesh C

    2005-04-01

    We investigated the effect of sildenafil in protection against necrosis or apoptosis in cardiomyocytes. Adult mouse ventricular myocytes were treated with sildenafil (1 or 10 microM) for 1 h before 40 min of simulated ischemia (SI). Necrosis was determined by trypan blue exclusion and lactate dehydrogenase release following SI alone or plus 1 or 18 h of reoxygenation (RO). Apoptosis was assessed by terminal deoxynucleotidyl transferase-mediated nick end labeling assay and mitochondrial membrane potential measured using a fluorescent probe 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolyl-carbocyanine iodide (JC-1). Sildenafil reduced necrosis as indicated by decrease in trypan blue-positive myocytes and leakage of lactate dehydrogenase compared with untreated cells after either SI or SI-RO. The number of terminal deoxynucleotidyl transferase-mediated nick end labeling-positive myocytes or loss of JC-1 fluorescence following SI and 18 h of RO was attenuated in the sildenafil-treated group with concomitant inhibition of caspase 3 activity. An early increase in Bcl-2 to Bax ratio with sildenafil treatment was also observed in myocytes after SI-RO. The increase of Bcl-2 expression by sildenafil was inhibited by nitric-oxide synthase (NOS) inhibitor, L-nitro-amino-methyl-ester. The drug also enhanced mRNA and protein content of inducible NOS (iNOS) and endothelial NOS (eNOS) in the myocytes. Sildenafil-induced protection against necrosis and apoptosis was absent in the myocytes derived from iNOS knock-out mice and was attenuated in eNOS knock-out myocytes. The up-regulation of Bcl-2 expression by sildenafil was also absent in iNOS-deficient myocytes. Reverse transcription-PCR, Western blots, and immunohistochemical assay confirmed the expression of phosphodiesterase-5 in mouse cardiomyocytes. These data provide strong evidence for a direct protective effect of sildenafil against necrosis and apoptosis through NO signaling pathway. The results may have possible

  1. The synthetic inhibitor of Fibroblast Growth Factor Receptor PD166866 controls negatively the growth of tumor cells in culture

    Directory of Open Access Journals (Sweden)

    Castelli Mauro

    2009-12-01

    Full Text Available Abstract Background Many experimental data evidence that over-expression of various growth factors cause disorders in cell proliferation. The role of the Fibroblast Growth Factors (FGF in growth control is indisputable: in particular, FGF1 and its tyrosine kinase receptor (FGFR1 act through a very complex network of mechanisms and pathways. In this work we have evaluated the antiproliferative activity effect of PD166866, a synthetic molecule inhibiting the tyrosin kinase action of FGFR1. Methods Cells were routinely grown in Dulbecco Modified Eagle's medium supplemented with newborn serum and a penicillin-streptomycin mixture. Cell viability was evaluated by Mosmann assay and by trypan blue staining. DNA damage was assessed by in situ fluorescent staining with Terminal Deoxynucleotidyl Transferase dUTP nick end labeling (TUNEL assay. Assessment of oxidative stress at membrane level was measured by quantitative analysis of the intra-cellular formation of malonyl-dialdheyde (MDA deriving from the decomposition of poly-unsaturated fatty acids. The expression of Poly-ADP-Ribose-Polymerase (PARP, consequent to DNA fragmentation, was evidenced by immuno-histochemistry utilizing an antibody directed against an N-terminal fragment of the enzyme. Results The bioactivity of the drug was investigated on Hela cells. Cytoxicity was assessed by the Mosmann assay and by vital staining with trypan blue. The target of the molecule is most likely the cell membrane as shown by the significant increase of the intracellular concentration of malonyl-dihaldheyde. The increase of this compound, as a consequence of the treatment with PD166866, is suggestive of membrane lipoperoxidation. The TUNEL assay gave a qualitative, though clear, indication of DNA damage. Furthermore we demonstrate intracellular accumulation of poly-ADP-ribose polymerase I. This enzyme is a sensor of nicks on the DNA strands and this supports the idea that treatment with the drug induces cell

  2. Royal Australian and New Zealand College of Psychiatrists clinical practice guidelines for mood disorders.

    Science.gov (United States)

    Malhi, Gin S; Bassett, Darryl; Boyce, Philip; Bryant, Richard; Fitzgerald, Paul B; Fritz, Kristina; Hopwood, Malcolm; Lyndon, Bill; Mulder, Roger; Murray, Greg; Porter, Richard; Singh, Ajeet B

    2015-12-01

    , Professor John O'Brien, Professor Harold Sackeim, Professor Jan Scott, Dr Nobuhiro Sugiyama, Professor Eduard Vieta, Professor Lakshmi Yatham. Professor Marie-Paule Austin, Professor Michael Berk, Dr Yulisha Byrow, Professor Helen Christensen, Dr Nick De Felice, A/Professor Seetal Dodd, A/Professor Megan Galbally, Dr Josh Geffen, Professor Philip Hazell, A/Professor David Horgan, A/Professor Felice Jacka, Professor Gordon Johnson, Professor Anthony Jorm, Dr Jon-Paul Khoo, Professor Jayashri Kulkarni, Dr Cameron Lacey, Dr Noeline Latt, Professor Florence Levy, A/Professor Andrew Lewis, Professor Colleen Loo, Dr Thomas Mayze, Dr Linton Meagher, Professor Philip Mitchell, Professor Daniel O'Connor, Dr Nick O'Connor, Dr Tim Outhred, Dr Mark Rowe, Dr Narelle Shadbolt, Dr Martien Snellen, Professor John Tiller, Dr Bill Watkins, Dr Raymond Wu. © The Royal Australian and New Zealand College of Psychiatrists 2015.

  3. Discovery of a new family of relaxases in Firmicutes bacteria.

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    Gayetri Ramachandran

    2017-02-01

    Full Text Available Antibiotic resistance is a serious global problem. Antibiotic resistance genes (ARG, which are widespread in environmental bacteria, can be transferred to pathogenic bacteria via horizontal gene transfer (HGT. Gut microbiomes are especially apt for the emergence and dissemination of ARG. Conjugation is the HGT route that is predominantly responsible for the spread of ARG. Little is known about conjugative elements of Gram-positive bacteria, including those of the phylum Firmicutes, which are abundantly present in gut microbiomes. A critical step in the conjugation process is the relaxase-mediated site- and strand-specific nick in the oriT region of the conjugative element. This generates a single-stranded DNA molecule that is transferred from the donor to the recipient cell via a connecting channel. Here we identified and characterized the relaxosome components oriT and the relaxase of the conjugative plasmid pLS20 of the Firmicute Bacillus subtilis. We show that the relaxase gene, named relLS20, is essential for conjugation, that it can function in trans and provide evidence that Tyr26 constitutes the active site residue. In vivo and in vitro analyses revealed that the oriT is located far upstream of the relaxase gene and that the nick site within oriT is located on the template strand of the conjugation genes. Surprisingly, the RelLS20 shows very limited similarity to known relaxases. However, more than 800 genes to which no function had been attributed so far are predicted to encode proteins showing significant similarity to RelLS20. Interestingly, these putative relaxases are encoded almost exclusively in Firmicutes bacteria. Thus, RelLS20 constitutes the prototype of a new family of relaxases. The identification of this novel relaxase family will have an important impact in different aspects of future research in the field of HGT in Gram-positive bacteria in general, and specifically in the phylum of Firmicutes, and in gut microbiome

  4. Additional risk of end-of-the-pipe geoengineering technologies

    Science.gov (United States)

    Bohle, Martin

    2014-05-01

    qualitatively from the known successes. They do not tackle the initial cause, namely the carbon-dioxide inputs that are too high. This is their additional specific risk. 'The acceptability of geoengineering will be determined as much by social, legal and political issues as by scientific and technical factors', conclude Adam Corner and Nick Pidgeon (2010) when reviewing social and ethical implications of geoengineering the climate. It is to debate in that context that most geoengineering technologies are 'end of the pipe technologies', what involves an additional specific risk. Should these technologies be part of the toolbox to tackle anthropogenic climate change? Adam Corner and Nick Pidgeon 2010, Geoengineering the climate: The social and ethical implications, Environment Vol. 52.

  5. Effect of Transient Maternal Hypotension on Apoptotic Cell Death in Foetal Rat Brain

    Directory of Open Access Journals (Sweden)

    Hamit Özyürek

    2014-03-01

    Full Text Available Background: Intrauterine perfusion insufficiency induced by transient maternal hypotension has been reported to be associated with foetal brain malformations. However, the effects of maternal hypotension on apoptotic processes in the foetal brain have not been investigated experimentally during the intrauterine period. Aims: The aim of this study was to investigate the effects of transient maternal hypotension on apoptotic cell death in the intrauterine foetal brain. Study Design: Animal experimentation. Methods: Three-month-old female Wistar albino rats were allocated into four groups (n=5 each. The impact of hypoxic/ischemic injury induced by transient maternal hypotension on the 15th day of pregnancy (late gestation in rats was investigated at 48 (H17 group or 96 hours (H19 group after the insult. Control groups underwent the same procedure except for induction of hypotension (C17 and H17 groups. Brain sections of one randomly selected foetus from each pregnant rat were histopathologically evaluated for hypoxic/ischemic injury in the metencephalon, diencephalon, and telencephalon by terminal transferase-mediated dUTP nick end labelling and active cysteine-dependent aspartate-directed protease-3 (caspase-3 positivity for cell death. Results: The number of terminal transferase-mediated dUTP nick end labelling (+ cells in all the areas examined was comparable in both hypotension and control groups. The H17 group had active caspase-3 (+ cells in the metencephalon and telencephalon, sparing diencephalon, whereas the C19 and H19 groups had active caspase-3 (+ cells in all three regions. The number of active caspase-3 (+ cells in the telencephalon in the H19 group was higher compared with the metencephalon and diencephalon and compared with H17 group (p<0.05. Conclusion: Our results suggest that prenatal hypoxic/ischemic injury triggers apoptotic mechanisms. Therefore, blockade of apoptotic pathways, considering the time pattern of the insult, may

  6. La música popular de la literatura al cine: lenguaje, discurso y significados en Alta Fidelidad y The Commitments

    Directory of Open Access Journals (Sweden)

    Viñuela Suárez, Eduardo

    2010-02-01

    Full Text Available Adapting a novel into a film is not a simple question of translating the narrative plot from one medium to another, it also deals with the cultural competence of the reader / spectator and the meanings and discourses articulated in the interpretation of the texts. Music is present in both novels and films, and so it participates in the process of creating meanings, even when it does not take part in the formal structure of the text. This article develops the analysis of popular music in The Commitments and High Fidelity, two films based on two novels where the ideology of rock music influences both characters and spaces. Songs, musicians and instruments are present in prose, dialogues, sound and image, both in the novels by Roddy Doyle and Nick Hornby, respectively, and in the films directed by Alan Parker and Stephen Frears, thus demonstrating the multimedia condition of music and its capability to interact with the narrative structure of the texts in the articulation of several discourses.

    La adaptación cinematográfica de novelas va más allá de la transposición de la narrativa de un medio de expresión a otro e implica a una serie de discursos que están presentes en la obra y que son activados por el lector-espectador en función de su competencia cultural. La música, presente en ambos medios, es capaz de vehicular mensajes que, aunque no se encuentren en la estructura narrativa de las obras, participan en la creación de significados. En este artículo abordamos un análisis formal y discursivo de la música popular en Alta fidelidad y The Commitments, dos novelas adaptadas al cine en las que la ideología de la cultura rock se manifiesta en personajes y espacios. Canciones, músicos e instrumentos aparecen en texto escrito, diálogos, imagen y sonido en las novelas de Roddy Doyle y Nick Hornby y sus respectivas adaptaciones cinematográficas de Alan Parker y Stephen Frears, demostrando la condici

  7. 6th International Workshop on New Computational Methods for Inverse Problems

    International Nuclear Information System (INIS)

    2016-01-01

    methods, learning methods, convex optimization, free discontinuity problems, metamodels, proper orthogonal decomposition, reduced models for the inversion, non-linear inverse scattering, image reconstruction and restoration, and applications (bio-medical imaging, nondestructive evaluation...). NCMIP 2016 was a one-day workshop held in May 2016 which attracted around seventy attendees. Each of the submitted papers has been reviewed by two reviewers. There have been eleven accepted papers. In addition, three international speakers were invited to present a longer talk. The workshop was supported by Institut Farman (ENS Cachan, CNRS) and endorsed by the following French research networks: GDR ISIS, GDR MIA, GDR MOA, GDR Ondes. The program committee acknowledges the following research laboratories: CMLA, LMT, LURPA and SATIE. Eric Vourc'h and Thomas Rodet Workshop co-chairs: Eric Vourc'h, SATIE laboratory, Ecole Normale Supérieure de Cachan, CNRS, France Thomas Rodet, SATIE laboratory, Ecole Normale Supérieure de Cachan, CNRS, France Technical program committee: Alexandre Baussard, ENSTA Bretagne, Lab-STICC, France Marc Bonnet, ENSTA, ParisTech, France Laure Blanc-Féraud, I3S laboratory and INRIA Nice Sophia-Antipolis, France Antonin Chambolle, CMAP, Ecole Polytechnique, CNRS, France Oliver Dorn, School of Mathematics, University of Manchester, UK Cécile Durieu, SATIE, ENS Cachan, CNRS, France Laurent Fribourg, LSV, ENS Cachan, CNRS, France Jerôme Idier, IRCCyN Laboratory, Ecole Centrale de Nantes, France Pierre-Yves Joubert, IEF, Paris-Sud University, CNRS, France Marc Lambert, Geeps Laboratory, CNRS, CentraleSupElec, Paris-Sud University, France Giacomo Oliveri, eledia research center/eledia@L2S group, University of Trento, Italy Dominique Lesselier, L2S Laboratory, CNRS, CentraleSupElec, Paris-Sud University, France Matteo Pastorino, DIBE, University of Genoa, Italy Gabriel Peyré, Ceremade laboratory, University of Paris Dauphine, France Anthony Quinn

  8. Advanced glycation end products (AGEs) and their receptor (RAGE) induce apoptosis of periodontal ligament fibroblasts

    International Nuclear Information System (INIS)

    Li, D.X.; Deng, T.Z.; Lv, J.; Ke, J.

    2014-01-01

    Diabetics have an increased prevalence of periodontitis, and diabetes is one of the causative factors of severe periodontitis. Apoptosis is thought to be involved in this pathogenic relationship. The aim of this study was to investigate apoptosis in human periodontal ligament (PDL) fibroblasts induced by advanced glycation end products (AGEs) and their receptor (RAGE). We examined the roles of apoptosis, AGEs, and RAGE during periodontitis in diabetes mellitus using cultured PDL fibroblasts that were treated by AGE-modified bovine serum albumin (AGE-BSA), bovine serum albumin (BSA) alone, or given no treatment (control). Microscopy and real-time quantitative PCR indicated that PDL fibroblasts treated with AGE-BSA were deformed and expressed higher levels of RAGE and caspase 3. Cell viability assays and flow cytometry indicated that AGE-BSA reduced cell viability (69.80±5.50%, P<0.01) and increased apoptosis (11.31±1.73%, P<0.05). Hoechst 33258 staining and terminal-deoxynucleotidyl transferase-mediated nick-end labeling revealed that AGE-BSA significantly increased apoptosis of PDL fibroblasts. The results showed that the changes in PDL fibroblasts induced by AGE-BSA may explain how AGE-RAGE participates in and exacerbates periodontium destruction

  9. Bacterial reduction in genotoxicity of Direct Red 28 dye.

    Science.gov (United States)

    Bafana, Amit; Jain, Minakshi; Agrawal, Gaurav; Chakrabarti, Tapan

    2009-03-01

    Direct Red 28 (DR28) is a benzidine-based azo dye widely used in several countries. It has also been a subject of intense research for its anti-prion activity. Like other benzidine-based azo dyes, it is also carcinogenic and toxic. However, there are very few studies addressing its detoxification. In the present study, a Bacillus velezensis strain was used for detoxification of DR28. Toxicity was checked by a battery of highly sensitive genotoxicity assays like comet assay, DNA ladder formation, terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling assay and flow cytometric Annexin V binding assay. HL-60 cell line was used as the test system. All the assays showed an initial increase in toxicity upon biodegradation due to release of mutagenic products, like benzidine and 4-aminobiphenyl, from the dye. These intermediates caused significant DNA damage and induced apoptosis in HL-60 cells. Then the culture degraded these mutagenic intermediates, due to which the toxicity was reduced gradually, finally resulting in nearly complete detoxification.

  10. Hyperphosphorylated tau in the brains of mice and monkeys with long-term administration of ketamine.

    Science.gov (United States)

    Yeung, L Y; Wai, Maria S M; Fan, Ming; Mak, Y T; Lam, W P; Li, Zhen; Lu, Gang; Yew, David T

    2010-03-15

    Ketamine, a non-competitive antagonist at the glutamatergic N-methyl-d-aspartate (NMDA) receptor, might impair memory function of the brain. Loss of memory is also a characteristic of aging and Alzheimer's disease. Hyperphosphorylation of tau is an early event in the aging process and Alzheimer's disease. Therefore, we aimed to find out whether long-term ketmaine administration is related to hyperphosphorylation of tau or not in the brains of mice and monkeys. Results showed that after 6 months' administration of ketamine, in the prefrontal and entorhinal cortical sections of mouse and monkey brains, there were significant increases of positive sites for the hyperphosphorylated tau protein as compared to the control animals receiving no ketamine administration. Furthermore, about 15% of hyperphosphorylated tau positive cells were also positively labeled by terminal dUTP nick end labeling (TUNEL) indicating there might be a relationship between hyperphosphorylation of tau and apoptosis. Therefore, the long-term ketamine toxicity might involve neurodegenerative process similar to that of aging and/or Alzheimer's disease. Copyright 2010 Elsevier Ireland Ltd. All rights reserved.

  11. Effect of dietary energy and protein on the performance, egg quality, bone mineral density, blood properties and yolk fatty acid composition of organic laying hens

    Directory of Open Access Journals (Sweden)

    Md. Rakibul Hassan

    2013-02-01

    Full Text Available An experiment was conducted to evaluate the effect of dietary metabolizable energy (ME and crude protein (CP on the performance, egg quality, blood properties, bone characteristics and yolk fatty acid composition of organic laying hens. At 23 weeks, a total of 600 Brown nick laying hens were randomly distributed into 24 outdoor pens (4 replicate pens/treatment; 25 birds/pen and were given (2750, 2775 and 2800 kcal of ME/kg and CP (16 and 17% resulting in a 3×2 factorial arrangement of organic dietary treatments. The experiment lasted 23 weeks. The performance of laying hens were not affected by the dietary treatment while the egg weight was increased with energy and CP levels in the diet (P<0.05. Serum total protein was not affected by dietary energy and protein level. Total cholesterol and triglyceride tend to reduce with the increasing amount of CP in the diet. Thereafter, bone and egg quality characteristics were numerically increased in dietary 2775 kcal of ME/kg and 16% CP treatment. On the other hand, docosahexanoic acid content in egg yolk was higher (P<0.01 in 2750 kcal of ME/kg and 17% CP treatment. As a result, the performance, blood and fatty acid composition were maximized in 2750 kcal of ME/kg and 16% CP treatment. Thus, dietary 2750-2775 kcal of ME/kg and 16% CP may enhance performance, blood and fatty acid composition of organic laying hens.

  12. Dumbbell DNA-templated CuNPs as a nano-fluorescent probe for detection of enzymes involved in ligase-mediated DNA repair.

    Science.gov (United States)

    Qing, Taiping; He, Xiaoxiao; He, Dinggeng; Ye, Xiaosheng; Shangguan, Jingfang; Liu, Jinquan; Yuan, Baoyin; Wang, Kemin

    2017-08-15

    DNA repair processes are responsible for maintaining genome stability. Ligase and polynucleotide kinase (PNK) have important roles in ligase-mediated DNA repair. The development of analytical methods to monitor these enzymes involved in DNA repair pathways is of great interest in biochemistry and biotechnology. In this work, we reported a new strategy for label-free monitoring PNK and ligase activity by using dumbbell-shaped DNA templated copper nanoparticles (CuNPs). In the presence of PNK and ligase, the dumbbell-shaped DNA probe (DP) was locked and could resist the digestion of exonucleases and then served as an efficient template for synthesizing fluorescent CuNPs. However, in the absence of ligase or PNK, the nicked DP could be digested by exonucleases and failed to template fluorescent CuNPs. Therefore, the fluorescence changes of CuNPs could be used to evaluate these enzymes activity. Under the optimal conditions, highly sensitive detection of ligase activity of about 1U/mL and PNK activity down to 0.05U/mL is achieved. To challenge the practical application capability of this strategy, the detection of analyte in dilute cells extracts was also investigated and showed similar linear relationships. In addition to ligase and PNK, this sensing strategy was also extended to the detection of phosphatase, which illustrates the versatility of this strategy. Copyright © 2017 Elsevier B.V. All rights reserved.

  13. Lower sperm DNA fragmentation after r-FSH administration in functional hypogonadotropic hypogonadism.

    Science.gov (United States)

    Ruvolo, Giovanni; Roccheri, Maria Carmela; Brucculeri, Anna Maria; Longobardi, Salvatore; Cittadini, Ettore; Bosco, Liana

    2013-04-01

    An observational clinical and molecular study was designed to evaluate the effects of the administration of recombinant human FSH on sperm DNA fragmentation in men with a non-classical form of hypogonadotropic hypogonadism and idiopathic oligoasthenoteratozoospermia. In the study were included 53 men with a non-classical form of hypogonadotropic hypogonadism and idiopathic oligoasthenoteratozoospermia. In all patients, sperm DNA fragmentation index (DFI), assessed by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate (dUTP) in situ DNA nick end-labelling (TUNEL) assay, was evaluated before starting the treatment with 150 IU of recombinant human FSH, given three times a week for at least 3 months. Patients' semen analysis and DNA fragmentation index were re-evaluated after the 3-month treatment period. After recombinant human FSH therapy, we did not find any differences in terms of sperm count, motility and morphology. The average DNA fragmentation index was significantly reduced (21.15 vs 15.2, p15 %), while no significant variation occurred in the patients with DFI values ≤ 15 %. Recombinant human FSH administration improves sperm DNA integrity in hypogonadotropic hypogonadism and idiopathic oligoasthenoteratozoospermia men with DNA fragmentation index value >15 % .

  14. Assembly of α-synuclein fibrils in nanoscale studied by peptide truncation and AFM

    International Nuclear Information System (INIS)

    Zhang Feng; Lin Xiaojing; Ji Lina; Du Haining; Tang Lin; He Jianhua; Hu Jun; Hu Hongyu

    2008-01-01

    α-Synuclein (α-Syn) fibrils are the major component of Lewy bodies that are closely associated with the pathogenesis of Parkinson's disease, but the mechanism for the fibril assembly remains poorly understood. Here we report using a combination of peptide truncation and atomic force microscopy (AFM) to elucidate the self-assembly and morphology of the α-Syn fibrils. The results show that protease K significantly slims the fibrils from the mean height of ∼6.6 to ∼4.7 nm, whereas chaotropic denaturant urea completely breaks down the fibrils into small particles. The in situ enzymatic digestion also results in thinning of the fibrils, giving rise to some nicks on the fibrils. Moreover, N- or C-terminally truncated α-Syn fragments assemble into thinner filaments with the heights depending on the peptide lengths. A nine-residue peptide corresponding to the homologous GAV-motif sequence can form very thin (∼2.2 nm) but long (>1 μm) filaments. Thus, the central sequence of α-Syn forms a fibrillar core by cross-β-structure that is flanked by two flexible termini, and the orientation of the fibril growth is perpendicular to the β-sheet structures

  15. Toward the virtual screening of potential drugs in the homology modeled NAD+ dependent DNA ligase from Mycobacterium tuberculosis.

    Science.gov (United States)

    Singh, Vijai; Somvanshi, Pallavi

    2010-02-01

    DNA ligase is an important enzyme and it plays vital role in the replication and repair; also catalyzes nick joining between adjacent bases of DNA. The NAD(+) dependent DNA ligase is selectively present in eubacteria and few viruses; but missing in humans. Homology modeling was used to generate 3-D structure of NAD(+) dependent DNA ligase (LigA) of Mycobacterium tuberculosis using the known template (PDB: 2OWO). Furthermore, the stereochemical quality and torsion angle of 3-D structure was validated. Numerous effective drugs were selected and the active amino acid residue in LigA was targeted and virtual screening through molecular docking was done. In this analysis, four drugs Chloroquine, Hydroxychloroquine, Putrienscine and Adriamycin were found more potent in inhibition of M. tuberculosis through the robust binding affinity between protein-drug interactions in comparison with the other studied drugs. A phylogenetic tree was constructed and it was observed that homology of LigA in M. tuberculosis resembled with other Mycobacterium species. The conserved active amino acids of LigA may be useful to target these drugs. These findings could be used as the starting point of a rational design of novel antibacterial drugs and its analogs.

  16. Ultrasensitive electrochemical sensing platform based on graphene wrapping SnO2 nanocorals and autonomous cascade DNA duplication strategy.

    Science.gov (United States)

    Chen, Ying-Xu; Huang, Ke-Jing; Lin, Feng; Fang, Lin-Xia

    2017-12-01

    In this work, a sensitive, universal and reusable electrochemical biosensor based on stannic oxide nanocorals-graphene hybrids (SnO 2 NCs-Gr) is developed for target DNA detection by using two kinds of DNA enzymes for signal amplification through an autonomous cascade DNA duplication strategy. A hairpin probe is designed composing of a projecting part at the 3'-end as identification sequence for target, a recognition site for nicking endonuclease, and an 18-carbon shim to stop polymerization process. The designed DNA duplication-incision-replacement process is handled by KF polymerase and endonuclease, then combining with gold nanoparticles as signal carrier for further signal amplification. In the detection system, the electrochemical-chemical-chemical procedure, which uses ferrocene methanol, tris(2-carboxyethyl)phosphine and l-ascorbic acid 2-phosphate as oxidoreduction neurogen, deoxidizer and zymolyte, separately, is applied to amplify detection signal. Benefiting from the multiple signal amplification mechanism, the proposed sensor reveals a good linear connection between the peak current and logarithm of analyte concentration in range of 0.0001-1 × 10 -11 molL -1 with a detection limit of 1.25 × 10 -17 molL -1 (S/N=3). This assay also opens one promising strategy for ultrasensitive determination of other biological molecules for bioanalysis and biomedicine diagnostics. Copyright © 2017 Elsevier B.V. All rights reserved.

  17. Isothermal amplification detection of miRNA based on the catalysis of nucleases and voltammetric characteristics of silver nanoparticles.

    Science.gov (United States)

    Xu, Jianhua; Han, Kun; Liu, Dongdong; Lin, Li; Miao, Peng

    2016-11-15

    MiRNAs are a fascinating kind of biomolecule due to their vital functions in gene regulation and potential value as biomarkers for serious diseases including cancers. Exploiting convenient and sensitive methods for miRNA expression assays is imperative. In this study, we employ an exonuclease (RecJ f ) and a nicking endonuclease (Nt.BbvCI) to catalyse isothermal reactions for the amplified detection of miRNA. The degree of cyclical enzymatic amplification depends on the initial target miRNA level, which can determine the density of DNA probes bound on the electrode surface. Since DNA probes with an amino group at the 3' end are able to locate silver nanoparticles on the electrode, which provide intense stripping responses, the sensitive quantification of miRNA can be achieved. The proposed method has a limit of detection as low as 35 aM, with remarkable specificity, which offers a new approach for investigating miRNA networks and for clinical diagnosis applications.

  18. Procrastination and cramming: how adept students ace the system.

    Science.gov (United States)

    Sommer, W G

    1990-07-01

    Clear power differentials between teacher and pupil and the assignment of well-delineated tasks within specified time constraints characterize the academic system. Most university students are supremely adept at working within that structure. Many students who outwardly adapt to the system, however, engage in an intense and private ritual that comprises five aspects: calculated procrastination, preparatory anxiety, climactic cramming, nick-of-time deadline-making, and a secret, if often uncelebrated, victory. These adept students often find it difficult to admit others into their efficient program of academic survival. Although such behaviors are adaptive for school rhythms and expectations, these students may also try to impose them onto personal relationships, including those that are psychotherapeutic. The students' tendency to transfer their longstanding habits of procrastination and cramming to the workplace after graduation is less problematic. The argument is made that school rhythms and expectations shape the workplace, not vice versa. Methods of addressing the troublesome aspects of these work styles are suggested. Rather than attempting to change existing work patterns, the therapist can identify the underlying psychodynamic drama and its attendant manifestations that are becoming problematic.

  19. Binding of Bisphenol-F, a bisphenol analogue, to calf thymus DNA by multi-spectroscopic and molecular docking studies.

    Science.gov (United States)

    Usman, Afia; Ahmad, Masood

    2017-08-01

    BPF (Bisphenol-F), a member of the bisphenol family, having a wide range of industrial applications is gradually replacing Bisphenol-A. It is a recognized endocrine disrupting chemical (EDC). EDCs have been implicated in increased incidences of breast, prostate and testis cancers besides diabetes, obesity and decreased fertility. Due to the adverse effects of EDCs on human health, attempts have been directed towards their mechanism of toxicity especially at the molecular level. Hence, to understand the mechanism at the DNA level, interaction of BPF with calf thymus DNA was studied employing multi-spectroscopic, voltammetric and molecular docking techniques. Fluorescence spectra, cyclic voltammetry (CV), circular dichroism (CD) and molecular docking studies of BPF with DNA were suggestive of minor groove binding of BPF. UV-visible absorption and fluorescence spectra suggested static quenching due to complex formation between BPF and ctDNA. Hoechst 33258 (HO) and ethidium bromide (EB) displacement studies further confirmed such mode of BPF interaction. Thermodynamic and molecular docking parameters revealed the mechanism of binding of BPF with ctDNA to be favorable and spontaneous due to negative ΔG and occurring through hydrogen bonds and van der waals interactions. BPF induced DNA cleavage under in vitro conditions by plasmid nicking assay suggested it to be genotoxic. Copyright © 2017 Elsevier Ltd. All rights reserved.

  20. The roles of family B and D DNA polymerases in Thermococcus species 9°N Okazaki fragment maturation.

    Science.gov (United States)

    Greenough, Lucia; Kelman, Zvi; Gardner, Andrew F

    2015-05-15

    During replication, Okazaki fragment maturation is a fundamental process that joins discontinuously synthesized DNA fragments into a contiguous lagging strand. Efficient maturation prevents repeat sequence expansions, small duplications, and generation of double-stranded DNA breaks. To address the components required for the process in Thermococcus, Okazaki fragment maturation was reconstituted in vitro using purified proteins from Thermococcus species 9°N or cell extracts. A dual color fluorescence assay was developed to monitor reaction substrates, intermediates, and products. DNA polymerase D (polD) was proposed to function as the replicative polymerase in Thermococcus replicating both the leading and the lagging strands. It is shown here, however, that it stops before the previous Okazaki fragments, failing to rapidly process them. Instead, Family B DNA polymerase (polB) was observed to rapidly fill the gaps left by polD and displaces the downstream Okazaki fragment to create a flap structure. This flap structure was cleaved by flap endonuclease 1 (Fen1) and the resultant nick was ligated by DNA ligase to form a mature lagging strand. The similarities to both bacterial and eukaryotic systems and evolutionary implications of archaeal Okazaki fragment maturation are discussed. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

  1. Strength analysis and optimization of writing mechanism of steel billet marking machine

    Directory of Open Access Journals (Sweden)

    Fu Min

    2017-01-01

    Full Text Available According to steel billet marking theory of plasma arc nicking, the paper designs a dual laser ranging marking machine against online marking of special steel billet and realizes multi-character marking of the end face of hot steel billet. Writing mechanism bases on the rectangular coordinates marking form, Z axis adopts cantilever structure. It completes the overall marking task utilizing the synergy of KK module in X axis, Y axis and Z axis. It makes modal analysis on the writing mechanism model established by Pro/Enginner utilizing ANSYS Workbench at the position of X1Y1Z1, and obtains the first six order modal frequency and analyzes the vibration in the writing process. Moreover, the paper analyzes the static structure of the cantilever of writing mechanism, computes its maximum stress and total deformation. To make the writing mechanism reach the target of light weight, the paper optimizes Z-axis cantilever of writing mechanism. According to the analysis, it is known that the optimized Z-axis cantilever of the writing mechanism still meets the strength and rigidity requirement and total mass declines approximately 30%.

  2. DNA-dependent protein kinase inhibits AID-induced antibody gene conversion.

    Directory of Open Access Journals (Sweden)

    Adam J L Cook

    2007-04-01

    Full Text Available Affinity maturation and class switching of antibodies requires activation-induced cytidine deaminase (AID-dependent hypermutation of Ig V(DJ rearrangements and Ig S regions, respectively, in activated B cells. AID deaminates deoxycytidine bases in Ig genes, converting them into deoxyuridines. In V(DJ regions, subsequent excision of the deaminated bases by uracil-DNA glycosylase, or by mismatch repair, leads to further point mutation or gene conversion, depending on the species. In Ig S regions, nicking at the abasic sites produced by AID and uracil-DNA glycosylases results in staggered double-strand breaks, whose repair by nonhomologous end joining mediates Ig class switching. We have tested whether nonhomologous end joining also plays a role in V(DJ hypermutation using chicken DT40 cells deficient for Ku70 or the DNA-dependent protein kinase catalytic subunit (DNA-PKcs. Inactivation of the Ku70 or DNA-PKcs genes in DT40 cells elevated the rate of AID-induced gene conversion as much as 5-fold. Furthermore, DNA-PKcs-deficiency appeared to reduce point mutation. The data provide strong evidence that double-strand DNA ends capable of recruiting the DNA-dependent protein kinase complex are important intermediates in Ig V gene conversion.

  3. Bovine viral diarrhea virus: molecular cloning of genomic RNA and its diagnostic application

    International Nuclear Information System (INIS)

    Brock, K.V.

    1987-01-01

    Molecular cloning of a field isolate of bovine viral diarrhea virus (BVDV) strain 72 RNA was done in this study. The sensitivity and specificity of cloned cDNA sequences in hybridization assays with various BVDV strains were determined. cDNA was synthesized from polyadenylated BVDV RNA templates with oligo-dT primers, reverse transcriptase, and DNA polymerase I. The newly synthesized double-stranded BVDV cDNA was C-tailed with terminal deoxytransferase and annealed into G-tailed, Pst-1-cut pUC9 plasmid. Escherichia coli was transformed with the recombinant plasmids and a library of approximately 200 BVDV specific cDNA clones varying in length from 0.5 to 2.6 kilobases were isolated. The sensitivity and specificity of hybridization between the labelled cDNA and BVDV target sequences were determined. Cloned BVDV sequences were isolated from pUC9 plasmid DNA and labelled with 32 P by nick translation. The detection limit by dot blot hybridization assay was 20 pg of purified genomic BVDV RNA. cDNA hybridization probes were specific for all strains of BVDV tested, regardless of whether they were noncytopathic and cytopathic, but did not hybridize with heterologous bovine viruses tested. Probes did not hybridize with uninfected cell culture or cellular RNA. Hybridization probes were at least as sensitive as infectivity assays in detecting homologous virus

  4. [Problems with current hypertension definition in children].

    Science.gov (United States)

    Peco-Antić, Amira

    2008-01-01

    The Task Force data have added immeasurably to our understanding of the normal distribution of blood pressure in children. However, the manner in which arterial hypertension is defined in children is not without problems. The main problem is that the current definition of hypertension in children is not based on the end-organ damage assessment, but on the blood pressure height-related percentile distribution of healthy reference population. This could be overcome by introducing the relationship of blood pressure values with sensitive markers of hypertensive sequelae (such as carotid intimal-medial thickness, left ventricle mass index, retinal arteriolar narrowing and arteriovenous nicking as well as microalbumin urinary excretion) to define better the specific blood pressure values with outcomes. Furthermore, the current definition of hypertension based on the demographic and clinical characteristics of the reference population does not consider the normal evolution of reference population, as well as its ethnic differences. In addition, being based on the single occasion measurement in quite position it does not account for the possibility of transient, stress induced elevations in blood pressure known as white coat hypertension. Therefore, incorporation ambulatory 24 h blood pressure data into the definition of arterial hypertension in children may increase the definition reliability for clinical decision - making, although for such reliability the paediatric normative ambulatory blood pressure data should be improved.

  5. Efficient genome editing by FACS enrichment of paired D10A Cas9 nickases coupled with fluorescent proteins.

    Science.gov (United States)

    Gopalappa, Ramu; Song, Myungjae; Chandrasekaran, Arun Pandian; Das, Soumyadip; Haq, Saba; Koh, Hyun Chul; Ramakrishna, Suresh

    2018-05-31

    Targeted genome editing by clustered regularly interspaced short palindromic repeats (CRISPR-Cas9) raised concerns over off-target effects. The use of double-nicking strategy using paired Cas9 nickase has been developed to minimize off-target effects. However, it was reported that the efficiency of paired nickases were comparable or lower than that of either corresponding nuclease alone. Recently, we conducted a systematic comparison of the efficiencies of several paired Cas9 with their corresponding Cas9 nucleases and showed that paired D10A Cas9 nickases are sometimes more efficient than individual nucleases for gene disruption. However, sometimes the designed paired Cas9 nickases exhibited significantly lower mutation frequencies than nucleases, hampering the generation of cells containing paired Cas9 nickase-induced mutations. Here we implemented IRES peptide-conjugation of fluorescent protein to Cas9 nickase and subjected for fluorescence-activated cell sorting. The sorted cell populations are highly enriched with cells containing paired Cas9 nickase-induced mutations, by a factor of up to 40-fold as compared with the unsorted population. Furthermore, gene-disrupted single cell clones using paired nickases followed by FACS sorting strategy were generated highly efficiently, without compromising with its low off-target effects. We envision that our fluorescent protein coupled paired nickase-mediated gene disruption, facilitating efficient and highly specific genome editing in medical research.

  6. 3-Bromopyruvate induces endoplasmic reticulum stress, overcomes autophagy and causes apoptosis in human HCC cell lines.

    Science.gov (United States)

    Ganapathy-Kanniappan, Shanmugasundaram; Geschwind, Jean-Francois H; Kunjithapatham, Rani; Buijs, Manon; Syed, Labiq H; Rao, Pramod P; Ota, Shinichi; Kwak, Byung Kook; Loffroy, Romaric; Vali, Mustafa

    2010-03-01

    Autophagy, a cellular response to stress, plays a role in resistance to chemotherapy in cancer cells. Resistance renders systemic chemotherapy generally ineffective against human hepatocellular carcinoma (HCC). Recently, we reported that the pyruvate analog 3-bromopyruvate (3-BrPA) promoted tumor cell death by targeting GAPDH. In continuance, we investigated the intracellular response of two human HCC cell lines (Hep3B and SK-Hep1) that differ in their status of key apoptotic regulators, p53 and Fas. 3-BrPA treatment induced endoplasmic reticulum (ER) stress, translation inhibition and apoptosis based on Western blot and qPCR, pulse labeling, Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay and active caspase-3 in both the cell lines. However, electron microscopy revealed that 3-BrPA treated SK-Hep1 cells underwent classical apoptotic cell death while Hep3B cells initially responded with the protective autophagy that failed to prevent eventual apoptosis. 3-BrPA treatment promotes apoptosis in human HCC cell lines, irrespective of the intracellular response.

  7. Physics Meets Philosophy at the Planck Scale

    Science.gov (United States)

    Callender, Craig; Huggett, Nick

    2001-04-01

    Preface; 1. Introduction Craig Callendar and Nick Huggett; Part I. Theories of Quantum Gravity and their Philosophical Dimensions: 2. Spacetime and the philosophical challenge of quantum gravity Jeremy Butterfield and Christopher Isham; 3. Naive quantum gravity Steven Weinstein; 4. Quantum spacetime: what do we know? Carlo Rovelli; Part II. Strings: 5. Reflections on the fate of spacetime Edward Witten; 6. A philosopher looks at string theory Robert Weingard; 7. Black holes, dumb holes, and entropy William G. Unruh; Part III. Topological Quantum Field Theory: 8. Higher-dimensional algebra and Planck scale physics John C. Baez; Part IV. Quantum Gravity and the Interpretation of General Relativity: 9. On general covariance and best matching Julian B. Barbour; 10. Pre-Socratic quantum gravity Gordon Belot and John Earman; 11. The origin of the spacetime metric: Bell's 'Lorentzian Pedagogy' and its significance in general relativity Harvey R. Brown and Oliver Pooley; Part IV. Quantum Gravity and the Interpretation of Quantum Mechanics: 12. Quantum spacetime without observers: ontological clarity and the conceptual foundations of quantum gravity Sheldon Goldstein and Stefan Teufel; 13. On gravity's role in quantum state reduction Roger Penrose; 14. Why the quantum must yield to gravity Joy Christian.

  8. Antenatal taurine reduces cerebral cell apoptosis in fetal rats with intrauterine growth restriction.

    Science.gov (United States)

    Liu, Jing; Wang, Xiaofeng; Liu, Ying; Yang, Na; Xu, Jing; Ren, Xiaotun

    2013-08-15

    From pregnancy to parturition, Sprague-Dawley rats were daily administered a low protein diet to establish a model of intrauterine growth restriction. From the 12(th) day of pregnancy, 300 mg/kg rine was daily added to food until spontaneous delivery occurred. Brain tissues from normal neonatal rats at 6 hours after delivery, neonatal rats with intrauterine growth restriction, and neonatal rats with intrauterine growth restriction undergoing taurine supplement were obtained for further experiments. The terminal deoxyribonucleotidyl transferase (TdT)-mediated biotin-16-dUTP nick-end labeling assay revealed that the number of apoptotic cells in the brain tissue of neonatal rats with intrauterine growth restriction significantly increased. Taurine supplement in pregnant rats reduced cell apoptosis in brain tissue from neonatal rats with intrauterine growth restriction. nohistochemical staining revealed that taurine supplement increased glial cell line-derived neurotrophic factor expression and decreased caspase-3 expression in the cerebral cortex of intrauterine growth-restricted fetal rats. These results indicate that taurine supplement reduces cell apoptosis through the glial cell line-derived neurotrophic factor-caspase-3 signaling pathway, resulting in a protective effect on the intrauterine growth-restricted fetal rat brain.

  9. A high ratio of apoptosis to proliferation correlates with improved survival after radiotherapy for cervical adenocarcinoma

    International Nuclear Information System (INIS)

    Sheridan, Mary T.; Cooper, Rachel A.; West, Catharine M.L.

    1999-01-01

    Purpose: A retrospective study was made of the role of apoptosis in determining radiotherapy outcome in 39 adenocarcinoma of the cervix. A comparison was also made of the detection of apoptosis by morphology and the TdT dUtp nick end-labeling (TUNEL) assay. Methods and Materials: The level of apoptosis was assessed in paraffin-embedded sections by cell morphology, the TUNEL assay, and a combination of the two. A total of 2,000 cells were counted per section, to obtain apoptotic (AI) and mitotic (MI) indices. Results: Patients with a high AI had a higher survival rate than those with a low AI, however, the difference was not significant. Using a ratio of apoptosis to proliferation indices, patients with an AI:MI > median had significantly better survival than those with AI:MI < median. This was true where the AI was quantified by morphology alone (p = 0.030) or in combination with the TUNEL assay (p = 0.008). Where the AI was quantified by a combination of morphology and TUNEL, the 5-year survival rates for women with AI:MI greater or less than the median were 81% and 25%, respectively. Conclusion: A high ratio of AI:MI in adenocarcinoma of the cervix indicates a good prognosis. A combination of the TUNEL assay and morphology provided the best discrimination between outcome groups

  10. [EFFICIENCY OF INTRODUCING CAROTENE PRODUCING STRAINS BACILLUS SP. 1.1 AND B. AMYLOLIQUEFACIENS UCM B-5113 INTO THE CHIKENS DIET].

    Science.gov (United States)

    Nechypurenko, O O; Kharhota M A; Avdeeva, L V

    2015-01-01

    It was shown the efficiency of carotene producing strains Bacillus sp. 1.1 and B. amyloliquefaciens UCM B-5113 in the diet of chickens. Also it was detected the lowering of the quantitative content of bacterial genera Enterococcus, Staphylococcus, family Enterobacteriaceae in the gut after eating by chickens cross "H&N Brown Nick" fodder with strains Bacillus sp. 1.1 and B. amyloliquefaciens UCM B-5113 alone and in composition in quantities 1 x 10(10) CFU per 1 g of feed. On the 18th day after introduction of cultures Bacillus sp. 1.1, B. amyloliquefaciens UCM B-5113 and their composition in the diet of poultry we revealed the increasing of body weight by 21.6, 7.6 and 22.0%, respectively, comparesing to controls. Also due to Bacillus sp. 1.1 it was detected the restore of intestinal villous structures, tissues of spleen, liver and heart. We found the additive effect of the composition of the investigated strains of bacteria genus Bacillus to the chickens.

  11. Trichodermin induces cell apoptosis through mitochondrial dysfunction and endoplasmic reticulum stress in human chondrosarcoma cells

    International Nuclear Information System (INIS)

    Su, Chen-Ming; Wang, Shih-Wei; Lee, Tzong-Huei; Tzeng, Wen-Pei; Hsiao, Che-Jen; Liu, Shih-Chia; Tang, Chih-Hsin

    2013-01-01

    Chondrosarcoma is the second most common primary bone tumor, and it responds poorly to both chemotherapy and radiation treatment. Nalanthamala psidii was described originally as Myxosporium in 1926. This is the first study to investigate the anti-tumor activity of trichodermin (trichothec-9-en-4-ol, 12,13-epoxy-, acetate), an endophytic fungal metabolite from N. psidii against human chondrosarcoma cells. We demonstrated that trichodermin induced cell apoptosis in human chondrosarcoma cell lines (JJ012 and SW1353 cells) instead of primary chondrocytes. In addition, trichodermin triggered endoplasmic reticulum (ER) stress protein levels of IRE1, p-PERK, GRP78, and GRP94, which were characterized by changes in cytosolic calcium levels. Furthermore, trichodermin induced the upregulation of Bax and Bid, the downregulation of Bcl-2, and the dysfunction of mitochondria, which released cytochrome c and activated caspase-3 in human chondrosarcoma. In addition, animal experiments illustrated reduced tumor volume, which led to an increased number of terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL)-positive cells and an increased level of cleaved PARP protein following trichodermin treatment. Together, this study demonstrates that trichodermin is a novel anti-tumor agent against human chondrosarcoma cells both in vitro and in vivo via mitochondrial dysfunction and ER stress. - Highlights: • Trichodermin induces chondrosarcoma apoptosis. • ER stress is involved in trichodermin-induced cell death. • Trichodermin induces chondrosarcoma death in vivo.

  12. Clustered Regularly Interspaced Short Palindromic Repeats/Cas9 Triggered Isothermal Amplification for Site-Specific Nucleic Acid Detection.

    Science.gov (United States)

    Huang, Mengqi; Zhou, Xiaoming; Wang, Huiying; Xing, Da

    2018-02-06

    A novel CRISPR/Cas9 triggered isothermal exponential amplification reaction (CAS-EXPAR) strategy based on CRISPR/Cas9 cleavage and nicking endonuclease (NEase) mediated nucleic acids amplification was developed for rapid and site-specific nucleic acid detection. CAS-EXPAR was primed by the target DNA fragment produced by cleavage of CRISPR/Cas9, and the amplification reaction performed cyclically to generate a large number of DNA replicates which were detected using a real-time fluorescence monitoring method. This strategy that combines the advantages of CRISPR/Cas9 and exponential amplification showed high specificity as well as rapid amplification kinetics. Unlike conventional nucleic acids amplification reactions, CAS-EXPAR does not require exogenous primers, which often cause target-independent amplification. Instead, primers are first generated by Cas9/sgRNA directed site-specific cleavage of target and accumulated during the reaction. It was demonstrated this strategy gave a detection limit of 0.82 amol and showed excellent specificity in discriminating single-base mismatch. Moreover, the applicability of this method to detect DNA methylation and L. monocytogenes total RNA was also verified. Therefore, CAS-EXPAR may provide a new paradigm for efficient nucleic acid amplification and hold the potential for molecular diagnostic applications.

  13. The Roles of Family B and D DNA Polymerases in Thermococcus Species 9°N Okazaki Fragment Maturation*

    Science.gov (United States)

    Greenough, Lucia; Kelman, Zvi; Gardner, Andrew F.

    2015-01-01

    During replication, Okazaki fragment maturation is a fundamental process that joins discontinuously synthesized DNA fragments into a contiguous lagging strand. Efficient maturation prevents repeat sequence expansions, small duplications, and generation of double-stranded DNA breaks. To address the components required for the process in Thermococcus, Okazaki fragment maturation was reconstituted in vitro using purified proteins from Thermococcus species 9°N or cell extracts. A dual color fluorescence assay was developed to monitor reaction substrates, intermediates, and products. DNA polymerase D (polD) was proposed to function as the replicative polymerase in Thermococcus replicating both the leading and the lagging strands. It is shown here, however, that it stops before the previous Okazaki fragments, failing to rapidly process them. Instead, Family B DNA polymerase (polB) was observed to rapidly fill the gaps left by polD and displaces the downstream Okazaki fragment to create a flap structure. This flap structure was cleaved by flap endonuclease 1 (Fen1) and the resultant nick was ligated by DNA ligase to form a mature lagging strand. The similarities to both bacterial and eukaryotic systems and evolutionary implications of archaeal Okazaki fragment maturation are discussed. PMID:25814667

  14. Reconstitution of nucleotide excision nuclease with UvrA and UvrB proteins from Escherichia coli and UvrC protein from Bacillus subtilis

    International Nuclear Information System (INIS)

    Lin, J.J.; Sancar, A.

    1990-01-01

    Recently, an open reading frame which has a deduced amino acid sequence that shows 38% homology to Escherichia coli UvrC protein was found upstream of the aspartokinase II gene (ask) in Bacillus subtilis. We found that plasmids containing this open reading frame complement the uvrC mutations in E. coli. We joined the open reading frame to a tac promoter to amplify the gene product in E. coli and purified the protein to near homogeneity. The apparent molecular weight of the gene product is 69,000, which is consistent with the calculated molecular weight of 69,378 fro the deduced gene product of the open reading frame. The purified gene product causes the nicking of DNA at the 8th phosphodiester bond 5' and the 5th phosphodiester bond 3' to a thymine dimer when mixed with E. coli UvrA and UvrB proteins and a DNA substrate containing a uniquely located thymine dimer. We conclude that the gene product of the open reading frame is the B. subtilis UvrC protein. Our results suggest that the B. subtilis nucleotide excision repair system is quite similar to that of E. coli. Furthermore, complementation of the UvrA and UvrB proteins from a Gram-negative bacterium with the UvrC protein of Gram-positive B. subtilis indicates a significant evolutionary conservation of the nucleotide excision repair system

  15. Trichodermin induces cell apoptosis through mitochondrial dysfunction and endoplasmic reticulum stress in human chondrosarcoma cells

    Energy Technology Data Exchange (ETDEWEB)

    Su, Chen-Ming [Graduate Institute of Basic Medical Science, China Medical University, Taichung, Taiwan (China); Wang, Shih-Wei [Department of Medicine, Mackay Medical College, New Taipei City, Taiwan (China); Lee, Tzong-Huei [Graduate Institute of Pharmacognosy, Taipei Medical University, Taipei, Taiwan (China); Tzeng, Wen-Pei [Graduate Institute of Sports and Health, National Changhua University of Education, Changhua, Taiwan (China); Hsiao, Che-Jen [School of Respiratory Therapy, College of Medicine, Taipei Medical University, Taipei, Taiwan (China); Liu, Shih-Chia [Department of Orthopaedics, Mackay Memorial Hospital, Taipei, Taiwan (China); Tang, Chih-Hsin, E-mail: chtang@mail.cmu.edu.tw [Graduate Institute of Basic Medical Science, China Medical University, Taichung, Taiwan (China); Department of Pharmacology, School of Medicine, China Medical University, Taichung, Taiwan (China); Department of Biotechnology, College of Health Science, Asia University, Taichung, Taiwan (China)

    2013-10-15

    Chondrosarcoma is the second most common primary bone tumor, and it responds poorly to both chemotherapy and radiation treatment. Nalanthamala psidii was described originally as Myxosporium in 1926. This is the first study to investigate the anti-tumor activity of trichodermin (trichothec-9-en-4-ol, 12,13-epoxy-, acetate), an endophytic fungal metabolite from N. psidii against human chondrosarcoma cells. We demonstrated that trichodermin induced cell apoptosis in human chondrosarcoma cell lines (JJ012 and SW1353 cells) instead of primary chondrocytes. In addition, trichodermin triggered endoplasmic reticulum (ER) stress protein levels of IRE1, p-PERK, GRP78, and GRP94, which were characterized by changes in cytosolic calcium levels. Furthermore, trichodermin induced the upregulation of Bax and Bid, the downregulation of Bcl-2, and the dysfunction of mitochondria, which released cytochrome c and activated caspase-3 in human chondrosarcoma. In addition, animal experiments illustrated reduced tumor volume, which led to an increased number of terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL)-positive cells and an increased level of cleaved PARP protein following trichodermin treatment. Together, this study demonstrates that trichodermin is a novel anti-tumor agent against human chondrosarcoma cells both in vitro and in vivo via mitochondrial dysfunction and ER stress. - Highlights: • Trichodermin induces chondrosarcoma apoptosis. • ER stress is involved in trichodermin-induced cell death. • Trichodermin induces chondrosarcoma death in vivo.

  16. Σκέψεις για δύο μεσαιωνικά τοπωνύμια της Κριμαίας (Αλούστου, Παρθενίται

    Directory of Open Access Journals (Sweden)

    Παντελής ΧΑΡΑΛΑΜΠΑΚΗΣ

    2014-02-01

    Full Text Available In the present study the author tries to examine two medieval place names of the Crimea: Aloustou (Ἀλούστου and Parthenitai (Παρθενίται. The Greek name Aloustou is in genitive and it should originally be a place name formed by a personal name or rather a nick name aloustos (ἄλουστος. In the late Middle Ages the name changed to Alusta (Ἀλούστα, thus becoming a female one. The loss of the initial a- in the name (Lusta attested in some Latin and Italian documents can be explained. The name Parthenitai (in plural is actually a place name formed by an ethnic name Parthenitai. The ethnic name derived from a place name Parthenion (Παρθένιον. In the late Middle Ages the name Parthenitai also changed to a female one, Parthenita (Παρθενίτα.

  17. The Root Extract of Gentiana macrophylla Pall. Alleviates Cardiac Apoptosis in Lupus Prone Mice.

    Directory of Open Access Journals (Sweden)

    Chih-Yang Huang

    Full Text Available The roots of the perennial herb Gentiana macrophylla Pall. (GM are known as Qinjiao, which has been used for centuries to treat systemic lupus erythematosus (SLE. However, little is known about the effects of GM on cholesterol-aggravated cardiac abnormalities in SLE, and the mechanisms thereof. This study investigates whether GM exhibits anti-apoptotic effects, focusing on the left ventricle (LV of NZB/W F1 mice fed with high-cholesterol diet. The morphology and apoptotic status of ventricular tissues were determined by microscopy and Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL assay. Levels of apoptotic biomarkers were determined by immunoblotting. The results thus obtained revealed that GM significantly reduced the cholesterol-aggravated apoptosis of LV in NZB/W F1 mice by suppressing both intrinsic and extrinsic apoptotic pathways. Additionally, GM significantly increased the cardiac insulin-like growth factors (IGF-1 survival signaling and anti-apoptotic proteins in LV tissues. Accordingly, GM is considered to be beneficial in alleviating cholesterol-aggravated cardiac damage in SLE, and therefore constitute an alternative treatment for SLE patients with cardiac abnormalities.

  18. Enhancing colour and oxidative stabilities of reduced-nitrite turkey meat sausages during refrigerated storage using fucoxanthin purified from the Tunisian seaweed Cystoseira barbata.

    Science.gov (United States)

    Sellimi, Sabrine; Ksouda, Ghada; Benslima, Abdelkarim; Nasri, Rim; Rinaudo, Marguerite; Nasri, Moncef; Hajji, Mohamed

    2017-09-01

    The present study investigated the angiotensin-I converting enzyme (ACE) inhibitory activity and the antioxidant properties, in vitro and in cured meat sausages containing reduced levels of sodium nitrite, of fucoxanthin extracted from the Tunisian brown seaweed Cystoseira barbata (CBFX). Results revealed that CBFX exhibited great scavenging activities against DPPH free radicals (EC 50  = 136 μg/ml), peroxyl radicals in the linoleate-β-carotene system (EC 50  = 43 μg/ml) and hydroxyl radicals generated by Fenton reaction (DNA nicking assay). A considerable ferric reducing potential was also recorded for CBFX (EC 50  = 34 μg/ml). It is interesting to note that CBFX was found to modulate the ACE activity, which is the key enzyme involved in the blood pressure regulation, with an EC 50 of 5 μg/ml. When fucoxanthin was supplemented, the concentration of sodium nitrite added to cured turkey meat sausages was reduced from 150 to 80 ppm, coupled with the enhancement of colour and oxidative stabilities. Thus, CBFX, with noticeable antioxidant and antihyertensive effects, could be used as a natural additive in functional foods to alleviate potential human health hazards caused by carcinogenic nitrosamines formation. Copyright © 2017 Elsevier Ltd. All rights reserved.

  19. Neuropeptide Treatment with Cerebrolysin Enhances the Survival of Grafted Neural Stem Cell in an α-Synuclein Transgenic Model of Parkinson's Disease

    Directory of Open Access Journals (Sweden)

    Edward Rockenstein

    2015-01-01

    Full Text Available Neuronal stem cell (NSC grafts have been investigated as a potential neuro-restorative therapy in Parkinson's disease (PD but their use is compromised by the death of grafted cells. We investigated the use of Cerebrolysin (CBL, a neurotrophic peptide mixture, as an adjunct to NSC therapy in the α-synuclein (α-syn transgenic (tg model of PD. In vehicle-treated α-syn tg mice, there was decreased survival of NSCs. In contrast, CBL treatment enhanced the survival of NSCs in α-syn tg groups and ameliorated behavioral deficits. The grafted NSCs showed lower levels of terminal deoxynucleotidyl transferase dUTP nick end labeling positive cells in the CBL-treated mice when compared with vehicle-treated α-syn tg mice. No evidence of tumor growth was detected. Levels of α-syn were similar in the vehicle in CBL-treated tg mice. In conclusion, CBL treatment might be a potential adjuvant for therapeutic NSC grafting in PD.

  20. In vitro replication of poliovirus

    International Nuclear Information System (INIS)

    Lubinski, J.M.

    1986-01-01

    Poliovirus is a member of the Picornaviridae whose genome is a single stranded RNA molecule of positive polarity surrounded by a proteinaceous capsid. Replication of poliovirus occurs via negative strand intermediates in infected cells using a virally encoded RNA-dependent RNA polymerase and host cell proteins. The authors have exploited the fact that complete cDNA copies of the viral genome when transfected onto susceptible cells generate virus. Utilizing the bacteriophage SP6 DNA dependent RNA polymerase system to synthesize negative strands in vitro and using these in an in vitro reaction the authors have generated full length infectious plus strands. Mutagenesis of the 5' and 3' ends of the negative and positive strands demonstrated that replication could occur either de novo or be extensions of the templates from their 3' ends or from nicks occurring during replication. The appearance of dimeric RNA molecules generated in these reactions was not dependent upon the same protein required for de novo initiation. Full length dimeric RNA molecules using a 5' 32 P end-labelled oligo uridylic acid primer and positive strand template were demonstrated in vitro containing only the 35,000 Mr host protein and the viral RNA-dependent RNA polymerase. A model for generating positive strands without protein priming by cleavage of dimeric RNA molecules was proposed

  1. Effect of erythropoietin on acoustically traumatized rat cochlea: an immunohistochemical study.

    Science.gov (United States)

    Gürgen, Oğuzhan; Gürgen, Seren Gülşen; Kirkim, Günay; Kolatan, Efsun; Gürkan, Selhan; Güvenç, Yeşim; Eskiizmir, Görkem

    2014-08-01

    To investigate the audiological and histopathological effects of erythropoietin on acoustic overstimulation in rats. Twenty-two male Wistar albino rats were divided into 3 groups: sham group (n = 7), erythropoietin injection group (n = 8), and saline injection group (n = 7). Both erythropoietin and saline injection groups were exposed to white noise (100 decibel [dB] sound pressure level [SPL]) for 3 hours. Auditory brainstem responses were measured before, immediately after, and on the 7th day of noise exposure. All animals were sacrificed on the 7th day and temporal bones were collected. The serial sections of the cochleae were stained by caspase-3 and caspase-9 immunostaining and by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) method in order to detect apoptotic cells. In the saline group statistically significant differences were detected between the baseline and immediate postacoustic overstimulation thresholds of click and 6 kHz stimuli. However, when the baseline and immediate postacoustic overstimulation thresholds of click and 6 kHz stimuli were compared in the erythropoietin injection group, no statistically significant difference was determined. Histopathologic evaluations demonstrated that erythropoietin decreased the amount of apoptotic cells in the cochlea. Erythropoietin is likely to prevent the acute threshold changes and decrease the amount of apoptosis in cochlea after acoustic overstimulation in rats.

  2. Time-specific blockade of PDGFR with Imatinib (Glivec®) causes cataract and disruption of lens fiber cells in neonatal mice.

    Science.gov (United States)

    Zhou, Yin-Pin; He, Yang-Tao; Chen, Cheng-Li; Ji, Jun; Niu, Jian-Qin; Wang, Han-Zhi; Li, Shi-Feng; Huang, Lan; Mei, Feng

    2011-03-01

    This study aimed at investigating the response of lens epithelial cells in postnatal mice to Imatinib (Glivec®, a potent inhibitor of platelet-derived growth factor receptor (PDGFR)) treatment. Mouse eyes were sampled 10 days after administration of Imatinib (0.5 mg·g(-1)·day(-1)) for 3 days, at either 7, 14, or 21 days postpartum. Structural changes of lens were revealed by routine H.E. staining. Levels of proliferation and apoptosis were revealed by BrdU incorporation and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay, respectively, and immunofluorescent staining with anti-PDGFRα antibody was carried out on the sections of eyeball. PDGFRα and p-PDGFRαprotein levels were evaluated by Western blot. Our results indicated that administration of Imatinib led to blockade of PDGFR signaling. Formation of cataracts was found only in those mice where treatment started from 7 days postpartum (P7), but was not observed in those samples from P14 nor P21. Fiber cells were disorganized in cataract lens core as observed histologically, and migration of epithelial cells was also inhibited. No apoptosis was detected with the TUNEL method. Our results indicated blockade of PDGFR at the neonatal stage (P7) would lead to cataracts and lens fiber cells disorganization, suggesting that PDGFR signaling plays a time-specific and crucial role in the postnatal development of lens in the mouse, and also may provide a new approach to produce a congenital cataract animal model.

  3. Design of a Lunar Quick-Attach Mechanism to Hummer Vehicle Mounting Interface

    Science.gov (United States)

    Grismore, David A.

    2010-01-01

    This report presents my work experiences while I was an intern with NASA (National Aeronautic and Space Administration) in the Spring of2010 at the Kennedy Space Center (KSC) launch facility in Cape Canaveral, Florida as a member of the NASA USRP (Undergraduate Student Research Program) program. I worked in the Surface Systems (NE-S) group during the internship. Within NE-S, two ASRC (Arctic Slope Regional Corporation) contract engineers, A.J. Nick and Jason Schuler, had developed a "Quick-Attach" mechanism for the Chariot Rover, the next generation lunar rover. My project was to design, analyze, and possibly fabricate a mounting interface between their "Quick-Attach" and a Hummer vehicle. This interface was needed because it would increase their capabilities to test the Quick Attach and its various attachments, as they do not have access to a Chariot Rover at KSC. I utilized both Pro Engineer, a 3D CAD software package, and a Coordinate Measuring Machine (CMM) known as a FAROarm to collect data and create my design. I relied on hand calculations and the Mechanica analysis tool within Pro Engineer to perform stress analysis on the design. After finishing the design, I began working on creating professional level CAD drawings and issuing them into the KSC design database known as DDMS before the end of the internship.

  4. Panel discussion: Building Canadian companies and capabilities in the transition to the hydrogen economy

    International Nuclear Information System (INIS)

    Beck, N.

    2004-01-01

    'Full text:' Moderated by Nick Beck from Natural Resources Canada, this panel discussion will be prefaced by a keynote address by Dr. Arthur Carty, Canada's National Science Advisor and former President of the National Research Council of Canada, who will discuss technology commercialization in Canada and how the Government of Canada and industry collaborate to achieve their respective priorities. This session will illustrate innovative government and industry partnerships from early research and development to project demonstration, and adoption into the market-place. Panelists from across the Canadian hydrogen and fuel cell innovation spectrum will provide an overview of their respective company's partnerships with the Government of Canada and speak to how these alliances have helped their company to pioneer new technology, move technology from the lab to the marketplace, and/or become more competitive. Opportunities and challenges that companies have faced in their partnerships with government will be shared with the audience. The Panel Members are: Mr. Stephen Kukucha, Mr. Chris Reid, Mr. Robb Thompson, Mr. Pierre Rivard, Mr. John Shen, Mr. R. Randall MacEwen, Mr. Jonathan Wilkinson. Companies will also be showcasing their contribution in advancing Canada's and the world's transition to the hydrogen economy. (author)

  5. Markers of potential malignancy in chronic hyperplastic candidiasis.

    Science.gov (United States)

    Darling, Mark R; McCord, Christina; Jackson-Boeters, Linda; Copete, Maria

    2012-08-01

    To examine the presence of markers associated with malignancy, including p53, p21 cyclin-dependent kinase inhibitor 1A, murine double minutes-2, and others, in chronic hyperplastic candidiasis. Immunohistochemical methods were used to examine the expression of p53, murine double minutes-2, p21 cyclin-dependent kinase inhibitor 1A, metallothionein, and proliferating cell nuclear antigen in 42 chronic hyperplastic candidiasis lesions and 11 non-infected control tissues. Terminal deoxynucleotidyl transferase-mediated digoxigenin-dUTP nick-end labeling was used to examine apoptosis, which was correlated with p53 expression. These markers were measured in lesions of chronic hyperplastic candidiasis that did not show any epithelial dysplasia or histological signs of malignancy. p53 scores were higher in chronic hyperplastic candidiasis than in controls (P = 0.0046). Murine double-minutes 2 levels were not elevated. p21 cyclin-dependent kinase inhibitor 1A was increased in parabasal (P candidiasis lesions showed a similar basal/parabasal metallothionein staining pattern to that seen in normal squamous epithelium. Proliferating cell nuclear antigen was increased (P = 0.0007), as was apoptosis (P = 0.0033). Increased p53 in oral chronic hyperplastic candidiasis suggests an increased potential for malignant change in the epithelium, above that of normal tissues. Further functional investigation is required, as well as clinical follow-up studies. © 2012 Blackwell Publishing Asia Pty Ltd.

  6. Anticancer Activity of Indian Stingless Bee Propolis: An In Vitro Study

    Directory of Open Access Journals (Sweden)

    Milind K. Choudhari

    2013-01-01

    Full Text Available Indian stingless bee propolis has a complex chemical nature and is reported to possess various medicinal properties. In the present study, anticancer activity of the ethanolic extract of propolis (EEP was explored by testing the cytotoxic and apoptotic effect in four different cancer cell lines, namely, MCF-7 (human breast cancer, HT-29 (human colon adenocarcinoma, Caco-2 (human epithelial colorectal adenocarcinoma, and B16F1 (murine melanoma, at different concentrations. Cytotoxicity was evaluated by MTT assay and Trypan blue dye exclusion assay. EEP at a concentration of 250 g/mL exhibited ≥50% mortality in all cell lines tested (i.e., IC50 value. EEP revealed a concentration and time dependent cytotoxic effect. Apoptosis was estimated by differential staining (ethidium bromide/acridine orange and TUNEL (deoxynucleotidyl transferase-dUTP nick end labeling assay. Light microscopy and atomic force microscopy demonstrated morphological features of apoptosis in all the cell lines after treatment with 250 g/mL EEP for 24 h. Thus, early onset of apoptosis is the reason for anticancer activity of Indian stingless bee propolis. Further, the antioxidant potential of Indian stingless bee propolis was demonstrated to substantiate its anticancer activity.

  7. High clusterin expression correlates with a poor outcome in stage II colorectal cancers.

    LENUS (Irish Health Repository)

    Kevans, David

    2012-02-01

    The role of clusterin in tumor growth and progression remains unclear. Overexpression of cytoplasmic clusterin has been studied in aggressive colon tumors; however, no correlation between clusterin expression and survival in colorectal cancer has been identified to date. We assessed levels of clusterin expression in a group of stage II colorectal cancer patients to assess its utility as a prognostic marker. The study included 251 patients with stage II colorectal cancer. Tissue microarrays were constructed and immunohistochemistry done and correlated with clinical features and long term outcome. Dual immunofluorescence and confocal microscopy were used with terminal deoxynucleotidyl-transferase-mediated dUTP nick-end labeling probes and clusterin antibody to assess the degree of co localization. Percentage epithelial cytoplasmic staining was higher in tumor compared with nonadjacent normal mucosa (P < 0.001). Within the stromal compartment, percentage cytoplamic staining and intensity was lower in tumor tissue compared with normal nonadjacent mucosa (P < or = 0.001). Survival was significantly associated with percentage epithelial cytoplasmic staining (P < 0.001), epithelial cytoplasmic staining intensity (P < 0.001), percentage stromal cytoplasmic staining (P = 0.002), and stromal cytoplasmic staining intensity (P < 0.001). Clusterin levels are associated with poor survival in stage II colorectal cancer.

  8. The Extract of Aster Koraiensis Prevents Retinal Pericyte Apoptosis in Diabetic Rats and Its Active Compound, Chlorogenic Acid Inhibits AGE Formation and AGE/RAGE Interaction

    Directory of Open Access Journals (Sweden)

    Junghyun Kim

    2016-09-01

    Full Text Available Retinal capillary cell loss is a hallmark of early diabetic retinal changes. Advanced glycation end products (AGEs are believed to contribute to retinal microvascular cell loss in diabetic retinopathy. In this study, the protective effects of Aster koraiensis extract (AKE against damage to retinal vascular cells were investigated in streptozotocin (STZ-induced diabetic rats. To examine this issue further, AGE accumulation, nuclear factor-kappaB (NF-κB and inducible nitric oxide synthase (iNOS were investigated using retinal trypsin digests from streptozotocin-induced diabetic rats. In the diabetic rats, TUNEL (Terminal deoxynucleotidyl transferase mediated dUTP Nick End Labeling-positive retinal microvascular cells were markedly increased. Immunohistochemical studies revealed that AGEs were accumulated within the retinal microvascular cells, and this accumulation paralleled the activation of NF-κB and the expression of iNOS in the diabetic rats. However, AKE prevented retinal microvascular cell apoptosis through the inhibition of AGE accumulation and NF-κB activation. Moreover, to determine the active compounds of AKE, two major compounds, chlorogenic acid and 3,5-di-O-caffeoylquinic acid, were tested in an in vitro assay. Among these compounds, chlorogenic acid significantly reduced AGE formation as well as AGE/RAGE (receptor for AGEs binding activity. These results suggest that AKE, particularly chlorogenic acid, is useful in inhibiting AGE accumulation in retinal vessels and exerts a preventive effect against the injuries of diabetic retinal vascular cells.

  9. Neuroprotective effect of nicorandil through inhibition of apoptosis by the PI3K/Akt1 pathway in a mouse model of deep hypothermic low flow.

    Science.gov (United States)

    Yu, Di; Fan, Changfeng; Zhang, Weiyan; Wen, Zhongyuan; Hu, Liang; Yang, Lei; Feng, Yu; Yin, Ke-Jie; Mo, Xuming

    2015-10-15

    Nicorandil exerts a protective effect on ischemia-reperfusion (I/R) injury in the brain and kidney through anti-apoptotic mechanisms. However, the mechanism by which nicorandil protects against I/R injury induced by deep hypothermic low flow (DHLF) remains unclear. We used a cerebral I/R model induced by DHLF to determine the neuroprotective effects and possible mechanisms of nicorandil. Hematoxylin-eosin (HE) staining and in situ terminal deoxynucleotidyl transferase UTP nick end labeling (TUNEL) assay were used to detect changes in cell morphology and the number of apoptotic cells in hippocampus, respectively. The apoptotic regulators including Bcl-2, Bax, Akt, and p-Akt (the active, phosphorylated form of Akt) were examined by Western blot (WB). Histopathological findings showed that nicorandil significantly alleviated morphological damage in hippocampal and reduced the number of TUNEL-positive nuclei induced by DHLF. Nicorandil also increased the expression of Bcl-2 and decreased the expression of Bax, while increasing p-Akt level. Consistent with these results, nicorandil-mediated neuroprotection was reduced in the Akt1+/- mutant mice and inhibited by LY294002, a PI3K inhibitor. These findings showed that nicorandil provides a neuroprotective role in DHLF-induced I/R injury by inhibiting apoptosis via activation of the PI3K/Akt1 signaling pathway. Copyright © 2015 Elsevier B.V. All rights reserved.

  10. An ethanolic extract of leaves of Piper betle (Paan) Linn mediates its antileishmanial activity via apoptosis.

    Science.gov (United States)

    Sarkar, Avijit; Sen, Rupashree; Saha, Piu; Ganguly, Sudipto; Mandal, Goutam; Chatterjee, Mitali

    2008-05-01

    An unprecedented increase in the incidence of unresponsiveness to antimonial compounds has highlighted the urgent need to develop new antileishmanial agents. The leaves of Piper betle (locally known as Paan) have long been in use in the Indian indigenous system of medicine for its antimicrobial properties but its antileishmanial potential has not been studied. Accordingly, an ethanolic extract of leaves of Piper betle (PB) was tested for its antileishmanial activity that was evidenced in both promastigotes and amastigotes, with IC50 values of 9.8 and 5.45 microg/ml, respectively; importantly, it was accompanied by a safety index of >12-fold. This leishmanicidal activity of PB was mediated via apoptosis as evidenced by morphological changes, loss of mitochondrial membrane potential, in situ labeling of DNA fragments by terminal deoxyribonucleotidyltransferase-mediated deoxyuridine triphosphate nick end labeling, and cell-cycle arrest at the sub-G0/G1 phase. Taken together, the data indicate that PB has promising antileishmanial activity that is mediated via programmed cell death and, accordingly, merits consideration and further investigation as a therapeutic option for the treatment of leishmaniasis.

  11. Aerosol-delivered programmed cell death 4 enhanced apoptosis, controlled cell cycle and suppressed AP-1 activity in the lungs of AP-1 luciferase reporter mice.

    Science.gov (United States)

    Hwang, S-K; Jin, H; Kwon, J T; Chang, S-H; Kim, T H; Cho, C-S; Lee, K H; Young, M R; Colburn, N H; Beck, G R; Yang, H-S; Cho, M-H

    2007-09-01

    The long-term survival of lung cancer patients treated with conventional therapies remains poor and therefore the need for novel approaches remains high. This has led to the re-emergence of aerosol delivery as a therapeutic intervention. In this study, glucosylated polyethylenimine (GPEI) was used as carrier to investigate programmed cell death 4 (PDCD4) and PDCD4 mutant (D418A), an eIF4A-binding mutant, on PDCD4-related signaling and activator protein-1 (AP-1) activity in the lungs of AP-1 luciferase reporter mice. After confirming the efficiency of GPEI as a carrier in lungs, the effects of aerosol-delivered PDCD4 were investigated in AP-1 luciferase reporter mice. Aerosol delivery of GPEI/PDCD4 through a nose-only inhalation facilitated the apoptosis of lungs whereas aerosol PDCD4 mutant did not. Also, such aerosol delivery regulated proteins relevant to cell-cycle control and suppressed AP-1 activity. Results obtained by western blot analysis, immunohistochemistry, luciferase assay and deoxynucleotidyl-transferase-mediated nick end labeling study suggest that combined actions such as facilitating apoptosis, controlling cell cycle and suppression of AP-1 activity by PDCD4 may provide useful tool for designing lung tumor prevention and treatment by which PDCD4 functions as a transformation suppressor in the future.

  12. DNA damage in preserved specimens and tissue samples: a molecular assessment

    Directory of Open Access Journals (Sweden)

    Cantin Elizabeth

    2008-10-01

    Full Text Available Abstract The extraction of genetic information from preserved tissue samples or museum specimens is a fundamental component of many fields of research, including the Barcode of Life initiative, forensic investigations, biological studies using scat sample analysis, and cancer research utilizing formaldehyde-fixed, paraffin-embedded tissue. Efforts to obtain genetic information from these sources are often hampered by an inability to amplify the desired DNA as a consequence of DNA damage. Previous studies have described techniques for improved DNA extraction from such samples or focused on the effect of damaging agents – such as light, oxygen or formaldehyde – on free nucleotides. We present ongoing work to characterize lesions in DNA samples extracted from preserved specimens. The extracted DNA is digested to single nucleosides with a combination of DNase I, Snake Venom Phosphodiesterase, and Antarctic Phosphatase and then analyzed by HPLC-ESI-TOF-MS. We present data for moth specimens that were preserved dried and pinned with no additional preservative and for frog tissue samples that were preserved in either ethanol, or formaldehyde, or fixed in formaldehyde and then preserved in ethanol. These preservation methods represent the most common methods of preserving animal specimens in museum collections. We observe changes in the nucleoside content of these samples over time, especially a loss of deoxyguanosine. We characterize the fragmentation state of the DNA and aim to identify abundant nucleoside lesions. Finally, simple models are introduced to describe the DNA fragmentation based on nicks and double-strand breaks.

  13. Production of DNA minicircles less than 250 base pairs through a novel concentrated DNA circularization assay enabling minicircle design with NF-κB inhibition activity

    Science.gov (United States)

    Thibault, Thomas; Degrouard, Jeril; Baril, Patrick; Pichon, Chantal; Midoux, Patrick

    2017-01-01

    Abstract Double-stranded DNA minicircles of less than 1000 bp in length have great interest in both fundamental research and therapeutic applications. Although minicircles have shown promising activity in gene therapy thanks to their good biostability and better intracellular trafficking, minicircles down to 250 bp in size have not yet been investigated from the test tube to the cell for lack of an efficient production method. Herein, we report a novel versatile plasmid-free method for the production of DNA minicircles comprising fewer than 250 bp. We designed a linear nicked DNA double-stranded oligonucleotide blunt-ended substrate for efficient minicircle production in a ligase-mediated and bending protein-assisted circularization reaction at high DNA concentration of 2 μM. This one pot multi-step reaction based-method yields hundreds of micrograms of minicircle with sequences of any base composition and position and containing or not a variety of site-specifically chemical modifications or physiological supercoiling. Biochemical and cellular studies were then conducted to design a 95 bp minicircle capable of binding in vitro two NF-κB transcription factors per minicircle and to efficiently inhibiting NF-κB-dependent transcriptional activity in human cells. Therefore, our production method could pave the way for the design of minicircles as new decoy nucleic acids. PMID:27899652

  14. Evaluation of human sperm chromatin status after selection using a modified Diff-Quik stain indicates embryo quality and pregnancy outcomes following in vitro fertilization.

    Science.gov (United States)

    Tavares, R S; Silva, A F; Lourenço, B; Almeida-Santos, T; Sousa, A P; Ramalho-Santos, J

    2013-11-01

    Sperm chromatin/DNA damage can be measured by a variety of assays. However, it has been reported that these tests may lose prognostic value in Assisted Reproductive Technology (ART) cycles when assessed in post-prepared samples, possibly due to the normalizing effect promoted by sperm preparation procedures. We have recently implemented a modified version of the Diff-Quik staining assay that allows for the evaluation of human sperm chromatin status in native samples, together with standard sperm morphology assessment. However, the value of this parameter in terms of predicting in vitro fertilization (IVF) and Intracytoplasmic sperm injection (ICSI) outcomes after sperm selection is unknown. In this study, data from 138 couples undergoing in vitro fertilization (IVF) or Intracytoplasmic sperm injection (ICSI) treatments showed that sperm chromatin integrity was significantly improved after density gradient centrifugation and swim up (p embryo development rates (p > 0.05). However, sperm samples presenting lower percentages of damaged chromatin were associated with better quality (Grade I) embryos in both ART procedures (p selection may occur; but not in ICSI, where sperm selection is operator dependent. This quick and low-cost assay is suggested as an alternative method to detect sperm chromatin status in minimal clinical settings, when no other well-established and robust assays (e.g. Sperm chromatin structure assay, terminal deoxynucleotidyl transferase-mediated dUDP nick-end labelling) are available. © 2013 American Society of Andrology and European Academy of Andrology.

  15. Overexpression of IL-7R alpha provides a competitive advantage during early T-cell development.

    Science.gov (United States)

    Laouar, Yasmina; Crispe, I Nicholas; Flavell, Richard A

    2004-03-15

    Critical checkpoints controlling early thymic T-cell development and homeostasis are set by the proper signaling function of the interleukin 7 receptor (IL-7R) and the pre-T-cell antigen receptor. Although alpha beta T-cell development is observed in IL-7- and IL-7R alpha-deficient mice, the number of thymocytes is significantly reduced, implying a role for the IL-7R in controlling the size of the thymic T-cell compartment. Here, we report the overexpression of IL-7R alpha that occurs in the early T-cell compartment from AKR/J mice, animals that are highly susceptible to the spontaneous development of thymoma. Increased IL-7R alpha was revealed by surface staining, and increased IL-7R alpha mRNA was documented by using reverse transcriptase-polymerase chain reaction (RT-PCR). This resulted in increased survival of AKR/J early thymocytes, shown by the decreased frequency of TUNEL(+) (terminal deoxynucleotidyl transferase mediated deoxyuridine triphosphate [dUTP]-fluorescein nick end labeling) cells. In an in vivo thymocyte repopulation model, AKR/J thymocytes had a selective advantage over healthy thymocytes. This advantage occurred at early stages of T-cell development. Our findings support the model that overexpression of growth factor receptors can contribute to proliferation and malignancy.

  16. Cell cycle arrest induced by inhibitors of epigenetic modifications in maize (Zea mays) seedling leaves: characterization of the process and possible mechanisms involved.

    Science.gov (United States)

    Wang, Pu; Zhang, Hao; Hou, Haoli; Wang, Qing; Li, Yingnan; Huang, Yan; Xie, Liangfu; Gao, Fei; He, Shibin; Li, Lijia

    2016-07-01

    Epigenetic modifications play crucial roles in the regulation of chromatin architecture and are involved in cell cycle progression, including mitosis and meiosis. To explore the relationship between epigenetic modifications and the cell cycle, we treated maize (Zea mays) seedlings with six different epigenetic modification-related inhibitors and identified the postsynthetic phase (G2 ) arrest via flow cytometry analysis. Total H4K5ac levels were significantly increased and the distribution of H3S10ph signalling was obviously changed in mitosis under various treatments. Further statistics of the cells in different periods of mitosis confirmed that the cell cycle was arrested at preprophase. Concentrations of hydrogen peroxide were relatively higher in the treated plants and the antioxidant thiourea could negate the influence of the inhibitors. Moreover, all of the treated plants displayed negative results in the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labelling (TUNEL) and γ-H2AX immunostaining assays after exposure for 3 d. Additionally, the expression level of topoisomerase genes in the treated plants was relatively lower than that in the untreated plants. These results suggest that these inhibitors of epigenetic modifications could cause preprophase arrest via reactive oxygen species formation inhibiting the expression of DNA topoisomerase genes, accompanied by changes in the H4K5ac and H3S10ph histone modifications. © 2016 The Authors. New Phytologist © 2016 New Phytologist Trust.

  17. Inhibitory effects of CP on the growth of human gastric adenocarcinoma BGC-823 tumours in nude mice.

    Science.gov (United States)

    Wang, Hai-Jun; Liu, Yu; Zhou, Bao-Jun; Zhang, Zhan-Xue; Li, Ai-Ying; An, Ran; Yue, Bin; Fan, Li-Qiao; Li, Yong

    2018-05-01

    Objective To investigate the potential antitumour effects of [2-(6-amino-purine-9-yl)-1-hydroxy-phosphine acyl ethyl] phosphonic acid (CP) against gastric adenocarcinoma. Methods Human BGC-823 xenotransplants were established in nude mice. Animals were randomly divided into control and CP groups, which were administered NaHCO 3 vehicle alone or CP dissolved in NaHCO 3 (200 µg/kg body weight) daily, respectively. Tumour volume was measured weekly for 6 weeks. Resected tumours were assayed for proliferative activity with anti-Ki-67 or anti-proliferating cell nuclear antigen (PCNA) antibodies. Cell apoptosis was examined using terminal deoxynucleotidyl transferase-mediated dUTP nick end labelling (TUNEL) assays and with caspase-3 immunostaining. Proteins were measured by Western blotting. Results There was a significant reduction in tumour volume and a reduced percentage of Ki-67-positive or PCNA-positive cells in the CP group compared with the control group. The percentage of TUNEL-positive or caspase 3-positive cells significantly increased following CP treatment compared with the control group. Tumours from the CP group had higher levels of phosphorylated-extracellular signal-regulated kinase (p-ERK) and phosphorylated-AKT (p-AKT) compared with control tumours. Conclusion CP treatment inhibited tumour growth and induced tumour cell apoptosis in a nude mouse model of BGC-823 gastric adenocarcinoma. Activation of the AKT and ERK signalling pathways may mediate this antitumour activity.

  18. Andrographis paniculata extract induced apoptosis of adenocarcinoma mammae in C3H mice

    Directory of Open Access Journals (Sweden)

    Nugrahaningsih

    2013-08-01

    Full Text Available BACKGROUND Apoptosis plays an important role in tumorigenesis. Induction of apoptosis is a strategy for developing cancer therapy. In vitro study found that andrographolide isolated from Andrographis paniculata has anticancer activity by an apoptotic mechanism in cancer cell lines. The aim of the present study was to prove the effect of Andrographis paniculata extract administered orally on apoptosis of mammary adenocarcinoma in C3H mice. METHODS This study was of post test randomized control group design. Twenty four C3H mice with transplanted mammary adenocarcinomas were divided into four groups. To three groups Andrographis paniculata extract was administered orally for 14 days, at doses of 5, 10 and 15 mg/day, respectively, whereas to the control group no Andrographis paniculata extract was administered. On day 15 the mice were terminated. The mammary adenocarcinomas were examined by the terminal deoxynucleotide transferase dUTP nick end labeling (TUNEL method. The values of the apoptotic index were expressed as mean±SD and analyzed using Anova and Pearson’s correlation test. RESULTS The mean apoptotic index values differed significantly among the experimental groups (p=0.001. The highest value was found in the group receiving Andrographis paniculata extract 15 mg/day, while the lowest was in the control group, the values being significantly correlated (r=0.974. CONCLUSIONS Oral administration of Andrographis paniculata extract induced apoptosis in C3H mice with mammary adenocarcinoma

  19. A ‘Paradox of Expression’: Merleau-Ponty and the Intertwining Nature of Brecht’s ‘not...but’ Procedure

    Directory of Open Access Journals (Sweden)

    Cohen Ambrose

    2017-06-01

    Full Text Available This article seeks to investigate the practical applications of a performance process that Bertolt Brecht called the procedure of “fixing the ‘not…but’,” which produces a Verfremdungseffekt. The article also interrogates the philosophy that such a process inherently performs. In one of his last writings, Maurice Merleau-Ponty argues that to look at oneself through the eyes of another necessarily blends the divide between one body and another and, by applying one’s senses to another’s, one engages in a “paradox of expression.” I explore the process of working with actors to produce distancing effects in their acting by chronicling the results of a 2012 practice-as-research project titled The Galileo Experiment. I use Merleau-Ponty’s “paradox of expression” as a way of considering Brecht’s call for the co-presence of the actor and their character in a stage performance. I borrow Nick Crossley’s approach to phenomenological intersubjectivity and consider other theoretical implications of performing the ‘not...but’ procedure. I argue that in order for the actor to successfully perform Brecht’s ‘not...but’ procedure, the actor must play into their character while occasionally playing out of the character in an alternative attitude, using what I call the ‘reflective block’.

  20. Lesion-induced DNA weak structural changes detected by pulsed EPR spectroscopy combined with site-directed spin labelling.

    Science.gov (United States)

    Sicoli, Giuseppe; Mathis, Gérald; Aci-Sèche, Samia; Saint-Pierre, Christine; Boulard, Yves; Gasparutto, Didier; Gambarelli, Serge

    2009-06-01

    Double electron-electron resonance (DEER) was applied to determine nanometre spin-spin distances on DNA duplexes that contain selected structural alterations. The present approach to evaluate the structural features of DNA damages is thus related to the interspin distance changes, as well as to the flexibility of the overall structure deduced from the distance distribution. A set of site-directed nitroxide-labelled double-stranded DNA fragments containing defined lesions, namely an 8-oxoguanine, an abasic site or abasic site analogues, a nick, a gap and a bulge structure were prepared and then analysed by the DEER spectroscopic technique. New insights into the application of 4-pulse DEER sequence are also provided, in particular with respect to the spin probes' positions and the rigidity of selected systems. The lesion-induced conformational changes observed, which were supported by molecular dynamics studies, confirm the results obtained by other, more conventional, spectroscopic techniques. Thus, the experimental approaches described herein provide an efficient method for probing lesion-induced structural changes of nucleic acids.

  1. Structure-guided mutational analysis of the nucleotidyltransferase domain of Escherichia coli NAD+-dependent DNA ligase (LigA).

    Science.gov (United States)

    Zhu, Hui; Shuman, Stewart

    2005-04-01

    NAD+-dependent DNA ligase (LigA) is essential for bacterial growth and a potential target for antimicrobial drug discovery. Here we queried the role of 14 conserved amino acids of Escherichia coli LigA by alanine scanning and thereby identified five new residues within the nucleotidyltransferase domain as being essential for LigA function in vitro and in vivo. Structure activity relationships were determined by conservative mutagenesis for the Glu-173, Arg-200, Arg-208, and Arg-277 side chains, as well as four other essential side chains that had been identified previously (Lys-115, Asp-117, Asp-285, and Lys-314). In addition, we identified Lys-290 as important for LigA activity. Reference to the structure of Enterococcus faecalis LigA allowed us to discriminate three classes of essential/important side chains that: (i) contact NAD+ directly (Lys-115, Glu-173, Lys-290, and Lys-314); (ii) comprise the interface between the NMN-binding domain (domain Ia) and the nucleotidyltransferase domain or comprise part of a nick-binding site on the surface of the nucleotidyltransferase domain (Arg-200 and Arg-208); or (iii) stabilize the active site fold of the nucleotidyltransferase domain (Arg-277). Analysis of mutational effects on the isolated ligase adenylylation and phosphodiester formation reactions revealed different functions for essential side chains at different steps of the DNA ligase pathway, consistent with the proposal that the active site is serially remodeled as the reaction proceeds.

  2. Inhibitors of nuclease and redox activity of apurinic/apyrimidinic endonuclease 1/redox effector factor 1 (APE1/Ref-1).

    Science.gov (United States)

    Laev, Sergey S; Salakhutdinov, Nariman F; Lavrik, Olga I

    2017-05-01

    Human apurinic/apyrimidinic endonuclease 1/redox effector factor 1 (APE1/Ref-1) is a multifunctional protein which is essential in the base excision repair (BER) pathway of DNA lesions caused by oxidation and alkylation. This protein hydrolyzes DNA adjacent to the 5'-end of an apurinic/apyrimidinic (AP) site to produce a nick with a 3'-hydroxyl group and a 5'-deoxyribose phosphate moiety or activates the DNA-binding activity of certain transcription factors through its redox function. Studies have indicated a role for APE1/Ref-1 in the pathogenesis of cancer and in resistance to DNA-interactive drugs. Thus, this protein has potential as a target in cancer treatment. As a result, major efforts have been directed to identify small molecule inhibitors against APE1/Ref-1 activities. These agents have the potential to become anticancer drugs. The aim of this review is to present recent progress in studies of all published small molecule APE1/Ref-1 inhibitors. The structures and activities of APE1/Ref-1 inhibitors, that target both DNA repair and redox activities, are presented and discussed. To date, there is an urgent need for further development of the design and synthesis of APE1/Ref-1 inhibitors due to high importance of this protein target. Copyright © 2017 Elsevier Ltd. All rights reserved.

  3. A Cytotoxic Hydroperoxy Sterol from the Brown Alga, Nizamuddinia Zanardinii

    Directory of Open Access Journals (Sweden)

    Abdolhossein Rustaiyan

    2013-03-01

    Full Text Available Background:The marine environment is a unique source of bioactive natural products, of which Nizamuddinia zanardinii is an important brown algae distributed in Oman Sea. Literature revealed that there is no report on phytochemistry and pharmacology of this valuable algae.Methods:Bioguided fractionation of the methanolic extract of Nizamuddinia zanardinii, collected from Oman Sea, led to the isolation of a hydroperoxy sterol. Its structure was determined by analysis of the spectroscopic data as 24-hydroperoxy-24-vinyl cholesterol (HVC. In vitro cytotoxic activity of this compound was evaluated against HT29, MCF7, A549, HepG2 and MDBK cell lines.Results:Although 24(R-hydroproxy-24-vinylcholesterol has been previously reported from Sargassum and Padina species, it is the first report on the presence of this compound from N. zanardinii. This compound exhibited cytotoxicity in all cell lines (IC50, 3.62, 9.09, 17.96, 32.31 and 37.31 μg/mL respectively. HVC was also evaluated for apoptotic activity and demonstrated positive results in terminal deoxynucleotidyl transferase dUTP Nick End labeling (TUNEL assay suggesting it a candidate for further apoptotic studies.Conclusions:Nizamuddinia zanardinii, a remarkable brown algae of Oman Sea, is a good source of hydroproxy sterols with promising cytotoxic on various cell lines particularly human colon adenocarcinoma.

  4. Histone Modification Is Involved in Okadaic Acid (OA Induced DNA Damage Response and G2-M Transition Arrest in Maize.

    Directory of Open Access Journals (Sweden)

    Hao Zhang

    Full Text Available Histone modifications are involved in regulation of chromatin structure. To investigate the relationship between chromatin modification and cell cycle regulation during plant cell proliferation, Okadaic acid (OA, a specific inhibitor of serine/threonine protein phosphatase, was applied in this study. The results showed that OA caused the cell cycle arrest at preprophase, leading to seedling growth inhibition. Western blotting assay revealed that the spatial distribution of phosphorylation of Ser10 histone H3 tails (H3S10ph signals was altered under OA treatment. Reactive oxygen species (ROS was found to be at higher levels and TdT-mediated dUTP nick end labeling (TUNEL assay displayed DNA breaks happened at the chromatin after treatment with OA, companied with an increase in the acetylation of histone H4 at lysine 5 (H4K5ac level. From these observations, we speculated that the alteration of the spatial distribution of H3S10ph and the level of H4K5ac was involved in the procedure that OA induced DNA breaks and G2-M arrested by the accumulation of ROS, and that the histone H3S10ph and H4K5ac might facilitate DNA repair by their association with the chromatin decondensation.

  5. Ciné Club

    CERN Multimedia

    Ciné Club

    2010-01-01

    Thursday 23 September 2010 at 20:30 CERN Main Auditorium Lock, Stock and Two Smoking Barrels By: Guy Ritchie (UK, 1998) 107 min With/avec: Jason Flemyng, Dexter Fletcher, Nick Moran, Jason Statham   Four London working class stiffs pool their money to put one in a high stakes card game, but things go wrong and they end up owing half a million pounds and having one week to come up with the cash. Overhearing their neighbours in the next flat plotting to hold up a group of out-of-their-depth drug growers, our heros decide to stitch up the robbers in turn. In a way the confusion really starts when a pair of antique double-barrelled shotguns go missing in a completely different scam.   Original version english with english subtitles Entrance : 2 CHF Projection from DVD http://cineclub.web.cern.ch/Cineclub/   Thursday 30 September 2010 at 20:30 CERN Main Auditorium Facing Windows / La finestra di fronte By/de : Ferzan Ozpetek (Italy, 2003) 106 min With/avec: Giovanna Mezz...

  6. Production of DNA minicircles less than 250 base pairs through a novel concentrated DNA circularization assay enabling minicircle design with NF-κB inhibition activity.

    Science.gov (United States)

    Thibault, Thomas; Degrouard, Jeril; Baril, Patrick; Pichon, Chantal; Midoux, Patrick; Malinge, Jean-Marc

    2017-03-17

    Double-stranded DNA minicircles of less than 1000 bp in length have great interest in both fundamental research and therapeutic applications. Although minicircles have shown promising activity in gene therapy thanks to their good biostability and better intracellular trafficking, minicircles down to 250 bp in size have not yet been investigated from the test tube to the cell for lack of an efficient production method. Herein, we report a novel versatile plasmid-free method for the production of DNA minicircles comprising fewer than 250 bp. We designed a linear nicked DNA double-stranded oligonucleotide blunt-ended substrate for efficient minicircle production in a ligase-mediated and bending protein-assisted circularization reaction at high DNA concentration of 2 μM. This one pot multi-step reaction based-method yields hundreds of micrograms of minicircle with sequences of any base composition and position and containing or not a variety of site-specifically chemical modifications or physiological supercoiling. Biochemical and cellular studies were then conducted to design a 95 bp minicircle capable of binding in vitro two NF-κB transcription factors per minicircle and to efficiently inhibiting NF-κB-dependent transcriptional activity in human cells. Therefore, our production method could pave the way for the design of minicircles as new decoy nucleic acids. © The Author(s) 2016. Published by Oxford University Press on behalf of Nucleic Acids Research.

  7. Carbon market emerges

    International Nuclear Information System (INIS)

    Hordern, N.

    2001-01-01

    Last November COP6, the UN climate change conference in The Hague, failed to agree on the rules of the Kyoto Protocol, the treaty limiting developed countries' emissions of greenhouse gases ('carbon'). Many in the Australian resource and energy sector were relieved that COP6 was inconclusive. As Industry Minister Senator Nick Minchin put it: 'Better no outcome than a bad outcome.' However, the financial services sector -potentially a major beneficiary of the international carbon it hoped COP6 would endorse -received a setback. Apparently not for long. Only months later, a nascent Australian carbon trading market seems to be emerging by stealth, irrespective of COP6's fortunes. This poses both opportunities and costs for the resources and energy sectors. Whoever succeeds in influencing the design of a trading scheme, 'pilot' or not, will have a box seat when - not if - a 'mandatory' emissions trading scheme is introduced. Government policy is that there will be no mandatory emissions trading scheme until the establishment of a UN endorsed international market requires one of Australia. This, in turn, depends upon the conclusion of the negotiations that stalled in The Hague. COP6 is set to resume in Germany, probably in July

  8. Integrated Observations From Fixed and AUV Platforms in the Littoral Zone at the SFOMC Coastal Ocean Observatory

    Science.gov (United States)

    Dhanak, M. R.

    2001-12-01

    A 12-hour survey of the coastal waters off the east coast of Florida at the South Florida Ocean Measurement Center (SFOMC) coastal ocean observatory, during summer 1999, is described to illustrate the observatory's capabilities for ocean observation. The facility is located close to the Gulf Stream, the continental shelf break being only 3 miles from shore and is therefore influenced by the Gulf Stream meanders and the instability of the horizontal shear layer at its edge. As a result, both cross-shelf and along-shelf components of currents in the littoral zone can undergo dramatic +/- 0.5 m/s oscillations. Observations of surface currents from an OSCR, and of subsurface structure from an autonomous underwater vehicle (AUV) platform, a bottom-mounted ADCP and CT-chain arrays during the survey will be described and compared. The AUV on-board sensors included upward and downward looking 1200kHz ADCP, a CTD package and a small-scale turbulence package, consisting of two shear probes and a fast-response thermistor. Prevailing atmospheric conditions were recorded at an on-site buoy. The combined observations depict flows over a range of scales. Acknowledgements: The observations from the OSCR are due to Nick Shay and Tom Cook (University of Miami), and from the bottom-mounted ADCP, CT chain arrays and the surface buoy are due to Alex Soloviev (Nova Southeastern University) and Mark Luther and Bob Weisberg (University of South Florida).

  9. Ultrastructural characterization of the meiotic prophase. A tool in the assessment of radiation damage in man

    Energy Technology Data Exchange (ETDEWEB)

    Holm, P.B.; Rasmussen, S.W.; von Wettstein, D. (Carlsberg Lab., Copenhagen (Denmark). Dept. of Physiology)

    1982-01-01

    The three-dimensional reconstruction of meiotic nuclei from serial sections micrographed in the electron microscope has provided information about man and several other organisms that is not obtainable by light microscopy or biochemical analysis. At zygotene, the previously unpaired chromosomes align and form synaptonemal complexes between homologous chromosome segments either by progressive initiation from the telomeres or by interstitial recognition. Chromosome and bivalent interlocking at zygotene is a regular phenomenon and occurs at a frequency of 0.7-4.0 per nucleus in samples of meiocytes analyzed from different organisms. This frequency is reduced to 0.1 per nucleus at pachytene. The interlockings are resolved by breakage and precise rejoining of the broken ends. This breakage and rejoining can also occur in the absence of the DNA nicking and repair involved in crossing-over. The synaptonemal complexes combining homologous chromosome segments are stabilized by recombination nodules, after which a second round of synaptonemal complex formation between as yet unpaired or unstably paired chromosome segments occurs, apparently for optimization of bivalent formation. Nonhomologous pairing with the synaptonemal complex can take place in this phase of pachytene.

  10. Effect of SMAD7 gene overexpression on TGF-β1-induced profibrotic responses in fibroblasts derived from Peyronie's plaque

    Directory of Open Access Journals (Sweden)

    Min Ji Choi

    2015-06-01

    Full Text Available Transforming growth factor-β1 (TGF-β1 has been identified as one of the most important fibrogenic cytokines associated with Peyronie's disease (PD. The mothers against decapentaplegic homolog 7 (SMAD7 is an inhibitory Smad protein that blocks TGF-β signaling pathway. The aim of this study was to examine the anti-fibrotic effect of the SMAD7 gene in primary fibroblasts derived from human PD plaques. PD fibroblasts were pretreated with the SMAD7 gene and then stimulated with TGF-β1. Treated fibroblasts were used for Western blotting, fluorescent immunocytochemistry, hydroxyproline determination, and terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling assays. Overexpression of the SMAD7 gene inhibited TGF-β1-induced phosphorylation and nuclear translocation of SMAD2 and SMAD3, transdifferentiation of fibroblasts into myofibroblasts, and quashed TGF-β1-induced production of extracellular matrix protein and hydroxyproline. Overexpression of the SMAD7 gene decreased the expression of cyclin D1 (a positive cell cycle regulator and induced the expression of poly (ADP-ribose polymerase 1, which is known to terminate Smad-mediated transcription, in PD fibroblasts. These findings suggest that the blocking of the TGF-β pathway by use of SMAD7 may be a promising therapeutic strategy for the treatment of PD.

  11. Active Site Sharing and Subterminal Hairpin Recognition in a New Class of DNA Transposases

    Energy Technology Data Exchange (ETDEWEB)

    Ronning, Donald R.; Guynet, Catherine; Ton-Hoang, Bao; Perez, Zhanita N.; Ghirlando, Rodolfo; Chandler, Michael; Dyda, Fred (Centre Nat); (NIH)

    2010-07-20

    Many bacteria harbor simple transposable elements termed insertion sequences (IS). In Helicobacter pylori, the chimeric IS605 family elements are particularly interesting due to their proximity to genes encoding gastric epithelial invasion factors. Protein sequences of IS605 transposases do not bear the hallmarks of other well-characterized transposases. We have solved the crystal structure of full-length transposase (TnpA) of a representative member, ISHp608. Structurally, TnpA does not resemble any characterized transposase; rather, it is related to rolling circle replication (RCR) proteins. Consistent with RCR, Mg{sup 2+} and a conserved tyrosine, Tyr127, are essential for DNA nicking and the formation of a covalent intermediate between TnpA and DNA. TnpA is dimeric, contains two shared active sites, and binds two DNA stem loops representing the conserved inverted repeats near each end of ISHp608. The cocrystal structure with stem-loop DNA illustrates how this family of transposases specifically recognizes and pairs ends, necessary steps during transposition.

  12. A mitochondria-dependent pathway mediates the apoptosis of GSE-induced yeast.

    Directory of Open Access Journals (Sweden)

    Sishuo Cao

    Full Text Available Grapefruit seed extract (GSE, which has powerful anti-fungal activity, can induce apoptosis in S. cerevisiae. The yeast cells underwent apoptosis as determined by testing for apoptotic markers of DNA cleavage and typical chromatin condensation by Terminal Deoxynucleotidyl Transferase-mediated dUTP Nick End Labeling (TUNEL and 4,6'-diaminidino-2-phenylindole (DAPI staining and electron microscopy. The changes of ΔΨmt (mitochondrial transmembrane potential and ROS (reactive oxygen species indicated that the mitochondria took part in the apoptotic process. Changes in this process detected by metabonomics and proteomics revealed that the yeast cells tenaciously resisted adversity. Proteins related to redox, cellular structure, membrane, energy and DNA repair were significantly increased. In this study, the relative changes in the levels of proteins and metabolites showed the tenacious resistance of yeast cells. However, GSE induced apoptosis in the yeast cells by destruction of the mitochondrial 60 S ribosomal protein, L14-A, and prevented the conversion of pantothenic acid to coenzyme A (CoA. The relationship between the proteins and metabolites was analyzed by orthogonal projections to latent structures (OPLS. We found that the changes of the metabolites and the protein changes had relevant consistency.

  13. Enhanced healing of mitomycin C-treated healing-impaired wounds in rats with PRP-containing fragmin/protamine microparticles (PRP&F/P MPs).

    Science.gov (United States)

    Takikawa, Megumi; Ishihara, Masayuki; Takabayashi, Yuki; Sumi, Yuki; Takikawa, Makoto; Yoshida, Ryuichi; Nakamura, Shingo; Hattori, Hidemi; Yanagibayashi, Satoshi; Yamamoto, Naoto; Kiyosawa, Tomoharu

    2015-04-13

    The purpose of this study was to evaluate the accelerating effects of platelet-rich plasma-containing (PRP&) fragmin/protamine microparticles (F/P MPs) for repairing mitomycin C-treated healing-impaired wounds. Staining with terminal deoxynucleotidyl transferase-mediated dUTP nick-end labelling (TUNEL-staining) showed that apoptosis of dermal fibroblast cells (DFCs) and epidermal keratinocyte cells (EKCs) were significantly induced in the skin of the mitomycin C-treated rats. Full-thickness skin defects were made on the back of rats and mitomycin C was applied on the wounds to prepare a healing-impaired wound. After washing out the mitomycin C, saline (control), F/P MPs alone, PRP alone, and PRP&F/P MPs were injected around the wounds. The rats were later euthanised and histological sections of the wounds were then prepared at indicated time periods after the treatment. These results indicated the numbers of large, medium, and small capillary lumens 7 days after injection of PRP&F/P MPs were significantly higher than those after injection of PRP or F/P MPs alone. Furthermore, epithelium and granulation tissue formations were significantly stimulated in the healing-impaired wounds treated with PRP&F/P MPs 3, 7 and 14 days after injection of PRP&F/P MPs.

  14. Effect of green light spectra on the reduction of retinal damage and stress in goldfish, Carassius auratus

    International Nuclear Information System (INIS)

    Song, Jin Ah; Kim, Na Na; Choi, Young Jae; Choi, Cheol Young

    2016-01-01

    We investigated the effect of light spectra on retinal damage and stress in goldfish using green (530 nm) and red (620 nm) light emitting diodes (LEDs) at three intensities each (0.5, 1.0, and 1.5 W/m"2). We measured the change in the levels of plasma cortisol and H_2O_2 and expression and levels of caspase-3. The apoptotic response of green and red LED spectra was assessed using the terminal transferase dUTP nick end labeling (TUNEL) assay. Stress indicator (cortisol and H_2O_2) and apoptosis-related genes (caspase-3) decreased in green light, but increased in red light with higher light intensities over time. The TUNEL assay revealed that more apoptotic cells were detected in outer nuclear layers after exposure to red LED over time with the increase in light intensity, than the other spectra. These results indicate that green light efficiently reduces retinal damage and stress, whereas red light induces it. Therefore, red light-induced retina damage may induce apoptosis in goldfish retina. -- Highlights: •Green light efficiently reduces retinal damage and stress. •Green spectra reduce caspase production and apoptosis. •Red light-induced retina damage may induce apoptosis in goldfish retina. •The retina of goldfish recognizes green spectra as a stable environment.

  15. Fluorine-induced apoptosis and lipid peroxidation in human hair follicles in vitro.

    Science.gov (United States)

    Wang, Zheng-hui; Li, Xiao-li; Yang, Zhuang-qun; Xu, Min

    2010-12-01

    Fluoride is an essential trace element for human body; however, exposure to high amounts of fluoride has been documented to be correlated with an increasing risk of hair loss. To date, little is known about the mechanism(s) of how fluoride affects hair follicles. Here, we demonstrated that middle (1.0 mmol/L) and high (10.0 mmol/L) concentrations of sodium fluoride (NaF) significantly inhibited hair follicle elongation in vitro, but low NaF (0.1 mmol/L) showed little influence. Moreover, treatment with high levels of NaF resulted in a marked increase in terminal dUTP nick end labeling-positive cells in the outer layer of the outer root sheath, the dermal sheath, and the lower bulb matrix surrounding dermal papilla. Furthermore, the enhanced apoptosis was coupled with an increased oxidative stress manifested as higher malondialdehyde content. Additionally, the presence of selenium considerably antagonized the effects of middle NaF on hair follicles, with regard to either the suppression of hair growth or the induction of oxidative stress and apoptosis. In conclusion, exposure to high levels of fluoride compromises hair follicle growth and accelerate cell apoptosis in vitro. The toxicity of fluoride can be reduced by selenium, at least partially via the suppression of intracellular oxidative stress.

  16. Stimulatory effects of curcumin and quercetin on posttranslational modifications of p53 during lung carcinogenesis.

    Science.gov (United States)

    Zhang, P; Zhang, Xy

    2018-06-01

    Lung cancer is responsible for increase in mortality due to cancer-related deaths, and new approaches are being explored for the betterment of the situation. In the present study, chemopreventive efficacy of curcumin and quercetin was investigated against benzo(a)pyrene (BP)-induced lung carcinogenesis. The mice were segregated into five groups, which included normal control, BP-treated, BP + curcumin-treated, BP + quercetin-treated, and BP + curcumin + quercetin-treated groups. The morphological and histological analyses of tumor nodules confirmed lung carcinogenesis22 weeks after weeks single intraperitoneal injection of BP at a dose of 100 mg/kg body weight to mice. Curcumin and quercetin when administered individually as well as in combination significantly elevated the expression of acetylated-p53, which was otherwise depressed due to BP treatment. Also, both the phytochemicals significantly reduced the BP-inflicted increased levels of phosphorylated-p53. Furthermore, observed increase in the number of apoptotic cells by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL), assay and increased activities of caspase 3 and 9 confirmed the induction of apoptosis by curcumin and quercetin. Moreover, the histological slides also showed noticeable improvement in the histoarchitecture of lungs by phytochemicals. The present study concludes that prophylactic treatment with curcumin and quercetin induces apoptosis in the lungs by modulation of p53 posttranslational modifications.

  17. Extracellular Matrix and Growth Factors Improve the Efficacy of Intramuscular Islet Transplantation.

    Directory of Open Access Journals (Sweden)

    Haruyuki Tsuchiya

    Full Text Available The efficacy of intramuscular islet transplantation is poor despite being technically simple, safe, and associated with reduced rates of severe complications. We evaluated the efficacy of combined treatment with extracellular matrix (ECM and growth factors in intramuscular islet transplantation.Male BALB/C mice were used for the in vitro and transplantation studies. The following three groups were evaluated: islets without treatment (islets-only group, islets embedded in ECM with growth factors (Matrigel group, and islets embedded in ECM without growth factors [growth factor-reduced (GFR Matrigel group]. The viability and insulin-releasing function of islets cultured for 96 h were significantly improved in Matrigel and GFR Matrigel groups compared with the islets-only group.Blood glucose and serum insulin levels immediately following transplantation were significantly improved in the Matrigel and GFR Matrigel groups and remained significantly improved in the Matrigel group at postoperative day (POD 28. On histological examination, significantly decreased numbers of TdT-mediated deoxyuridine triphosphate-biotin nick end labeling-positive islet cells and significantly increased numbers of Ki67-positive cells were observed in the Matrigel and GFR Matrigel groups at POD 3. Peri-islet revascularization was most prominent in the Matrigel group at POD 14.The efficacy of intramuscular islet transplantation was improved by combination treatment with ECM and growth factors through the inhibition of apoptosis, increased proliferation of islet cells, and promotion of revascularization.

  18. Mapping vaccinia virus DNA replication origins at nucleotide level by deep sequencing.

    Science.gov (United States)

    Senkevich, Tatiana G; Bruno, Daniel; Martens, Craig; Porcella, Stephen F; Wolf, Yuri I; Moss, Bernard

    2015-09-01

    Poxviruses reproduce in the host cytoplasm and encode most or all of the enzymes and factors needed for expression and synthesis of their double-stranded DNA genomes. Nevertheless, the mode of poxvirus DNA replication and the nature and location of the replication origins remain unknown. A current but unsubstantiated model posits only leading strand synthesis starting at a nick near one covalently closed end of the genome and continuing around the other end to generate a concatemer that is subsequently resolved into unit genomes. The existence of specific origins has been questioned because any plasmid can replicate in cells infected by vaccinia virus (VACV), the prototype poxvirus. We applied directional deep sequencing of short single-stranded DNA fragments enriched for RNA-primed nascent strands isolated from the cytoplasm of VACV-infected cells to pinpoint replication origins. The origins were identified as the switching points of the fragment directions, which correspond to the transition from continuous to discontinuous DNA synthesis. Origins containing a prominent initiation point mapped to a sequence within the hairpin loop at one end of the VACV genome and to the same sequence within the concatemeric junction of replication intermediates. These findings support a model for poxvirus genome replication that involves leading and lagging strand synthesis and is consistent with the requirements for primase and ligase activities as well as earlier electron microscopic and biochemical studies implicating a replication origin at the end of the VACV genome.

  19. Arsenic induced apoptosis in rat liver following repeated 60 days exposure

    International Nuclear Information System (INIS)

    Bashir, Somia; Sharma, Yukti; Irshad, M.; Nag, T.C.; Tiwari, Monica; Kabra, M.; Dogra, T.D.

    2006-01-01

    Background: Accumulation of the wide spread environmental toxin arsenic in liver results in hepatotoxcity. Exposure to arsenite and other arsenicals has been previously shown to induce apoptosis in certain tumor cell lines at low (1-3 μM) concentration. Aim: The present study was focused to elucidate the role of free radicals in arsenic toxicity and to investigate the nature of in vivo sodium arsenite induced cell death in liver. Methods: Male wistar rats were exposed to arsenite at three different doses of 0.05, 2.5 and 5 mg/l for 60 days. Oxidative stress in liver was measured by estimating pro-oxidant and antioxidant activity in liver. Histopathological examination of liver was carried out by light and transmission electron microscopy. Analysis of DNA fragmentation by gel electrophoresis was used to identify apoptosis after the exposure. Terminal deoxy-nucleotidyl transferase mediated dUTP Nick end-labeling (TUNEL) assay was used to qualify and quantify apoptosis. Results: A significant increase in cytochrome-P450 and lipid peroxidation accompanied with a significant alteration in the activity of many of the antioxidants was observed, all suggestive of arsenic induced oxidative stress. Histopathological examination under light and transmission electron microscope suggested a combination of ongoing necrosis and apoptosis. DNA-TUNEL showed an increase in apoptotic cells in liver. Agarose gel electrophoresis of DNA of hepatocytes resulted in a characteristic ladder pattern. Conclusion: Chronic arsenic administration induces a specific pattern of apoptosis called post-mitotic apoptosis

  20. Curcumin attenuates palmitate-induced apoptosis in MIN6 pancreatic β-cells through PI3K/Akt/FoxO1 and mitochondrial survival pathways.

    Science.gov (United States)

    Hao, Feng; Kang, Jinsen; Cao, Yajun; Fan, Shengjun; Yang, Haopeng; An, Yu; Pan, Yan; Tie, Lu; Li, Xuejun

    2015-11-01

    Lipotoxicity plays a vital role in development and progression of type 2 diabetes. Prolonged elevation of free fatty acids especially the palmitate leads to pancreatic β-cell dysfunction and apoptosis. Curcumin (diferuloylmethane), a polyphenol from the curry spice turmeric, is considered to be a broadly cytoprotective agent. The present study was designed to determine the protective effect of curcumin on palmitate-induced apoptosis in β-cells and investigate underlying mechanisms. Our results showed that curcumin improved cell viability and enhanced glucose-induced insulin secretory function in MIN6 pancreatic β-cells. Palmitate incubation evoked chromatin condensation, DNA nick end labeling and activation of caspase-3 and -9. Curcumin treatment inhibited palmitate-induced apoptosis, relieved mitochondrial depolarization and up-regulated Bcl-2/Bax ratio. Palmitate induced the generation of reactive oxygen species and inhibited activities of antioxidant enzymes, which could be neutralized by curcumin treatment. Moreover, curcumin could promote rapid phosphorylation of Akt and nuclear exclusion of FoxO1 in MIN6 cells under lipotoxic condition. Phosphatidylinositol 3-kinase and Akt specific inhibitors abolished the anti-lipotoxic effect of curcumin and stimulated FoxO1 nuclear translocation. These findings suggested that curcumin protected MIN6 pancreatic β-Cells against apoptosis through activation of Akt, inhibition of nuclear translocation of FoxO1 and mitochondrial survival pathway.

  1. Detection of radiation-induced apoptosis using the comet assay

    International Nuclear Information System (INIS)

    Wada, Seiichi; Kobayashi, Yasuhiko; Funayama, Tomoo; Yamamoto, Kazuo; Khoa, Tran Van; Natsuhori, Masahiro; Ito, Nobuhiko

    2003-01-01

    The electrophoresis pattern of apoptotic cells detected by the comet assay has a characteristic small head and spread tail. This image has been referred to as an apoptotic comet, but it has not been previously proven to be apoptotic cells by any direct method. In order to identify this image obtained by the comet assay as corresponding to an apoptotic cell, the frequency of appearance of apoptosis was examined using CHO-K1 and L5178Y cells which were exposed to gamma irradiation. As a method for detecting apoptosis, the terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) assay was used. When the frequency of appearance of apoptotic cells following gamma irradiation was observed over a period of time, there was a significant increase in appearance of apoptosis when using the TUNEL assay. However, there was only a slight increase when using the comet assay. In order to verify the low frequency of appearance of apoptosis when using the comet assay, we attempted to use the TUNEL assay to satin the apoptotic comets detected in the comet assay. The apoptotic comets were TUNEL positive and the normal comets were TUNEL negative. This indicates that the apoptotic comets were formed from DNA fragments with 3'-hydroxy ends that are generated as cells undergo apoptosis. Therefore, it was understood that the characteristic pattern of apoptotic comets detected by the comet assay corresponds to cells undergoing apoptosis. (author)

  2. Texto, textualidades e jornalismo: a legitimação de práticas culturais nos editoriais do Diário Folha de Rondônia

    Directory of Open Access Journals (Sweden)

    Colferai, Sandro Adalberto

    2010-01-01

    Full Text Available Este artigo aborda a tomada de sentidos preferenciais em textos jornalísticos produzidos e em circulação na região amazônica. O recorte é feito a partir de textos de editoriais publicados pelo jornal diário Folha de Rondônia, editado no interior de Rondônia. A análise dos textos é feita a partir da perspectiva dos Estudos Culturais, como apresentada por Stuart Hall, Richard Johnson e Nick Couldry. São tomados os conceitos de Cultura, Identidade Cultural e Texto, além de se recuperar parte da história social da Amazônia e, em especial, de Rondônia, esta imersa no choque entre diferentes práticas simbólicas de populações tradicionais da Amazônia e colonizadores que passaram a ocupá-la a partir da década de 1970

  3. Ischemia preconditioning is neuroprotective in a rat cerebral ischemic injury model through autophagy activation and apoptosis inhibition

    International Nuclear Information System (INIS)

    Xia, D.Y.; Li, W.; Qian, H.R.; Yao, S.; Liu, J.G.; Qi, X.K.

    2013-01-01

    Sublethal ischemic preconditioning (IPC) is a powerful inducer of ischemic brain tolerance. However, its underlying mechanisms are still not well understood. In this study, we chose four different IPC paradigms, namely 5 min (5 min duration), 5×5 min (5 min duration, 2 episodes, 15-min interval), 5×5×5 min (5 min duration, 3 episodes, 15-min intervals), and 15 min (15 min duration), and demonstrated that three episodes of 5 min IPC activated autophagy to the greatest extent 24 h after IPC, as evidenced by Beclin expression and LC3-I/II conversion. Autophagic activation was mediated by the tuberous sclerosis type 1 (TSC1)-mTor signal pathway as IPC increased TSC1 but decreased mTor phosphorylation. Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) and hematoxylin and eosin staining confirmed that IPC protected against cerebral ischemic/reperfusion (I/R) injury. Critically, 3-methyladenine, an inhibitor of autophagy, abolished the neuroprotection of IPC and, by contrast, rapamycin, an autophagy inducer, potentiated it. Cleaved caspase-3 expression, neurological scores, and infarct volume in different groups further confirmed the protection of IPC against I/R injury. Taken together, our data indicate that autophagy activation might underlie the protection of IPC against ischemic injury by inhibiting apoptosis

  4. Ablation of phosphoinositide 3-kinase-gamma reduces the severity of acute pancreatitis.

    Science.gov (United States)

    Lupia, Enrico; Goffi, Alberto; De Giuli, Paolo; Azzolino, Ornella; Bosco, Ornella; Patrucco, Enrico; Vivaldo, Maria Cristina; Ricca, Marco; Wymann, Matthias P; Hirsch, Emilio; Montrucchio, Giuseppe; Emanuelli, Giorgio

    2004-12-01

    In pancreatic acini, the G-protein-activated phosphoinositide 3-kinase-gamma (PI3K gamma) regulates several key pathological responses to cholecystokinin hyperstimulation in vitro. Thus, using mice lacking PI3K gamma, we studied the function of this enzyme in vivo in two different models of acute pancreatitis. The disease was induced by supramaximal concentrations of cerulein and by feeding mice a choline-deficient/ethionine-supplemented diet. Although the secretive function of isolated pancreatic acini was identical in mutant and control samples, in both models, genetic ablation of PI3K gamma significantly reduced the extent of acinar cell injury/necrosis. In agreement with a protective role of apoptosis in pancreatitis, PI3K gamma-deficient pancreata showed an increased number of apoptotic acinar cells, as determined by terminal dUTP nick-end labeling and caspase-3 activity. In addition, neutrophil infiltration within the pancreatic tissue was also reduced, suggesting a dual action of PI3K gamma, both in the triggering events within acinar cells and in the subsequent neutrophil recruitment and activation. Finally, the lethality of the choline-deficient/ethionine-supplemented diet-induced pancreatitis was significantly reduced in mice lacking PI3K gamma. Our results thus suggest that inhibition of PI3K gamma may be of therapeutic value in acute pancreatitis.

  5. Ablation of Phosphoinositide 3-Kinase-γ Reduces the Severity of Acute Pancreatitis

    Science.gov (United States)

    Lupia, Enrico; Goffi, Alberto; De Giuli, Paolo; Azzolino, Ornella; Bosco, Ornella; Patrucco, Enrico; Vivaldo, Maria Cristina; Ricca, Marco; Wymann, Matthias P.; Hirsch, Emilio; Montrucchio, Giuseppe; Emanuelli, Giorgio

    2004-01-01

    In pancreatic acini, the G-protein-activated phosphoinositide 3-kinase-γ (PI3Kγ) regulates several key pathological responses to cholecystokinin hyperstimulation in vitro. Thus, using mice lacking PI3Kγ, we studied the function of this enzyme in vivo in two different models of acute pancreatitis. The disease was induced by supramaximal concentrations of cerulein and by feeding mice a choline-deficient/ethionine-supplemented diet. Although the secretive function of isolated pancreatic acini was identical in mutant and control samples, in both models, genetic ablation of PI3Kγ significantly reduced the extent of acinar cell injury/necrosis. In agreement with a protective role of apoptosis in pancreatitis, PI3Kγ-deficient pancreata showed an increased number of apoptotic acinar cells, as determined by terminal dUTP nick-end labeling and caspase-3 activity. In addition, neutrophil infiltration within the pancreatic tissue was also reduced, suggesting a dual action of PI3Kγ, both in the triggering events within acinar cells and in the subsequent neutrophil recruitment and activation. Finally, the lethality of the choline-deficient/ethionine-supplemented diet-induced pancreatitis was significantly reduced in mice lacking PI3Kγ. Our results thus suggest that inhibition of PI3Kγ may be of therapeutic value in acute pancreatitis. PMID:15579443

  6. Determination of trace amounts of nickel(II) with α-(2-Benzimidazolyl)-α',α''-(N-5-nitro-2-pyridyl hydrazone) toluene in the presence of triton X-100 by fluorescence method

    International Nuclear Information System (INIS)

    Park, Chan Il; Kim, Hyun Soo; Cha, Ki Won

    2000-01-01

    A method is described for the fluorimetric determination of nicke, based on the formation of Ni(II)-α-(2-Benzimidazolyl)-α',α''-(N-5-nitro-2-pyridyl hydrazone)-toluene complex in the presence of a non-ionic surfactant. The complex has practically no fluorescence in the absence of surfactant, but the addition of Triton X-100 makes possible the fluorimetric determination of low concentrations of Ni(II) as it enhances the fluorescence intensity of the complex by up to about 5-fold. This method is very sensitive and selective for the direct determination of nickel ion. The optimum conditions are a Triton X-100 concentration of 2.0 mL (5.0%, v/v) and pH 9.0 ± 0.2 (ammonium chloride-ammonia buffer). The fluorescence is measured at 337 nm of emission wavelength under 300 nm of excitation wavelength. The fluorescence intensity is a linear function of the concentration of Ni(II) in the range 5-70 ng/mL, and the detection limit is 2.0 ng/mL. The proposed method has been successfully applied to the determination of trace amounts of Ni(II) in food and human hair samples

  7. Vedolizumab: toward a personalized therapy paradigm for people with ulcerative colitis

    Directory of Open Access Journals (Sweden)

    Dart RJ

    2017-03-01

    Full Text Available Robin J Dart,1–3,* Mark A Samaan,1,* Nick Powell,1,4 Peter M Irving1 1Department of Gastroenterology, IBD Centre, St. Thomas’ Hospital, 2Immunosurveillance Laboratory, The Francis Crick Institute, London, 3Division of Immunology, Infection, and Inflammatory Disease (DIIID, King’s College London, 4Department of Experimental Immunobiology, Division of Transplantation and Mucosal Biology, King’s College London, London, UK *These authors contributed equally to this work Abstract: Ulcerative colitis (UC is a chronic relapsing and remitting inflammatory bowel disease, with a characteristic leukocytic infiltration of the mucosa. Immunosuppression including anti-TNF-α therapy is a mainstay of treatment for many; however, systemic immunosuppression is not universally effective and is associated with potential side effects. The gut-tropic integrin α4β7, which is expressed on leukocytes, mediates migration from the circulation to the intestinal mucosa. Vedolizumab is a monoclonal antibody which blocks the egress of leukocytes via α4β7, preventing accumulation in the mucosa, and attenuating inflammation without systemic immunosuppression. Vedolizumab has been evaluated in UC in a phase III trial, demonstrating efficacy as both an induction and a maintenance agent. In this article, we review the clinical trial data and also explore the growing body of “real-world” effectiveness data, investigating response and remission rates of vedolizumab in clinical practice. In addition, we review the increasing volume of data supporting the reassuring safety profile associated with vedolizumab. Keywords: vedolizumab, ulcerative colitis, personalized therapy

  8. Edaravone Protect against Retinal Damage in Streptozotocin-Induced Diabetic Mice

    Science.gov (United States)

    Liu, Xiaoyi; Chen, Xi; Xie, Ping; Yuan, Songtao; Zhang, Weiwei; Lin, Xiaojun; Liu, Qinghuai

    2014-01-01

    Edaravone (3-methyl-1-phenyl-2-pyrazolin-5-one), a free radical scavenger, is used for the clinical treatment of retinal injury. In this study, we investigated the protective effects of edaravone against diabetic retinal damage in the mouse. Diabetic retinopathy in the mouse was induced by injection of streptozotocin. Edaravone was given once-daily and was intraperitoneally (i.p.) treated at a dose of 3 mg/kg from streptozotocin injection to 4 weeks after onset of diabetes. Retinal ganglion cells (RGCs) damage was evaluated by recording the pattern electroretinogram (ERG). RGCs damage was also detected by Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining, and the levels of reactive oxygen species (ROS) were determined fluorometrically. The expressions of phosporylated-ERK1/2, BDNF, and caspase-3 were determined by Western blot analysis. Retinal levels of ROS, phosphorylated ERK1/2, and cleaved caspase-3 were significantly increased, whereas the expression of BDNF was significantly decreased in the retinas of diabetic mice, compared to nondiabetic mice. Administration of edaravone significantly attenuated diabetes induced RGCs death, upregulation of ROS, ERK1/2 phosphorylation, and cleaved caspase-3 and downregulation of BDNF. These findings suggest that oxidative stress plays a pivotal role in diabetic retinal damage and that systemic administration of edaravone may slow the progression of retinal neuropathy induced by diabetes. PMID:24897298

  9. Edaravone protect against retinal damage in streptozotocin-induced diabetic mice.

    Directory of Open Access Journals (Sweden)

    Dongqing Yuan

    Full Text Available Edaravone (3-methyl-1-phenyl-2-pyrazolin-5-one, a free radical scavenger, is used for the clinical treatment of retinal injury. In this study, we investigated the protective effects of edaravone against diabetic retinal damage in the mouse. Diabetic retinopathy in the mouse was induced by injection of streptozotocin. Edaravone was given once-daily and was intraperitoneally (i.p. treated at a dose of 3 mg/kg from streptozotocin injection to 4 weeks after onset of diabetes. Retinal ganglion cells (RGCs damage was evaluated by recording the pattern electroretinogram (ERG. RGCs damage was also detected by Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL staining, and the levels of reactive oxygen species (ROS were determined fluorometrically. The expressions of phosporylated-ERK1/2, BDNF, and caspase-3 were determined by Western blot analysis. Retinal levels of ROS, phosphorylated ERK1/2, and cleaved caspase-3 were significantly increased, whereas the expression of BDNF was significantly decreased in the retinas of diabetic mice, compared to nondiabetic mice. Administration of edaravone significantly attenuated diabetes induced RGCs death, upregulation of ROS, ERK1/2 phosphorylation, and cleaved caspase-3 and downregulation of BDNF. These findings suggest that oxidative stress plays a pivotal role in diabetic retinal damage and that systemic administration of edaravone may slow the progression of retinal neuropathy induced by diabetes.

  10. Proceedings of the 2003 NASA/JPL Workshop on Fundamental Physics in Space

    Science.gov (United States)

    Strayer, Don (Editor)

    2003-01-01

    The 2003 Fundamental Physics workshop included presentations ranging from forces acting on RNA to properties of clouds of degenerate Fermi atoms, to techniques to probe for a added space-time dimensions, and to flight hardware for low temperature experiments, amongst others. Mark Lee from NASA Headquarters described the new strategic plan that NASA has developed under Administrator Sean O'Keefe's leadership. Mark explained that the Fundamental Physics community now needs to align its research program and the roadmap describing the long-term goals of the program with the NASA plan. Ulf Israelsson of JPL discussed how the rewrite of the roadmap will be implemented under the leadership of the Fundamental Physics Discipline Working Group (DWG). Nick Bigelow, chair of the DWG, outlined how investigators can contribute to the writing of the roadmap. Results of measurements on very cold clouds of Fermi atoms near a Feshbach resonance were described by three investigators. Also, new measurements relating to tests of Einstein equivalence were discussed. Investigators also described methods to test other aspects of Einstein's relativity theories.

  11. Crystal structure of a Fanconi anemia-associated nuclease homolog bound to 5' flap DNA: basis of interstrand cross-link repair by FAN1

    Energy Technology Data Exchange (ETDEWEB)

    Gwon, Gwang Hyeon; Kim, Youngran; Liu, Yaqi; Watson, Adam T.; Jo, Aera; Etheridge, Thomas J.; Yuan, Fenghua; Zhang, Yanbin; Kim, YoungChang; Carr, Anthony M.; Cho, Yunje

    2014-10-15

    Fanconi anemia (FA) is an autosomal recessive genetic disorder caused by defects in any of 15 FA genes responsible for processing DNA interstrand cross-links (ICLs). The ultimate outcome of the FA pathway is resolution of cross-links, which requires structure-selective nucleases. FA-associated nuclease 1 (FAN1) is believed to be recruited to lesions by a monoubiquitinated FANCI–FANCD2 (ID) complex and participates in ICL repair. Here, we determined the crystal structure of Pseudomonas aeruginosa FAN1 (PaFAN1) lacking the UBZ (ubiquitin-binding zinc) domain in complex with 5' flap DNA. All four domains of the right-hand-shaped PaFAN1 are involved in DNA recognition, with each domain playing a specific role in bending DNA at the nick. The six-helix bundle that binds the junction connects to the catalytic viral replication and repair (VRR) nuclease (VRR nuc) domain, enabling FAN1 to incise the scissile phosphate a few bases distant from the junction. The six-helix bundle also inhibits the cleavage of intact Holliday junctions. PaFAN1 shares several conserved features with other flap structure-selective nucleases despite structural differences. A clamping motion of the domains around the wedge helix, which acts as a pivot, facilitates nucleolytic cleavage. The PaFAN1 structure provides insights into how archaeal Holliday junction resolvases evolved to incise 5' flap substrates and how FAN1 integrates with the FA complex to participate in ICL repair.

  12. [Influence of dendritic cell infiltration on prognosis and biologic characteristics of progressing gastric cancer].

    Science.gov (United States)

    Huang, Hai-li; Wu, Ben-yan; You, Wei-di; Shen, Ming-shi; Wang, Wen-ju

    2003-09-01

    To study the relation between dendritic cell (DC) infiltration and clinicopathologic parameters, biologic characteristics and prognosis of progressing gastric cancer. The development of apoptotic cell death (apoptotic index, AI) in 61 progressing gastric carcinoma tissues was analyzed by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate biotin nick end labeling (TUNEL) method. The PCNA labeling index (PCNA-LI), density of dendritic cells in the tumor were detected by immunohistochemical method by the LSAB kit using antibody against S-100 protein and PC-10. DC infiltration was negatively correlated with lymph node metastasis, clinical stage and PCNA-LI, but positively with AI. The DCs in gastric cancer groups with and without lymph node metastasis were (5.63 +/- 4.37)/HPF and (8.51 +/- 5.57)/HPF with difference significant (P stage lesions were (11.23 +/- 6.05)/HPF, (6.28 +/- 4.37)/HPF and (5.53 +/- 5.19)/HPF also with differences significant (P gastric carcinoma.

  13. Exposure to cigarette smoke increases apoptosis in the rat gastric mucosa through a reactive oxygen species-mediated and p53-independent pathway.

    Science.gov (United States)

    Wang, H; Ma, L; Li, Y; Cho, C H

    2000-04-01

    Cigarette smoking is a major risk factor for gastric cancer and peptic ulcer. The aim of our study was to investigate the relationship between exposure to cigarette smoke and apoptosis in the rat gastric mucosa and the mechanism involved. Rats were exposed to different concentrations of cigarette smoke (0, 2, and 4%) once daily for a different number of 1 h periods (1, 3, 6, and 9 d). Apoptosis was identified by the terminal deoxy-transferase (TdT)-mediated dUTP-biotin nick end labeling (TUNEL) method and caspase-3 activity. The mucosal xanthine oxidase (XO) activity and p53 level were also measured. The results showed that exposure to cigarette smoke produced a time- and concentration-dependent increase in apoptosis in the rat gastric mucosa that was accompanied by an increase in XO activity. The increased apoptosis and XO activity could be detected after even a single exposure. In contrast, the level of p53 was elevated only in the later stage of cigarette smoke exposure. The apoptotic effect could be blocked by pretreatment with an XO inhibitor (allopurinol, 20 mg/kg intraperitoneally) or a hydroxyl free radical scavenger (DMSO, 0.2%, 1 ml/kg intravenously). However, neither of these treatments had any effect on the p53 level of the mucosa. In summary, we conclude that exposure to cigarette smoke can increase apoptosis in the rat gastric mucosa through a reactive oxygen species- (ROS) mediated and a p53-independent pathway.

  14. Apoptosis and cell proliferation in the development of gastric carcinomas: associations with c-myc and p53 protein expression.

    Science.gov (United States)

    Ishii, Hideaki H; Gobé, Glenda C; Pan, Wenshen; Yoneyama, Juichi; Ebihara, Yoshiro

    2002-09-01

    Patients with gastric carcinomas have a poor prognosis and low survival rates. The aim of the present paper was to characterize cellular and molecular properties to provide insight into aspects of tumor progression in early compared with advanced gastric cancers. One hundred and nine graded gastric carcinomas (early or advanced stage, undifferentiated or differentiated type) with paired non-cancer tissue were studied to define the correlation between apoptosis (morphology, terminal deoxyribonucleotidyl transferase-mediated dUTP-digoxigenin nick end-labeling), cell proliferation (Ki-67 expression, morphology) and expression and localization of two proteins frequently having altered expression in cancers, namely p53 and c-myc. Overall, apoptosis was lower in early stage, differentiated and undifferentiated gastric carcinomas compared with advanced-stage cancers. Cell proliferation was comparatively high in all stages. There was a high level of p53 positivity in all stages. Only the early- and advanced-stage undifferentiated cancers that were p53 positive had a significantly higher level of apoptosis (P cancers that had either c-myc or p53-positivity. The results indicate that low apoptosis and high cell proliferation combine to drive gastric cancer development. The molecular controls for high cell proliferation of the early stage undifferentiated gastric cancers involve overexpression of both p53 and c-myc. Overexpression of p53 may also control cancer development in that its expression is associated with higher levels of apoptosis in early and late-stage undifferentiated, cancers. Copyright 2002 Blackwell Publishing Asia Pty Ltd

  15. Expression of caspase-3 predicts prognosis in advanced noncardia gastric cancer.

    Science.gov (United States)

    Amptoulach, Sousana; Lazaris, Andreas C; Giannopoulou, Ioanna; Kavantzas, Nikolaos; Patsouris, Efstratios; Tsavaris, Nikolaos

    2015-01-01

    There is strong evidence that tumor growth is not only a result of uncontrolled cell proliferation but also of decreased apoptosis. Caspase-3 is a member of interleukin-1 beta-converting enzyme which is involved in the induction of apoptosis. Data on the expression of caspase-3 in patients with gastric cancer and its association with patient outcome are somewhat contradictory. We aimed to investigate the potential relation of the expression of caspase-3 protein with response to therapy and overall survival in patients with advanced noncardia gastric cancer. Tumor tissue samples collected from 359 consecutive patients with gastric cancer stage IV were retrospectively analyzed for the expression of caspase-3 in the primary tumor. The DNA apoptotic index assessed by the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-biotin nick end labeling method. All patients were followed up until death. Caspase-3 was expressed in 43.5 % of tumors. Caspase-3 expression compared to no expression was related with a higher DNA apoptotic index (p gastric cancer is a predictor of poor response to treatment and survival. Further studies are needed to fully elucidate the prognostic value of caspase-3 expression in these patients.

  16. The relationship between apoptosis and the expression of proliferating cell nuclear antigen and the clinical stages in gastric carcinoma.

    Science.gov (United States)

    Tao, K; Chen, D; Tian, Y; Lu, X; Yang, X

    2000-01-01

    The relationship between the apoptosis and the expression of proliferating cell nuclear antigen (PCNA) and the clinical stages in gastric cancers was studied. By using terminal deoxynucleotidyl transferase-mediated nick end labeling (TUNEL) technique and PCNA immunohistochemical staining, the apoptosis and the expression of PCNA in tissue of gastric carcinoma were assayed in situ, the index of apoptosis (AI), index of PCNA (PI) and the rate of AI/PI were calculated. AI and PI in gastric cancer tissues were (6.5 +/- 3.7)% and (49.8 +/- 15.9)% respectively, and the rate of AI/PI was 0.13 +/- 0.05, which were obviously different from those of normal gastric mucosa in paragastric cancer (P stages of gastric carcinoma, the AI was decreased, PI was increased and the rate of AI/PI decreased in gastric carcinoma. There was significant difference in them between the gastric cancer tissues and normal gastric mucosa in pericarcinoma in TNM stage II to IV (P gastric carcinoma. The AI, PI and the rate of AI/PI would become the prognostic factors in advanced gastric carcinoma.

  17. Prognostic significance of Fas and Fas ligand system-associated apoptosis in gastric cancer.

    Science.gov (United States)

    Ohno, S; Tachibana, M; Shibakita, M; Dhar, D K; Yoshimura, H; Kinugasa, S; Kubota, H; Masunaga, R; Nagasue, N

    2000-12-01

    Previous studies indicate that gastric carcinomas express Fas ligand and down-regulate Fas to escape from the host immune attack; however, the prognostic importance of Fas/FasL expression in this tumor is yet to be evaluated. Specimens from 87 gastric carcinoma patients of different stages treated in a defined period with curative intent were evaluated for apoptosis, Fas, FasL, and CD8 expression using an immunohistochemical method. The percentage of terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL)-positive apoptotic cells expressed as apoptotic index (AI) was higher in 43 patients when the cut-off value was set at the median value. There were no significant correlations between AI and clinicopathologic parameters. Thirty-nine patients showed a high number of CD8+ cells within cancer nests. Positive FasL and Fas expression was seen in 53 and 72 patients, respectively. CD8 and FasL expressions were related only to patients' age. Fas expression had significant correlations with tumor invasion and Lauren classification. There were significant direct correlations between AI and number of nest CD8+ cells and between AI and grade of Fas expression. Apoptotic index, pT stage, CD8 expression, and Fas expression were identified as independent prognostic factors. Spontaneous apoptosis in gastric carcinoma may be an independent prognosticator for survival and is significantly influenced by tumor Fas expression and number of nest CD8 + cells.

  18. Evidence that expression of a mutated p53 gene attenuates apoptotic cell death in human gastric intestinal-type carcinomas in vivo.

    Science.gov (United States)

    Ishida, M; Gomyo, Y; Ohfuji, S; Ikeda, M; Kawasaki, H; Ito, H

    1997-05-01

    To examine in vivo the validity of the results of experiments in vitro, we analyzed the relationship between p53 gene status and apoptotic cell death of human gastric intestinal-type adenocarcinomas. Surgical specimens were classified into two categories: 18 gastric cancers with nuclear p53 protein (A), and 17 gastric cancers without nuclear p53 protein (B). Polymerase chain reaction-single strand conformation polymorphism disclosed a shifted band that corresponded to a mutation in the p53 gene in 13 cases (72%) in category A and 3 cases (18%) in category B, the frequency being significantly higher in the former (P terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL). The TUNEL index [TI; (the number of TUNEL-positive apoptotic cells/the total number of tumor cells) x 100] was 3.8 +/- 1.4% in category A and 4.9 +/- 1.2% in category B, the value being significantly lower in the former (P gastric cancer, in accordance with the previous in vitro finding that p53 gene mutation provides a possible selective advantage for tumor cell proliferation, and (2) apoptosis is related not only to expression of p53 and the stage of the cell cycle, but also to p53-independent and cell cycle-independent events.

  19. Ethanol extract of Portulaca Oleracea L. reduced the carbon tetrachloride induced liver injury in mice involving enhancement of NF-κB activity

    Science.gov (United States)

    Shi, Hongguang; Liu, Xuefeng; Tang, Gusheng; Liu, Haiyan; Zhang, Yinghui; Zhang, Bo; Zhao, Xuezhi; Wang, Wanyin

    2014-01-01

    Acute hepatic injury causes high morbidity and mortality world-wide. Management of severe acute hepatic failure continues to be one of the most challenging problems in clinical medicine. In present study, carbon tetrachloride (CCl4) was used to induce acute liver damage in mice and the protective effects of ethanol extract of Portulaca Oleracea L. (PO) were examined. The aminotransferase activities were biochemical estimated and the liver damage was tested by morphological histological analysis and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay. The role of PO on the activity of NF-κB was determined by luciferase reporter gene assay and immunohistochemistry. The level of p-p65 was tested by western blot. Our results showed that PO administration on mice would decrease the serum aminotransferase level and reduced the liver histological damage. We also found that nuclear translocation of p65 was enhanced in liver tissues of mice treated with PO compared with control animals. In addition, in cultured hepatic cells, PO increased the NF-κB luciferase reporter gene activity and upregulated the level of phosphorylation of p65, but had no effects on mice liver SOD activity and MDA level. Collectively, PO attenuated CCl4 induced mice liver damage by enhancement of NF-κB activity. PMID:25628785

  20. Thimerosal-Derived Ethylmercury Is a Mitochondrial Toxin in Human Astrocytes: Possible Role of Fenton Chemistry in the Oxidation and Breakage of mtDNA

    Directory of Open Access Journals (Sweden)

    Martyn A. Sharpe

    2012-01-01

    Full Text Available Thimerosal generates ethylmercury in aqueous solution and is widely used as preservative. We have investigated the toxicology of Thimerosal in normal human astrocytes, paying particular attention to mitochondrial function and the generation of specific oxidants. We find that ethylmercury not only inhibits mitochondrial respiration leading to a drop in the steady state membrane potential, but also concurrent with these phenomena increases the formation of superoxide, hydrogen peroxide, and Fenton/Haber-Weiss generated hydroxyl radical. These oxidants increase the levels of cellular aldehyde/ketones. Additionally, we find a five-fold increase in the levels of oxidant damaged mitochondrial DNA bases and increases in the levels of mtDNA nicks and blunt-ended breaks. Highly damaged mitochondria are characterized by having very low membrane potentials, increased superoxide/hydrogen peroxide production, and extensively damaged mtDNA and proteins. These mitochondria appear to have undergone a permeability transition, an observation supported by the five-fold increase in Caspase-3 activity observed after Thimerosal treatment.

  1. Interstrand cross-links arising from strand breaks at true abasic sites in duplex DNA.

    Science.gov (United States)

    Yang, Zhiyu; Price, Nathan E; Johnson, Kevin M; Wang, Yinsheng; Gates, Kent S

    2017-06-20

    Interstrand cross-links are exceptionally bioactive DNA lesions. Endogenous generation of interstrand cross-links in genomic DNA may contribute to aging, neurodegeneration, and cancer. Abasic (Ap) sites are common lesions in genomic DNA that readily undergo spontaneous and amine-catalyzed strand cleavage reactions that generate a 2,3-didehydro-2,3-dideoxyribose sugar remnant (3'ddR5p) at the 3'-terminus of the strand break. Interestingly, this strand scission process leaves an electrophilic α,β-unsaturated aldehyde residue embedded within the resulting nicked duplex. Here we present evidence that 3'ddR5p derivatives generated by spermine-catalyzed strand cleavage at Ap sites in duplex DNA can react with adenine residues on the opposing strand to generate a complex lesion consisting of an interstrand cross-link adjacent to a strand break. The cross-link blocks DNA replication by ϕ29 DNA polymerase, a highly processive polymerase enzyme that couples synthesis with strand displacement. This suggests that 3'ddR5p-derived cross-links have the potential to block critical cellular DNA transactions that require strand separation. LC-MS/MS methods developed herein provide powerful tools for studying the occurrence and properties of these cross-links in biochemical and biological systems. © The Author(s) 2017. Published by Oxford University Press on behalf of Nucleic Acids Research.

  2. Possible Causes of Ileal Injury in Two Models of Microbial Sepsis and Protective Effect of Phytic Acid

    Directory of Open Access Journals (Sweden)

    Rasha Rashad Ahmed

    2010-03-01

    Full Text Available Background: Sepsis related-multiple organ dysfunction is associatedwith ileum injury. We aimed to determine the causes ofileal injury in two models of microbial sepsis resulted from infectionwith Aeromonas hydrophila or its endotoxin. We alsoevaluated the protective effect of phytic acid.Methods: Thin sections of ileum from 60 Swiss male mice incontrol, bacteria-infected or lipopolysaccharides (LPS andbacteria-infected or LPS-infected co-administered with phyticacid were subjected to histopathological and TdT-mediateddUTP nick-end labeling (TUNEL assay for apoptotic cellsdetection while ultra thin sections were stained with uranylacetate and lead citrate for cytological changes examination.Also, ileum images were exposed to the image analysis softwareto determine some related morphometric measures.Results: Necrosis and apoptosis were observed in ileum injuryin both examined sepsis models. The ileum injury was moresevere in LPS model. Phytic acid showed the ability to attenuateileum injury in Aeromonas hydrophila and its endotoxinmodels of sepsis after four weeks administration where itssupplementation significantly minimized the histopathologicaland cytological complications and morphometric alterationsresulted from the injury.Conclusion: The protective effects of phytic acid may becaused by increased mucous secretion, decreased apoptoticindex, attenuating the inflammatory and lymphocytic cellscount or increasing the renewal of the crypt cells and villousepithelial cells proliferation.

  3. Rapid and inexpensive method for isolating plasmid DNA

    International Nuclear Information System (INIS)

    Aljanabi, S. M.; Al-Awadi, S. J.; Al-Kazaz, A. A.; Baghdad Univ.

    1997-01-01

    A small-scale and economical method for isolating plasmid DNA from bacteria is described. The method provides DNA of suitable quality for most DNA manipulation techniques. This DNA can be used for restriction endonuclease digestion, southern blot hybridization, nick translation and end labeling of DNA probes, Polymerase Chain Reaction (PCR) -based techniques, transformation, DNA cycle-sequencing, and Chain-termination method for DNA sequencing. The entire procedure is adapted to 1.5 ml microfuge tubes and takes approximately 30 mins. The DNA isolated by this method has the same purity produced by CTAB and cesium chloride precipitation and purification procedures respectively. The two previous methods require many hours to obtain the final product and require the use of very expensive equipment as ultracentrifuge. This method is well suited for the isolation of plasmid DNA from a large number of bacterial samples and in a very short time and low cost in laboratories where chemicals, expensive equipment and finance are limited factors in conducting molecular research. (authors). 11refs. 11refs

  4. Role of oxidative stress in methamphetamine-induced dopaminergic toxicity mediated by protein kinase Cδ.

    Science.gov (United States)

    Shin, Eun-Joo; Duong, Chu Xuan; Nguyen, Xuan-Khanh Thi; Li, Zhengyi; Bing, Guoying; Bach, Jae-Hyung; Park, Dae Hun; Nakayama, Keiichi; Ali, Syed F; Kanthasamy, Anumantha G; Cadet, Jean Lud; Nabeshima, Toshitaka; Kim, Hyoung-Chun

    2012-06-15

    This study examined the role of protein kinase C (PKC) isozymes in methamphetamine (MA)-induced dopaminergic toxicity. Multiple-dose administration of MA did not significantly alter PKCα, PKCβI, PKCβII, or PKCζ expression in the striatum, but did significantly increase PKCδ expression. Gö6976 (a co-inhibitor of PKCα and -β), hispidin (PKCβ inhibitor), and PKCζ pseudosubstrate inhibitor (PKCζ inhibitor) did not significantly alter MA-induced behavioral impairments. However, rottlerin (PKCδ inhibitor) significantly attenuated behavioral impairments in a dose-dependent manner. In addition, MA-induced behavioral impairments were not apparent in PKCδ knockout (-/-) mice. MA-induced oxidative stress (i.e., lipid peroxidation and protein oxidation) was significantly attenuated in rottlerin-treated mice and was not apparent in PKCδ (-/-) mice. Consistent with this, MA-induced apoptosis (i.e., terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling-positive apoptotic cells) was significantly attenuated in rottlerin-treated mice. Furthermore, MA-induced increases in the dopamine (DA) turnover rate and decreases in tyrosine hydroxylase (TH) activity and the expression of TH, dopamine transporter (DAT), and vesicular monoamine transporter 2 (VMAT2) were not significantly observed in rottlerin-treated or PKCδ (-/-) mice. Our results suggest that PKCδ gene expression is a key mediator of oxidative stress and dopaminergic damage induced by MA. Thus, inhibition of PKCδ may be a useful target for protection against MA-induced neurotoxicity. Copyright © 2012 Elsevier B.V. All rights reserved.

  5. Current research on methamphetamine-induced neurotoxicity: animal models of monoamine disruption.

    Science.gov (United States)

    Kita, Taizo; Wagner, George C; Nakashima, Toshikatsu

    2003-07-01

    Methamphetamine (METH)-induced neurotoxicity is characterized by a long-lasting depletion of striatal dopamine (DA) and serotonin as well as damage to striatal dopaminergic and serotonergic nerve terminals. Several hypotheses regarding the mechanism underlying METH-induced neurotoxicity have been proposed. In particular, it is thought that endogenous DA in the striatum may play an important role in mediating METH-induced neuronal damage. This hypothesis is based on the observation of free radical formation and oxidative stress produced by auto-oxidation of DA consequent to its displacement from synaptic vesicles to cytoplasm. In addition, METH-induced neurotoxicity may be linked to the glutamate and nitric oxide systems within the striatum. Moreover, using knockout mice lacking the DA transporter, the vesicular monoamine transporter 2, c-fos, or nitric oxide synthetase, it was determined that these factors may be connected in some way to METH-induced neurotoxicity. Finally a role for apoptosis in METH-induced neurotoxicity has also been established including evidence of protection of bcl-2, expression of p53 protein, and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL), activity of caspase-3. The neuronal damage induced by METH may reflect neurological disorders such as autism and Parkinson's disease.

  6. Melatonin attenuates oxidative stress, liver damage and hepatocyte apoptosis after bile-duct ligation in rats.

    Science.gov (United States)

    Aktas, Cevat; Kanter, Mehmet; Erboga, Mustafa; Mete, Rafet; Oran, Mustafa

    2014-10-01

    The goal of this study was to evaluate the possible protective effects of melatonin against cholestatic oxidative stress, liver damage and hepatocyte apoptosis in the common rats with bile duct ligation (BDL). A total of 24 male Wistar albino rats were divided into three groups: control, BDL and BDL + received melatonin; each group contains eight animals. Melatonin-treated BDL rats received daily melatonin 100 mg/kg/day via intraperitoneal injection. The application of BDL clearly increased the malondialdehyde (MDA) levels and decreased the superoxide dismutase (SOD) and glutathione (GSH) activities. Melatonin treatment significantly decreased the elevated tissue MDA levels and increased the reduced SOD and GSH enzyme levels in the tissues. The changes demonstrate that the bile duct proliferation and fibrosis in expanded portal tracts include the extension of proliferated bile ducts into lobules, mononuclear cells and neutrophil infiltration into the widened portal areas as observed in the BDL group. The data indicate that melatonin attenuates BDL-induced cholestatic liver injury, bile duct proliferation and fibrosis. The α-smooth muscle actin (α-SMA) and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL)-positive cells in the BDL were observed to be reduced with the melatonin treatment. These results suggest that administration of melatonin is a potentially beneficial agent to reduce liver damage in BDL by decreasing oxidative stress. © The Author(s) 2012.

  7. DNA complexes with Ni nanoparticles: structural and functional properties

    Energy Technology Data Exchange (ETDEWEB)

    Tatarinova, Olga N.; Smirnov, Igor P. [Research Institute for Physico-Chemical Medicine of the Federal Medical-Biological Agency of the Russian Federation (Russian Federation); Safenkova, Irina V. [A.N. Bach Institute of Biochemistry (Russian Federation); Varizhuk, Anna M.; Pozmogova, Galina E., E-mail: pozmge@gmail.com [Research Institute for Physico-Chemical Medicine of the Federal Medical-Biological Agency of the Russian Federation (Russian Federation)

    2012-10-15

    Supramolecular complexes of biopolymers based on magnetic nanoparticles play an important role in creation of biosensors, implementation of theragnostic and gene therapeutic methods and biosafety evaluation. We investigated the impact of DNA interactions with nanoparticles of nickel (nNi) on the integrity and functionality of DNA. Data obtained by mass spectrometry, electrophoresis, TEM and AFM microscopy techniques, bacterial transformation, and real-time PCR provide evidence that ssDNA and plasmid DNA (pDNA) efficiently form complexes with nNi. AFM data suggest that the complexes are necklace-type structures, in which nanoparticles are randomly distributed along the DNA chains, rather than highly entangled clot-type structures. After desorption, observed DNA characteristics in bioanalytical and biological systems remain unchanged. Only supercoiled pDNA was nicked, but remained, as well as a plasmid-nNi complex, active in expression vector assays. These results are very important for creation of new methods of DNA immobilization and controlled manipulation.

  8. Small incision lenticule extraction (SMILE) and femtosecond laser LASIK: comparison of corneal wound healing and inflammation

    Science.gov (United States)

    Dong, Zixian; Zhou, Xingtao; Wu, Jihong; Zhang, Zhehuan; Li, Tao; Zhou, Zimei; Zhang, Shenghai; Li, Gang

    2014-01-01

    Aim To evaluate and compare early corneal wound healing and inflammatory responses after small incision lenticule extraction (SMILE) versus femtosecond laser laser in situ keratomileusis (LASIK). Methods Thirty-six eyes of 36 rabbits underwent SMILE, while another 36 eyes of 36 rabbits were treated with femtosecond laser LASIK. All the eyes were subjected to the same refractive correction of −6.00 DS/−1.00 DC. Twelve eyes that had no surgery were included for control. After euthanisation, corneal tissue sections were evaluated with terminal deoxyribonucleotidyl transferase-mediated dUTP-digoxigenin nick-end labelling (TUNEL) assay to detect apoptosis at postoperative 4 and 24 h, immunocytochemistry for Ki67 to detect keratocyte proliferation at postoperative day 3, week 1 and month 1, and immunocytochemistry for CD11b to detect inflammation at postoperative day 1, day 3 and week 1, respectively. Results No adverse effects were noted after SMILE or LASIK. Corneal healing postoperatively was uneventful in all cases. There were significantly fewer TUNEL-positive corneal stromal cells after the SMILE procedure at 4 and 24 h postoperatively (pLASIK procedure. In addition, immunocytochemistry showed significantly fewer Ki67-positive cells in the SMILE group than those in the femtosecond laser LASIK group at day 3 and week 1 postoperatively (pLASIK. PMID:24227802

  9. Spotlight on medicinal chemistry education.

    Science.gov (United States)

    Pitman, Simone; Xu, Yao-Zhong; Taylor, Peter; Turner, Nicholas; Coaker, Hannah; Crews, Kasumi

    2014-05-01

    The field of medicinal chemistry is constantly evolving and it is important for medicinal chemists to develop the skills and knowledge required to succeed and contribute to the advancement of the field. Future Medicinal Chemistry spoke with Simone Pitman (SP), Yao-Zhong Xu (YX), Peter Taylor (PT) and Nick Turner (NT) from The Open University (OU), which offers an MSc in Medicinal Chemistry. In the interview, they discuss the MSc course content, online teaching, the future of medicinal chemistry education and The OU's work towards promoting widening participation. SP is a Qualifications Manager in the Science Faculty at The OU. She joined The OU in 1993 and since 1998 has been involved in the Postgraduate Medicinal Chemistry provision at The OU. YX is a Senior Lecturer in Bioorganic Chemistry at The OU. He has been with The OU from 2001, teaching undergraduate courses of all years and chairing the master's course on medicinal chemistry. PT is a Professor of Organic Chemistry at The OU and has been involved with the production and presentation of The OU courses in Science and across the university for over 30 years, including medicinal chemistry modules at postgraduate level. NT is a Lecturer in Analytical Science at The OU since 2009 and has been involved in the production of analytical sciences courses, as well as contributing to the presentation of a number of science courses including medicinal chemistry.

  10. Quantitative and qualitative retinal microvascular characteristics and blood pressure.

    Science.gov (United States)

    Cheung, Carol Y; Tay, Wan T; Mitchell, Paul; Wang, Jie J; Hsu, Wynne; Lee, Mong L; Lau, Qiangfeng P; Zhu, Ai L; Klein, Ronald; Saw, Seang M; Wong, Tien Y

    2011-07-01

    The present study examined the effects of blood pressure on a spectrum of quantitative and qualitative retinal microvascular signs. Retinal photographs from the Singapore Malay Eye Study, a population-based cross-sectional study of 3280 (78.7% response) persons aged 40-80 years, were analyzed. Quantitative changes in the retinal vasculature (branching angle, vascular tortuosity, fractal dimension, and vascular caliber) were measured using a semi-automated computer-based program. Qualitative signs, including focal arteriolar narrowing (FAN), arteriovenous nicking (AVN), opacification of the arteriolar wall (OAW), and retinopathy (e.g., microaneurysms, retinal hemorrhages), were assessed from photographs by trained technicians. After excluding persons with diabetes and ungradable photographs, 1913 persons provided data for this analysis. In multivariable linear regression models controlling for age, sex, BMI, use of antihypertensive medication, and other factors, retinal arteriolar branching asymmetry ratio, arteriolar tortuosity, venular tortuosity, fractal dimension, arteriolar caliber, venular caliber, FAN, AVN, and retinopathy were independently associated with mean arterial blood pressure. In contrast, arteriolar/venular branching angle, venular branching asymmetry ratio and OAW were not related to blood pressure. Retinal arteriolar caliber (sβ = -0.277) and FAN (sβ = 0.170) had the strongest associations with mean arterial blood pressure, and higher blood pressure levels were associated with increasing number of both quantitative and qualitative retinal vascular signs (P trend qualitative retinal vascular signs, with the number of signs increasing with higher blood pressure levels.

  11. Player Perspectives: What It Means to Be a Gamer

    Directory of Open Access Journals (Sweden)

    Aditya Deshbandhu

    2016-12-01

    Full Text Available The term gamer has been applied in various, often simplistic ways to anyone who plays video games as a preferred leisure activity. Being heavily value-laden both socially and culturally, the term has spawned fiery debates on a variety of issues ranging from perpetuation of stereotypes to gender inequality. This paper seeks to understand the gamer tag by charting and critically analysing the gaming journeys of three video game players in an everyday context. The paper uses an ethnographic approach, based on in-depth interviews with and observation of the gaming activity of three individuals over a year. Drawing from Nick Couldry’s Media as Practice approach the gaming practices of the players were examined in relation to how they themselves reflect upon their experience and its various components, and the ways in which they construct and express their gamer/gaming identity. The emergent themes from the analysis have been used to build a tentative framework that could enable a more holistic understanding of the gamer within the gaming world and more generally in popular culture.

  12. Missed cleavage opportunities by FEN1 lead to Okazaki fragment maturation via the long-flap pathway

    KAUST Repository

    Zaher, Manal S.

    2018-01-27

    RNA-DNA hybrid primers synthesized by low fidelity DNA polymerase α to initiate eukaryotic lagging strand synthesis must be removed efficiently during Okazaki fragment (OF) maturation to complete DNA replication. In this process, each OF primer is displaced and the resulting 5\\'-single-stranded flap is cleaved by structure-specific 5\\'-nucleases, mainly Flap Endonuclease 1 (FEN1), to generate a ligatable nick. At least two models have been proposed to describe primer removal, namely short- and long-flap pathways that involve FEN1 or FEN1 along with Replication Protein A (RPA) and Dna2 helicase/nuclease, respectively. We addressed the question of pathway choice by studying the kinetic mechanism of FEN1 action on short- and long-flap DNA substrates. Using single molecule FRET and rapid quench-flow bulk cleavage assays, we showed that unlike short-flap substrates, which are bound, bent and cleaved within the first encounter between FEN1 and DNA, long-flap substrates can escape cleavage even after DNA binding and bending. Notably, FEN1 can access both substrates in the presence of RPA, but bending and cleavage of long-flap DNA is specifically inhibited. We propose that FEN1 attempts to process both short and long flaps, but occasional missed cleavage of the latter allows RPA binding and triggers the long-flap OF maturation pathway.

  13. Distribution of Ds-like sequences in genomes of cereals

    International Nuclear Information System (INIS)

    Vershinin, A.V.; Salina, E.A.; Shumnii, V.K.; Svitashev, S.K.

    1986-01-01

    It has been suggested that insertions of Ds-elements may alter the effectiveness of transcription or translation of the genetic loci and the normal processing of introns and exons, and that they may impair coding frames, etc. The object of the present study was to determine the frequency of occurence of DNA sequences similar to the Ds-controlling elements of mazie (Ds-like sequences) among other representatives of cereals. The conservative feature of the primary structure of transposons from different eukaryotic species served as a basis in this investigation. By means of the ''nick-translation'' reaction with the aid of DNA-polymerase I (alpha- 32 P) dCTP or TTP was introduced into the Ds-element. The specific radioactivity of the preparations obtained was 5 x 10 7 to 1 x 10 8 cpm/gamma. From the results obtained, it is suggested that the genomes of cereals examined contain a collection of Ds-like sequences. The Ds-element may have a significant effect on gene expression in the presence of Ac-like or other sequences, which undergo transposition

  14. Homocysteine Aggravates Cortical Neural Cell Injury through Neuronal Autophagy Overactivation following Rat Cerebral Ischemia-Reperfusion

    Directory of Open Access Journals (Sweden)

    Yaqian Zhao

    2016-07-01

    Full Text Available Elevated homocysteine (Hcy levels have been reported to be involved in neurotoxicity after ischemic stroke. However, the underlying mechanisms remain incompletely understood to date. In the current study, we hypothesized that neuronal autophagy activation may be involved in the toxic effect of Hcy on cortical neurons following cerebral ischemia. Brain cell injury was determined by hematoxylin-eosin (HE staining and TdT-mediated dUTP Nick-End Labeling (TUNEL staining. The level and localization of autophagy were detected by transmission electron microscopy, western blot and immunofluorescence double labeling. The oxidative DNA damage was revealed by immunofluorescence of 8-Hydroxy-2′-deoxyguanosine (8-OHdG. Hcy treatment aggravated neuronal cell death, significantly increased the formation of autophagosomes and the expression of LC3B and Beclin-1 in the brain cortex after middle cerebral artery occlusion-reperfusion (MCAO. Immunofluorescence analysis of LC3B and Beclin-1 distribution indicated that their expression occurred mainly in neurons (NeuN-positive and hardly in astrocytes (GFAP-positive. 8-OHdG expression was also increased in the ischemic cortex of Hcy-treated animals. Conversely, LC3B and Beclin-1 overexpression and autophagosome accumulation caused by Hcy were partially blocked by the autophagy inhibitor 3-methyladenine (3-MA. Hcy administration enhanced neuronal autophagy, which contributes to cell death following cerebral ischemia. The oxidative damage-mediated autophagy may be a molecular mechanism underlying neuronal cell toxicity of elevated Hcy level.

  15. Replication of kinetoplast minicircle DNA

    International Nuclear Information System (INIS)

    Sheline, C.T.

    1989-01-01

    These studies describe the isolation and characterization of early minicircle replication intermediates from Crithidia fasciculata, and Leishmania tarentolae, the mitochondrial localization of a type II topoisomerase (TIImt) in C. fasciculata, and the implication of the aforementioned TIImt in minicircle replication in L. tarentolae. Early minicircle replication intermediates from C. fasciculata were identified and characterized using isolated kinetoplasts to incorporate radiolabeled nucleotides into its DNA. The pulse-label in an apparent theta-type intermediate chase into two daughter molecules. A uniquely gapped, ribonucleotide primed, knotted molecule represents the leading strand in the model proposed, and a highly gapped molecule represents the lagging strand. This theta intermediate is repaired in vitro to a doubly nicked catenated dimer which was shown to result from the replication of a single parental molecule. Very similar intermediates were found in the heterogeneous population of minicircles of L. tarentolae. The sites of the Leishmania specific discontinuities were mapped and shown to lie within the universally conserved sequence blocks in identical positions as compared to C. fasciculata and Trypanosoma equiperdum

  16. Islet-1 is required for ventral neuron survival in Xenopus

    International Nuclear Information System (INIS)

    Shi, Yu; Zhao, Shuhua; Li, Jiejing; Mao, Bingyu

    2009-01-01

    Islet-1 is a LIM domain transcription factor involved in several processes of embryonic development. Xenopus Islet-1 (Xisl-1) has been shown to be crucial for proper heart development. Here we show that Xisl-1 and Xisl-2 are differentially expressed in the nervous system in Xenopus embryos. Knock-down of Xisl-1 by specific morpholino leads to severe developmental defects, including eye and heart failure. Staining with the neuronal markers N-tubulin and Xisl-1 itself reveals that the motor neurons and a group of ventral interneurons are lost in the Xisl-1 morphants. Terminal dUTP nick-end labeling (TUNEL) analysis shows that Xisl-1 morpholino injection induces extensive apoptosis in the ventral neural plate, which can be largely inhibited by the apoptosis inhibitor M50054. We also find that over-expression of Xisl-1 is able to promote cell proliferation and induce Xstat3 expression in the injected side, suggesting a potential role for Xisl-1 in the regulation of cell proliferation in co-operation with the Jak-Stat pathway.

  17. Potential protection of green tea polyphenols against 1800 MHz electromagnetic radiation-induced injury on rat cortical neurons.

    Science.gov (United States)

    Liu, Mei-Li; Wen, Jian-Qiang; Fan, Yu-Bo

    2011-10-01

    Radiofrequency electromagnetic fields (EMF) are harmful to public health, but the certain anti-irradiation mechanism is not clear yet. The present study was performed to investigate the possible protective effects of green tea polyphenols against electromagnetic radiation-induced injury in the cultured rat cortical neurons. In this study, green tea polyphenols were used in the cultured cortical neurons exposed to 1800 MHz EMFs by the mobile phone. We found that the mobile phone irradiation for 24 h induced marked neuronal cell death in the MTT (3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyl-tetrazolium bromide) and TUNEL (TdT mediated biotin-dUTP nicked-end labeling) assay, and protective effects of green tea polyphenols on the injured cortical neurons were demonstrated by testing the content of Bcl-2 Assaciated X protein (Bax) in the immunoprecipitation assay and Western blot assay. In our study results, the mobile phone irradiation-induced increases in the content of active Bax were inhibited significantly by green tea polyphenols, while the contents of total Bax had no marked changes after the treatment of green tea polyphenols. Our results suggested a neuroprotective effect of green tea polyphenols against the mobile phone irradiation-induced injury on the cultured rat cortical neurons.

  18. Punishment of Minor Female Genital Ritual Procedures: Is the Perfect the Enemy of the Good?

    Science.gov (United States)

    Jacobs, Allan J; Arora, Kavita Shah

    2017-08-01

    Female genital alteration (FGA) is any cutting, removal or destruction of any part of the external female genitalia. Various FGA practices are common throughout the world. While most frequent in Africa and Asia, transglobal migration has brought ritual FGA to Western nations. All forms of FGA are generally considered undesirable for medical and ethical reasons when performed on minors. One ritual FGA procedure is the vulvar nick (VN). This is a small laceration to the vulva that does not cause morphological changes. Besides being performed as a primary ritual procedure it has been proposed as a substitute for more extensive forms of FGA. Measures advocated or taken to reduce the burden of FGA can be punitive or non-punitive. Even if it is unethical to perform VN, we argue that it also is unethical to attempt to suppress it through punishment. First, punishment of VN is likely to cause more harm than good overall, even to those ostensibly being protected. Second, punishment is likely to exceed legitimate retributive ends. We do not argue in favor of performing VN. Rather, we argue that non-punitive strategies such as education and harm reduction should be employed. © 2016 John Wiley & Sons Ltd.

  19. Relationship between bcl-2, bax, beclin-1, and cathepsin-D proteins during postovulatory follicular regression in fish ovary.

    Science.gov (United States)

    Morais, Roberto D V S; Thomé, Ralph G; Santos, Hélio B; Bazzoli, Nilo; Rizzo, Elizete

    2016-04-01

    In fish ovaries, postovulatory follicles (POFs) are key biomarkers of breeding and provide an interesting model for studying the relationship between autophagy and apoptosis. In this study, we investigated the immunohistochemical expression of autophagic and apoptotic proteins to improve the knowledge on the mechanisms regulating ovarian remodeling after spawning. Females from three neotropical fish species kept in captivity were submitted to hormonal induction. After ova stripping, ovarian sections were sampled daily until 5 days postspawning (dps). Similar events of POF regression were detected by histology, terminal transferase-mediated dUTP nick-end labeling (TUNEL), and electron microscopy in the three species: follicular cells hypertrophy, progressive disintegration of the basement membrane, gradual closing of the follicular lumen, theca thickening, and formation of large autophagic vacuoles preceding apoptosis of the follicular cells. Autophagic and apoptotic proteins were assessed by immunohistochemistry. Morphometric analysis of the immunolabeling revealed a more intense reaction for bcl-2 and beclin-1 (BECN1) in POFs at 0 to 1 dps and for bax at 2 to 3 dps (P family, BECN1, and cathepsin-D can be involved in the regulation of ovarian remodeling in teleost fish. Copyright © 2016 Elsevier Inc. All rights reserved.

  20. Apoptosis induction of human endometriotic epithelial and stromal cells by noscapine

    Directory of Open Access Journals (Sweden)

    Mohammad Rasoul Khazaei

    2016-09-01

    Full Text Available Objective(s: Endometriosis is a complex gynecologic disease with unknown etiology. Noscapine has been introduced as a cancer cell suppressor. Endometriosis was considered as a cancer like disorder, The aim of present study was to investigate noscapine apoptotic effect on human endometriotic epithelial and stromal cells in vitro. Materials and Methods:In this in vitro study, endometrial biopsies from endometriosis patients (n=9 were prepared and digested by an enzymatic method (collagenase I, 2 mg/ml. Stromal and epithelial cells were separated by sequential filtration through a cell strainer and ficoll layering. The cells of each sample were divided into five groups: control (0, 10, 25, 50 and 100 micromole/liter (µM concentration of noscapine and were cultured for three different periods of times; 24, 48 and 72 hr. Cell viability was assessed by colorimetric assay. Nitric oxide (NO concentration was measured by Griess reagent. Cell death was analyzed by Acridine Orange (AO–Ethidium Bromide (EB double staining and Terminal deoxynucleotidyl transferase (TdT dUTP Nick-End Labeling (TUNEL assay. Data were analyzed by one-way ANOVA. Results: Viability of endometrial epithelial and stromal cells significantly decreased in 10, 25, 50 and 100 µM noscapine concentration in 24, 48, 72 hr (P